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Cannabinoid receptor agonists are mitochondrial inhibitors: A unified hypothesis of how cannabinoids modulate mitochondrial function and induce cell death
Abstract
Time-lapse microscopy of human lung cancer (H460) cells showed that the endogenous cannabinoid anandamide (AEA), the phyto-cannabinoid Delta-9-tetrahydrocannabinol (THC) and a synthetic cannabinoid HU 210 all caused morphological changes characteristic of apoptosis. Janus green assays of H460 cell viability showed that AEA and THC caused significant increases in OD 595 nm at lower concentrations (10-50 microM) and significant decreases at 100 microM, whilst HU 210 caused significant decreases at all concentrations. In rat heart mitochondria, all three ligands caused significant decreases in oxygen consumption and mitochondrial membrane potential. THC and HU 210 caused significant increases in mitochondrial hydrogen peroxide production, whereas AEA was without significant effect. All three ligands induced biphasic changes in either mitochondrial complex I activity and/or mitochondrial complex II-III activity. These data demonstrate that AEA, THC, and HU 210 are all able to cause changes in integrated mitochondrial function, directly, in the absence of cannabinoid receptors.
... Athanasiou and colleagues were among the first to characterize the impacts of AEA in vitro, observing that 100 mM AEA induced an apoptotic phenotype in human lung cancer cells (H460 cell line). Specifically, they noted an increased appearance of rounding and unadhered cells combined with increased cytoplasmic granularity following AEA treatment [139]. Like other cannabinoid receptor agonists, such as THC, they demonstrated that AEA treatment in isolated rat mitochondria led to decreased mitochondrial membrane potential upon quantifying Rhodamine 123 fluorescence using fluorimeter tracing [139]. ...
... Specifically, they noted an increased appearance of rounding and unadhered cells combined with increased cytoplasmic granularity following AEA treatment [139]. Like other cannabinoid receptor agonists, such as THC, they demonstrated that AEA treatment in isolated rat mitochondria led to decreased mitochondrial membrane potential upon quantifying Rhodamine 123 fluorescence using fluorimeter tracing [139]. Most notably, it was discovered that incremental increases in AEA concentration resulted in slightly enhanced complex II-III activity at low micromolar doses, which was short-lived as complex II-III activity drastically decreased while approaching the higher micromolar range [139]. ...
... Like other cannabinoid receptor agonists, such as THC, they demonstrated that AEA treatment in isolated rat mitochondria led to decreased mitochondrial membrane potential upon quantifying Rhodamine 123 fluorescence using fluorimeter tracing [139]. Most notably, it was discovered that incremental increases in AEA concentration resulted in slightly enhanced complex II-III activity at low micromolar doses, which was short-lived as complex II-III activity drastically decreased while approaching the higher micromolar range [139]. This evident disruption in ETC activity may be attributed to mtCB1 activation, as Bénard et al. have previously shown that mtCB1 signaling results in decreased PKA-dependent phosphorylation of NDUFS2, a crucial complex I subunit, thus decreasing mitochondrial respiration [90]. ...
The endocannabinoid system (ECS) governs and coordinates several physiological processes through an integrated signaling network, which is responsible for inducing appropriate intracellular metabolic signaling cascades in response to (endo)cannabinoid stimulation. This intricate cellular system ensures the proper functioning of the immune, reproductive, and nervous systems and is involved in the regulation of appetite, memory, metabolism, and development. Cannabinoid receptors have been observed on both cellular and mitochondrial membranes in several tissues and are stimulated by various classes of cannabinoids, rendering the ECS highly versatile. In the context of growth and development, emerging evidence suggests a crucial role for the ECS in cellular growth and differentiation. Indeed, cannabinoids have the potential to disrupt key energy-sensing metabolic signaling pathways requiring mitochondrial-ER crosstalk, whose functioning is essential for successful cellular growth and differentiation. This review aims to explore the extent of cannabinoid-induced cellular dysregulation and its implications for cellular differentiation.
... GPCRs are classically seen as presynaptic plasma membrane proteins in order to convert extracellular signals into intracellular responses (Freund and Hajos, 2003). Until recently CB1-dependent effects have been exclusively attributed to plasma membrane CB1 (pmCB1) and its possible presence at intracellular level was disputed mainly for the aspecificity of the anti-CB1 immunolabelling (Campbell, 2001;Athanasiou et al. 2007). However, the idea that GPCRs targeted by water-soluble ligands were believed to be functional only at plasma membranes has been challenged over the years by some studies, showing that different types of GPCRs can also be located intracellularly, including endosomes, nuclei and mitochondria ( Figure 12) (Irannejad et al. 2013(Irannejad et al. , 2017Jong et al. 2014;Tsvetanova et al. 2015;Jalink and Moolenaar, 2010;Belous et al. 2004;Irannejad and von Zastrow, 2014;reviewed in Busquets Garcia et al. 2016). ...
... Already in the Seventies, different studies reported the impact of exogenous cannabinoids on mitochondrial processes, including reduction of complex I or V activities of the respiratory chain and changes in mitochondrial structure (Bartova and Birmingham, 1976;Bino et al. 1972;Chari-Bitron and Bino, 1971;Mahoney and Harris, 1972;Schurr and Livne, 1975). With the identification of CB1 receptors as typical plasma membrane GPCRs, these effects were explained and ascribed to indirect signaling of CB1 receptor-dependent activation (Campbell, 2001;Athanasiou et al. 2007) This study and subsequently also the one by Hébert-Chatelain (2016), Koch and colleagues (2015) have been demonstrated the importance of mitochondrial CB1 receptors coupled to the Gi/o protein in physiology and pathophysiology, using experiments with mice lacking CB1 receptor. ...
... Table S1. PART 2 -LINKING MITOCHONDRIAL G-PROTEIN SIGNALING TO CANNABINOIDS-INDUCED AMNESIA: A NEW MITOCHONDRIA-SPECIFIC CHEMOGENETIC STRATEGY Until recently physiological and behavioral CB1R-dependent effects had been exclusively attributed to canonical activation of plasma membrane CB1Rs (Campbell, 2001;Athanasiou et al. 2007). With the identification of intracellular mitochondrial CB1Rs in different tissues, in particular in the brain (Aquila et al, 2010;Benard et al, 2012;Hebert-Chatelain et al, 2014;Mendizabal-Zubiaga et al, 2016), this idea has been progressively changing. ...
Via modulation of neuronal activity by cannabinoid receptor type-1 (CB1), the endocannabinoid system represents a major brain modulatory system controlling memory functions. On the other hand, several reports point out a crucial role of hippocampal dopamine signaling in the regulation of memory related processes. Furthermore, recent evidence suggests that hippocampal cells expressing dopamine receptors do also posses CB1 receptors.The work presented in this Thesis aims at establishing a functional connection between CB1 receptor and dopaminoceptive signaling in the regulation of hippocampal related memory processes with particular enfasis on the cellular and sub-cellular mechanisms involved.In the first part of the thesis we observed that a mouse line lacking CB1 in dopamine receptor type- 1 cells (D1-CB1-KO) displayed impaired long-term novel object recognition memory (NOR) and, interestingly, viral-mediated re-expression of CB1 in D1-positive cells in the hippocampus of D1-CB1- KO mice reversed the NOR impairment present in these mice. Furthermore, we pointed out execessive hippocampal GABAA receptor activation and impaired in vivo long-term potentiation (LTP) in the CA3-CA1 pathway as the main cellular mechanisms for memory impairment in D1-CB1- KO. Thus, we provided functional evidence for the involvement of a small subclass of type-1 cannabinoid receptor (CB1)-expressing hippocampal interneurons in the modulation of specific hippocampal circuits in memory processes.The second part of the Thesis focused on subcellular location of CB1 activation in D1 positive cells. Indeed, besides the canonical regulation of neuronal activity by plasma membrane CB1 receptor, recent evidence suggests the involvement of mitochondrial CB1 receptor (mtCB1) in the regulation of bioenergetic processes which impacts on synaptic transmission and amnesic effects of cannabinoids. We found that mtCB1 receptors in hippocampal D1-positive neurons is not required for physiological regulation of memory formation per se but its activation is required for THC- induced memory impairment. Looking for the intracellular and intra-mitochondrial G-protein signaling involved in these processes, we developed a new chemogenetic strategy which specifically modulates the mitochondrial G-protein signaling and we observed its contribution in brain mitochondrial activity and cognitive functions. Specific chemogenetic activation of mitochondrial G- protein signaling results in increased mitochondrial respiration which in turns rescues THC-induced amnesic effect.Overall, the results of this Thesis indicate the mechanisms linking the diversity of cellular and subcellular CB1 receptors in higher brain functions, including learning and memory and provide the basis for the development of more selective and precise therapeutic strategies for cognitive disorders.
... With it came a new swarth of papers suggesting that both THC and CBD could modulate mitochondrial function. For instance, THC could disrupt mitochondrial function, inhibit respiration and could result in cell death [109,110], and it could biphasically affect complex 1 function [111], while it could also restore mitochondrial function, possibly via activation of the peroxisomal proliferating activate receptor gamma (PPARγ), in a neuroblastoma cell line [112]. In an interesting twist, Fisar and colleagues also suggested that THC had both direct and receptor-mediated effects on mitochondrial complexes, supporting some evidence that mitochondria could contain a cannabinoid receptor (CBR), as well as indicating that CBD also affected the ETC: the IC 50 in relation to respiration was 8.2 and 15.1 µM for CBD and THC, respectively [113]. ...
... Another potential possibility is suggested by the observation that VDAC1 plays a role in the conduction of ROS generated from complex 1 [210]; this would suggest that inhibiting its function might reduce excessive cytosolic ROS signalling. However, these compounds have also potentially been shown to directly modulate the ETC [111,113], which could be key in how they result in different effects in different cells by modulation of ROS from different complexes. For instance, there is some evidence of a synergistic effect of THC and CBD in inhibiting respiration by downregulating respiratory chain proteins in glioblastoma cells [123], while CBD has been shown to increase the mitochondrial complex and creatine kinase activity in rat brains [211]. ...
Before the late 1980s, ideas around how the lipophilic phytocannabinoids might be working involved membranes and bioenergetics as these disciplines were “in vogue”. However, as interest in genetics and pharmacology grew, interest in mitochondria (and membranes) waned. The discovery of the cognate receptor for tetrahydrocannabinol (THC) led to the classification of the endocannabinoid system (ECS) and the conjecture that phytocannabinoids might be “working” through this system. However, the how and the “why” they might be beneficial, especially for compounds like CBD, remains unclear. Given the centrality of membranes and mitochondria in complex organisms, and their evolutionary heritage from the beginnings of life, revisiting phytocannabinoid action in this light could be enlightening. For example, life can be described as a self-organising and replicating far from equilibrium dissipating system, which is defined by the movement of charge across a membrane. Hence the building evidence, at least in animals, that THC and CBD modulate mitochondrial function could be highly informative. In this paper, we offer a unique perspective to the question, why and how do compounds like CBD potentially work as medicines in so many different conditions? The answer, we suggest, is that they can modulate membrane fluidity in a number of ways and thus dissipation and engender homeostasis, particularly under stress. To understand this, we need to embrace origins of life theories, the role of mitochondria in plants and explanations of disease and ageing from an adaptive thermodynamic perspective, as well as quantum mechanics.
... antagonists not used [193] rats cardiac mitochondria THC, HU-210, AEA THC and HU-210 ↓oxygen consumption and ↓mitochondrial membrane potential ↑mitochondrial hydrogen peroxide production ↓mitochondrial oxygen consumption; antagonists not used [194] Wistar rats cardiac mitochondria THC up to 500 µM ↔ROS production, no mitochondrial swelling ↔membrane potential, no oxidative stress, no lipid peroxidation THC is not directly toxic in isolated cardiac mitochondria, and may even be helpful in reducing mitochondrial toxicity [195] SD rats neonatal ventricular myocytes CB13 ...
... However, the results obtained so far are contradictory. Thus, on the one hand, THC or AEA and HU-210 not only at high (100-120 or 1-20 µM) but even at low (0.1 or 0.2 µM) concentrations led to a decrease in oxygen consumption in bovine [193], rat [194] and mouse [191,192] cardiac tissue or mitochondria. The latter was connected with a lower mitochondrial membrane potential and an enhanced mitochondrial hydrogen peroxide production ( Table 4). ...
The use of cannabis preparations has steadily increased. Although cannabis was traditionally assumed to only have mild vegetative side effects, it has become evident in recent years that severe cardiovascular complications can occur. Cannabis use has recently even been added to the risk factors for myocardial infarction. This review is dedicated to pathogenetic factors contributing to cannabis-related myocardial infarction. Tachycardia is highly important in this respect, and we provide evidence that activation of CB1 receptors in brain regions important for cardiovascular regulation and of presynaptic CB1 receptors on sympathetic and/or parasympathetic nerve fibers are involved. The prototypical factors for myocardial infarction, i.e., thrombus formation and coronary constriction, have also been considered, but there is little evidence that they play a decisive role. On the other hand, an increase in the formation of carboxyhemoglobin, impaired mitochondrial respiration, cardiotoxic reactions and tachyarrhythmias associated with the increased sympathetic tone are factors possibly intensifying myocardial infarction. A particularly important factor is that cannabis use is frequently accompanied by tobacco smoking. In conclusion, additional research is warranted to decipher the mechanisms involved, since cannabis use is being legalized increasingly and Δ9-tetrahydrocannabinol and its synthetic analogue nabilone are indicated for the treatment of various disease states.
... More recently, Δ9-THC has been reported to be utilized as an adjunct treatment for various neurological diseases including PD [4]. Since in PD, mitochondrial impairment plays a crucial role in cell degeneration, the use of phytocannabinoids is not quite obvious, as studies demonstrated mitochondrial toxicity of phytocannabinoids leading to mitochondrial dysfunction and an increase of oxidative stress [7,8]. But in a PD cell culture model, phytocannabinoids, especially Δ9-THC, showed antioxidant properties against MPP + and rotenone. ...
... The authors reported that exposure of human SH-SY5Y cells to 250 μM H 2 O 2 for 24 h led to 50 % cell cytotoxicity [31]. Athanasiou and colleagues [7] could show that production of H 2 O 2 was significantly triggered by μM range concentrations of THC. The same effects were found by Wolff and her colleagues [8]. ...
Delta-9-Tetrahydrocannabinol and other phytocannabinoids have been previously demonstrated to possess neuroprotective effects in murine mesencephalic cell culture models of Parkinson’s disease, in which increased levels of superoxide radicals led to the loss of dopaminergic neurons. In these models, delta-9-tetrahydrocannabinol did not scavenge these radicals but displayed antioxidative capacity by increasing glutathione levels. Based on these findings, in the present study, we investigated whether the neuroprotective effect of delta-9-tetrahydrocannabinol can also be detected in FeSO4- and H2O2-stressed cells. Mesencephalic cultures were concomitantly treated with FeSO4 (350 μM) or H2O2 (150 μM) and delta-9-tetrahydrocannabinol (0.01, 0.1, 1, 10 μM) on the 12th days in vitro for 48 h. On the 14th DIV, dopaminergic neurons were stained immunocytochemically by tyrosine hydroxylase, and fluorescently using crystal violet, Hoechst 33342, and JC-1. FeSO4 and H2O2 significantly reduced the number of dopaminergic neurons by 33 and 36%, respectively, and adversely affected the morphology of surviving neurons. Moreover, FeSO4, but not H2O2, significantly decreased the fluorescence intensity of crystal violet and Hoechst 33342, and reduced the red/green ratio of JC-1. Co-treatment with delta-9-tetrahydrocannabinol at the concentrations 0.01 and 0.1 μM significantly rescued dopaminergic neurons in FeSO4 and H2O2-treated cultures by 16 and 30%, respectively. delta-9-Tetrahydrocannabinol treatment also led to a higher fluorescence intensity of crystal violet and Hoechst 33342, and increased the red/green fluorescence ratio of JC-1 when concomitantly administered with FeSO4 but not H2O2. To conclude, delta-9-tetrahydrocannabinol rescues dopaminergic neurons against FeSO4- and H2O2-induced neurotoxicity. Using fluorescence dyes, this effect seems to be mediated partially by restoring mitochondrial integrity and decreasing cell death, particularly in FeSO4-treated cultures.
... CB1 receptor is present at membrane of neuronal mitochondria, and get activated by exogenous cannabinoid or endocannabinoid which decreases cyclic AMP concentration, protein kinase A activity, enzymatic activity and respiration in neuronal mitochondria (Bénard et al., 2012). Thus, activation of mitochondrial CB1 receptor reduces oxidative stress, ROS production and oxidative phosphorylation (Athanasiou et al., 2007). Cannabinoids plays protective role against mitochondrial damage during oxidative stress and also decrease Ca2+ induced cytochrome c release from mitochondria, further preventing neuronal cell from apoptosis (Velez-Pardo et al., 2010;Catanzaro et al., 2009). ...
... Moreover, CB1 receptor are present at membrane of neuronal mitochondria, and get activated by exogenous cannabinoid or endocannabinoid which decreases cyclic AMP concentration, protein kinase A activity, enzymatic activity and respiration in neuronal mitochondria (Bénard et al., 2012). Thus, activation of mitochondrial CB1 receptor reduces oxidative stress, ROS production and oxidative phosphorylation (Athanasiou et al., 2007). Cannabinoids plays protective role against mitochondrial damage during oxidative stress and also decrease Ca2+ induced cytochrome c release from mitochondria, further preventing neuronal cell from apoptosis (Velez-Pardo et al., 2010;Catanzaro et al., 2009). ...
Alzheimer's disease is a neurodegenerative disease characterized by progressive decline of cognitive function in combination with neuronal death. Current approved treatment target single dysregulated pathway instead of multiple mechanism, resulting in lack of efficacy in slowing down disease progression. The proclivity of endocannabinoid system to exert neuroprotective action and mitigate symptoms of neurodegeneration condition has received substantial interest. Growing evidence suggest the endocannabinoids (eCB) system, viz. anadamide (AEA) and arachidonoyl glycerol (2-AG), as potential therapeutic targets with the ability to modify Alzheimer's pathology by targeting the inflammatory, neurodegenerative and cognitive aspects of the disease. In order to modulate endocannabinoid system, number of agents have been reported amongst which are inhibitors of the monoacylglycerol (MAGL) and fatty acid amide hydrolase (FAAH), the enzymes that hydrolyses 2-AG and AEA respectively. However, little is known regarding the exact mechanistic signalling and their effects on pathophysiology and cognitive decline associated with Alzheimer's disease. Both MAGL and FAAH inhibitors possess fascinating properties that may offer a multi-faceted approach for the treatment of Alzheimer's disease such as potential to protect neurons from deleterious effect of amyloid-β, reducing phosphorylation of tau, reducing amyloid-β induced oxidative stress, stimulating neurotrophin to support brain intrinsic repair mechanism etc. Based on empirical evidence, MAGL and FAAH inhibitors might have potential for therapeutic efficacy against cognitive impairment associated with Alzheimer's disease. The aim of this review is to summarize the experimental studies demonstrating the polyvalent properties of MAGL or FAAH inhibitor compounds for the treatment of Alzheimer's disease, and also effect of these on learning and types of memories, which together encourage to study these compounds over other therapeutics targets. Further research in this direction would enhance the molecular mechanisms and development of applicable interventions for the treatment of Alzheimer's disease, which nevertheless stay as the primary unmet need.
... Additionally, maternal undernutrition in rats induces impaired placental mitochondrial function and results in fetal and placental growth restriction 33 . Recently, associations between mitochondrial dysfunction and THC exposure have been suggested 34 . THC treatment in human lung cancer cells (H460) was shown to reduce mitochondrial complex I and complex II-III activities, reduce mitochondrial membrane potential, and induce oxidative stress and apoptosis 34 . ...
... Recently, associations between mitochondrial dysfunction and THC exposure have been suggested 34 . THC treatment in human lung cancer cells (H460) was shown to reduce mitochondrial complex I and complex II-III activities, reduce mitochondrial membrane potential, and induce oxidative stress and apoptosis 34 . ...
Prenatal cannabis use is a significant problem and poses important health risks for the developing fetus. The molecular mechanisms underlying these changes are not fully elucidated but are thought to be attributed to delta-9-tetrahydrocannabinol (THC), the main bioactive constituent of cannabis. It has been reported that THC may target the mitochondria in several tissue types, including placental tissue and trophoblast cell lines, and alter their function. In the present study, in response to 48-h THC treatment of the human extravillous trophoblast cell line HTR8/SVneo, we demonstrate that cell proliferation and invasion are significantly reduced. We further demonstrate THC-treatment elevated levels of cellular reactive oxygen species and markers of lipid damage. This was accompanied by evidence of increased mitochondrial fission. We also observed increased expression of cellular stress markers, HSP70 and HSP60, following exposure to THC. These effects were coincident with reduced mitochondrial respiratory function and a decrease in mitochondrial membrane potential. Taken together, our results suggest that THC can induce mitochondrial dysfunction and reduce trophoblast invasion; outcomes that have been previously linked to poor placentation. We also demonstrate that these changes in HTR8/SVneo biology may be variably mediated by cannabinoid receptors CB1 and CB2.
... For instance, quercetin can modulate complex-1 of the ETC at low concentrations (<10 μM), reducing mitochondrial hydrogen peroxide production and inhibiting apoptosis (Lagoa, Graziani, Lopez-Sanchez, Garcia-Martinez, & Gutierrez-Merino, 2011). Similarly, berberine can also inhibit complex 1 (Turner et al., 2008), as can CBD, THC and resveratrol (Athanasiou et al., 2007;Fisar et al., 2014;Zini, Morin, Bertelli, Bertelli, & Tillement, 1999). Resveratrol, EGCG, curcumin and quercetin have also been shown to have effects on F0F1-ATPase/ATP synthase (Zheng & Ramirez, 2000). ...
... of the mode of action of many plant compounds, at least in animals, seems to involve upregulation of mitochondrial function as part of an anti-oxidant mechanism(Stevenson, 2012), as well as displaying ROS-scavenging independent actions (Sandoval-Acuna, Ferreira, & Speisky, 2014). Indeed, several well-known medicinal plant compounds can directly modulate mitochondrial function, including epigallocatechin gallate (EGCG), resveratrol(Battaglia, Salvi, & Toninello, 2005;Demos, Woolwine, Wilson, & McMillan, 1975;Oliveira, Nabavi, Daglia, Rastrelli, & Nabavi, 2016;Pereira et al., 2007;Ravanel, Tissut, & Douce, 1982;Usta et al., 2009;van Ginkel et al., 2007;Xie, Bezard, & Zhao, 2005), tetrahydrocannabinol (THC)(Athanasiou et al., 2007;Bartova & Birmingham, 1976;Fisar, Singh, & Hroudova, 2014;Mahoney & Harris, 1972) and CBD(Fisar et al., 2014;Mato, Victoria Sanchez-Gomez, & Matute, 2010;Olivas-Aguirre et al., 2019;Rimmerman et al., 2013;Ryan, Drysdale, Lafourcade, Pertwee, & Platt, 2009), as well as salicylic acid(de Souza et al., 2011;Gordon et al., 2002;Norman et al., 2004;Nulton-Persson, Szweda, & Sadek, 2004;Vlot, Dempsey, & Klessig, 2009) and curcumin ...
Medicine has utilised plant‐based treatments for millennia, but precisely how they work is unclear. One approach is to use a thermodynamic viewpoint that life arose by dissipating geothermal and/or solar potential. Hence, the ability to dissipate energy to maintain homeostasis is a fundamental principle in all life, which can be viewed as an accretion system where layers of complexity have built upon core abiotic molecules. Many of these compounds are chromophoric and are now involved in multiple pathways. Plants have further evolved a plethora of chromophoric compounds that can not only act as sunscreens and redox modifiers, but also have now become integrated into a generalised stress adaptive system. This could be an extension of the dissipative process. In animals, many of these compounds are hormetic, modulating mitochondria and calcium signalling. They can also display anti‐pathogen effects. They could therefore modulate bioenergetics across all life due to the conserved electron transport chain and proton gradient. In this review paper, we focus on well‐described medicinal compounds, such as salicylic acid and cannabidiol and suggest, at least in animals, their activity reflects their evolved function in plants in relation to stress adaptation, which itself evolved to maintain dissipative homeostasis.
... Mitochondrial dysfunction is the primary source of free radicals responsible for oxidative stress [52]. Cannabis has been described to induce mitochondrial dysfunction [53]. THC induced a significant production of ROS. ...
The abuse of Cannabis is a widespread issue in the Asir region. It has a lot of legal and occupational repercussions. The purpose of this study was to evaluate the health status of cannabis addicts at admission and after treatment using body mass index, glycemic status, liver function, renal function, and oxidative stress. A cross-sectional study was conducted with 120 participants. The study was conducted at Al Amal Hospital for Mental Health in Asir region of Saudi Arabia, with 100 hospitalized patients receiving addiction treatment and 20 healthy volunteers. The participants were divided into two groups: group I, the control group, and group II, the cannabis addicts. The socio-demographic data were gathered. The level of cannabis in the urine and the CWAS [Cannabis Withdrawal Assessment Scale] were determined. In addition, the Body Mass Index [BMI], vital signs [temperature, heart rate, systolic blood pressure, diastolic blood pressure, and respiratory rate], serum levels of albumin, total bilirubin, direct bilirubin, AST, ALT, and ALP, urea, creatinine, Thiobarbituric acid-reactive substances [TBARS], superoxide dismutase [SOD], reduced glutathione [GSH], and catalase [CAT] were analyzed on the first day of admission and after treatment. According to the results, there was no significant change in the body mass index. The vital signs in the cannabis user group were significantly lower than the corresponding admission values. Regarding renal function tests such as urea and creatinine, we found that after treatment, the mean urea and creatinine values in the cannabis user group did not differ significantly from the corresponding admission values. However, after treatment, the mean values of fasting blood glucose levels in the cannabis user group were significantly lower than at admission. Also, the mean values of liver function tests such as albumin, total bilirubin, direct bilirubin, AST, ALT, and ALP in the cannabis user group were significantly lower than the corresponding admission values after treatment. In assessing the antioxidant system, we found that the mean values of TBARS, SOD, GSH, and CAT in the cannabis user group did not differ significantly from the corresponding admission values after treatment. The current findings have revealed that cannabis addiction harms the various body systems and has significant implications for the addict’s state of health. The values of oxidative stress biomarkers did not change in this study, but other measured parameters improved after treatment.
... However, CB1 and CB2 receptors remain the primary site of action for CBD and both are expressed on the astrocytic membrane and its mitochondria (Stella 2004;Athanasiou et al. 2007;Busquets-Garcia et al. 2018). Astrocytes are the most abundant glial cells in the brain with several key roles such as the regulation of neuro-inflammation and neuronal metabolism (Scuderi et al. 2013;Acioglu et al. 2021). ...
Cerebral metabolic abnormalities are common in neurodegenerative diseases. Previous studies have shown that mitochondrial damage alters ATP production and increases reactive oxygen species (ROS) release which may contribute to neurodegeneration. In the present study, we investigated the neuroprotective effects of cannabidiol (CBD), a non-psychoactive component derived from marijuana (Cannabis sativa L.), on astrocytic bioenergetic balance in a primary cell culture model of lipopolysaccharide (LPS)-induced neurotoxicity. Astrocytic metabolic profiling using an extracellular flux analyzer demonstrated that CBD decreases mitochondrial proton leak, increased spare respiratory capacity and coupling efficiency in LPS-stimulated astrocytes. Simultaneously, CBD increased astrocytic glycolytic capacity and glycolysis reserve in a cannabinoid receptor type 1 (CB1)-dependent manner. CBD-restored metabolic changes were correlated with a significant decrease in the pro-inflammatory cytokines tumor necrosis factor α (TNFα) and interleukin-6 (IL-6) concentration and reduction of ROS production in LPS-stimulated astrocytes. These results suggest that CBD may inhibit LPS-induced metabolic impairments and inflammation by enhancing astrocytic metabolic glycolysis versus oxidative phosphorylation through its action on CB1 receptors. The present findings suggest CBD as a potential anti-inflammatory treatment in metabolic pathologies and highlight a possible role for the cannabinoidergic system in the modulation of mitochondrial oxidative stress.
Graphical Abstract
CBD enhances mitochondrial bioenergetic profile, attenuates proinflammatory cytokines release, and ROS overproduction of astrocytes stimulated by LPS. These effects are not mediated directly by CB1 receptors, while these receptors seem to have a key role in the anti-inflammatory response of the endocannabinoid system on astrocytes, as their specific inhibition by SR141716A led to increased pro-inflammatory cytokines release and ROS production. The graphical abstract is created with BioRender.com.
... Tissue damage caused by AB-CHMINACA could be attributed to the oxidative stress caused by the direct effect on mitochondrial respiratory enzymes with the decrease in the rate of O 2 consumption and increase in the levels of hydrogen peroxide [48]. In addition to the direct toxic effect, nephrotoxicity in some cases could be due to rhabdomyolysis from convulsion and agitation [49]. ...
Background:
AB-CHMINACA is a cannabimimetic indazole derivative. In 2013, it was reported in different countries as a substance of abuse.
Purpose:
This study evaluated the subacute toxic effects of AB-CHMINACA on the liver and kidneys and measured its blood level in adult male mice.
Methods:
The histological and biochemical subacute toxic effects on the liver and kidneys were assessed after four weeks of daily intraperitoneal injections of one of the following doses: 0.3 mg/kg, 3 mg/kg, or 10 mg/kg as the highest dose in adult male albino mice. In addition, the blood concentration level of AB-CHMINACA was determined by GC-MS-MS.
Results:
The histological effects showed congestion, hemorrhage, degeneration, and cellular infiltration of the liver and kidney tissues. Considering the control groups as a reference, biochemical results indicated a significant increase in the serum AST only in the highest dose group, while the ALT and creatinine levels did not significantly change. The mean values of AB-CHMINACA blood levels were 3.05 ± 1.16, 15.08 ± 4.30, and 54.43 ± 8.70 ng/mL for the three treated groups, respectively, one hour after the last dose of intraperitoneal injection. The calibration curves were linear in the 2.5-500 ng/mL concentration range. The intra-assay precision and accuracy of the method were less than 7.0% (RSD) and ± 9.2% (Bias).
Conclusion:
This research supports the available case reports on AB-CHMINACA toxicity that it has low lethality; still, the chronic administration causes evident liver and kidney histotoxic effects even at low doses with unnoticeable clinical effects in mice.
... This observation underscores the significance of the dose-dependent effects of CBD on mitochondrial function and emphasizes the need for further functional studies underlying the mechanisms. Other endogenous and exogenous CB1 ligands induced a biphasic in vitro change in some mitochondrial complexes (Athanasiou et al., 2007). Nevertheless, a direct comparison of respiratory chain gene activity and expression levels after different CBD dosages in animal models is necessary to fully understand the effects on mitochondrial function. ...
Background: Cannabidiol (CBD), one of the main cannabinoids present in the female flowers of Cannabis sativa, has been a therapeutic alternative for a plurality of disorders. Previous investigation has already provided insights into the CBD molecular mechanism, however, there is no transcriptome data for CBD effects on hippocampal subfields. Here, we explore the transcriptomic changes in dorsal and ventral CA1 of adult mice hippocampus after 100 mg/kg of CBD administration (i.p.) for one or seven consecutive days. Methods: C57BL/6JUnib mice were divided into 4 groups treated with either vehicle or CBD for 1 or 7 days. The collected brains were sectioned and the hippocampal subregions were laser microdissected for RNA-Seq analysis. Data alignment, quantification and analysis were conducted with the STAR Aligner/DESeq2/clusterProfiler R-package pipeline. Results: We found changes in gene expression in CA1 neurons after single and multiple CBD administrations. Furthermore, the enrichment analysis of differentially expressed genes following 7 days of CBD administration indicates a widespread decrease in the expression levels of electron transport chain and ribosome biogenesis transcripts, while chromatin modifications and synapse organization transcripts were increased. Conclusion: This dataset provides a significant contribution toward advancing our comprehension of the mechanisms responsible for CBD effects on hippocampal neurons. The findings suggest that CBD prompts a significant reduction in energy metabolism genes and the protein translation machinery in CA1 neurons.
... Mitochondria are thought to be the primary organelles involved in mediating most apoptotic pathways in mammalian cells (Chalah and Khosravi-Far, 2008). AEA directly and dose-dependently facilitates mitochondrial swelling and membrane potential loss in isolated mitochondria and human lung cancer cells (Athanasiou et al., 2007;Catanzaro et al., 2009), revealing that the intracellular uptake of excess AEA directly affects mitochondria functions, triggering toward cell apoptosis (Nunn et al., 2012). EOPF further encourages AEA-induced mitochondrial membrane potential loss and activation of apoptosis initiator and executioner (caspase-9 and caspase-3/7, respectively), therefore supporting that the CB receptor-independent pathway is also involved in OP-triggered apoptosis in NG108-15 cells. ...
Organophosphate (OP) agents are continuously utilized in large amount throughout the globe for crop protection and public health, thereby creating a potential concern on human health. OP agent as an anticholinesterase also acts on the endocannabinoid (EC)-hydrolases, i.e., fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL), to reveal unexpected adverse effects including ADHD-like behaviors in adolescent male rats. The present investigation examines a hypothesis that OP compound inhibiting the EC-hydrolase(s) dysregulates the EC-signaling system, triggering apoptosis in neuronal cells. Ethyl octylphosphonofluoridate (EOPF), as an OP probe, preferably acts on FAAH over MAGL in intact NG108-15 cells. Anandamide (AEA), an endogenous FAAH substrate, is cytotoxic in a concentration-dependent manner, although 2-arachidonoylglycerol, an endogenous MAGL substrate, gives no effect in the concentrations examined here. EOPF pretreatment markedly enhances AEA-induced cytotoxicity. Interestingly, the cannabinoid receptor blocker AM251 diminishes AEA-induced cell death, whereas AM251 does not prevent the cell death in the presence of EOPF. The consistent results are displayed in apoptosis markers evaluation (caspases and mitochondrial membrane potential). Accordingly, FAAH inhibition by EOPF suppresses AEA-metabolism, and accumulated excess AEA overstimulates both the cannabinoid receptor- and mitochondria-mediated apoptotic pathways.
... The sites of action that explain the beneficial effect of EPI (i.e., mainly owing to CBD content) on mitochondrial dysfunction induced by rotenone remain to be identified in this study, although some hypotheses could be proposed. Several studies have suggested mitochondria as targets for cannabinoids, 43 as the mitochondrial CB1 (mCB1) receptor plays a relevant role in cell metabolic activity. 44 However, it is unlikely that the mCB1 receptor could mediate the neuroprotective effect of CBD on rotenone-induced neurotoxicity because the CB1 antagonist (AM251) did not block the neuroprotective effect of CBD on rotenone-induced neurotoxicity in our previous study. ...
... This phenomenon is observed both in neurons and glial cells [175,181,193]. The CB1 receptor-induced decrease in the production of cAMP is coupled to downregulation of PKA and consequently alters the functionality of complex I, II and III of the OXPHOS chain [176,181,194]. In particular, inhibition of the phosphorylation of the complex I protein NDUFS2 plays a key role in the acute effects of cannabinoids on mitochondrial mobility, synaptic depression and eventually amnesia. ...
Mitochondria are present in the pre- and post-synaptic regions, providing the energy required for the activity of these very specialized neuronal compartments. Biogenesis of synaptic mitochondria takes place in the cell body, and these organelles are then transported to the synapse by motor proteins that carry their cargo along microtubule tracks. The transport of mitochondria along neurites is a highly regulated process, being modulated by the pattern of neuronal activity and by extracellular cues that interact with surface receptors. These signals act by controlling the distribution of mitochondria and by regulating their activity. Therefore, mitochondria activity at the synapse allows the integration of different signals and the organelles are important players in the response to synaptic stimulation. Herein we review the available evidence regarding the regulation of mitochondrial dynamics by neuronal activity and by neuromodulators, and how these changes in the activity of mitochondria affect synaptic communication.
... The sites of action that explain the beneficial effect of EPI (i.e., mainly owing to CBD content) on mitochondrial dysfunction induced by rotenone remain to be identified in this study, although some hypotheses could be proposed. Several studies have suggested mitochondria as targets for cannabinoids, 43 as the mitochondrial CB1 (mCB1) receptor plays a relevant role in cell metabolic activity. 44 However, it is unlikely that the mCB1 receptor could mediate the neuroprotective effect of CBD on rotenone-induced neurotoxicity because the CB1 antagonist (AM251) did not block the neuroprotective effect of CBD on rotenone-induced neurotoxicity in our previous study. ...
Introduction: Preclinical research supports the benefits of pharmaceutical cannabis-based extracts for treating different medical conditions (e.g., epilepsy); however, their neuroprotective potential has not been widely investigated. Materials and Methods: Using primary cultures of cerebellar granule cells, we evaluated the neuroprotective activity of Epifractan (EPI), a cannabis-based medicinal extract containing a high level of cannabidiol (CBD), components like terpenoids and flavonoids, trace levels of Δ9-tetrahydrocannabinol, and the acid form of CBD. We determined the ability of EPI to counteract the rotenone-induced neurotoxicity by analyzing cell viability and morphology of neurons and astrocytes by immunocytochemical assays. The effect of EPI was compared with XALEX, a plant-derived and highly purified CBD formulation (XAL), and pure CBD crystals (CBD). Results: The results revealed that EPI induced a significant reduction in the rotenone-induced neurotoxicity in a wide range of concentrations without causing neurotoxicity per se. EPI showed a similar effect to XAL suggesting that no additive or synergistic interactions between individual substances present in EPI occurred. In contrast, CBD did show a different profile to EPI and XAL because a neurotoxic effect per se was observed at higher concentrations assayed. Medium-chain triglyceride oil used in EPI formulation could explain this difference. Conclusion: Our data support a neuroprotective effect of EPI that may provide neuroprotection in different neurodegenerative processes. The results highlight the role of CBD as the active component of EPI but also support the need for an appropriate formulation to dilute pharmaceutical cannabis-based products that could be critical to avoid neurotoxicity at very high doses.
... However, CB1 and CB2 receptors remain the primary site of action for CBD and both are expressed on the astrocytic membrane and its mitochondria (Stella, 2004, Athanasiou et al., 2007, Busquets-Garcia et al., 2018. Astrocytes are the most abundant glial cells in the brain with several key roles such as the regulation of neuro-in ammation and neuronal metabolism (Scuderi et al., 2013, Acioglu et al., 2021. ...
Cerebral metabolic abnormalities are common in neurodegenerative diseases. Previous studies have shown that mitochondrial damage alters ATP production and increases Reactive Oxygen Species (ROS) release which may contribute to neurodegeneration. In the present study, we investigated the neuroprotective effects of cannabidiol (CBD), a non-psychoactive component derived from marijuana ( Cannabis sativa L.), on astrocytic bioenergetic balance; in a primary cell culture model of lipopolysaccharide (LPS)-induced neurotoxicity. Astrocytic metabolic profiling using an extracellular flux analyzer demonstrated that CBD decreases mitochondrial proton leak, increased spare respiratory capacity and coupling efficiency in LPS-stimulated astrocytes. Simultaneously CBD increased astrocytic glycolytic capacity and glycolysis reserve in a cannabinoid receptor type 1 (CB1) dependent manner. CBD-restored metabolic changes were correlated with a significant decrease in the pro-inflammatory cytokines tumor necrosis factor (TNF) and interleukin-6 (IL-6) concentration, and a reduction of ROS production in LPS-stimulated astrocytes. These results suggest that CBD may inhibit LPS-induced metabolic impairments and inflammation by enhancing astrocytic metabolic glycolysis versus oxidative phosphorylation through its action on CB1 receptors. The present findings suggest CBD as a potential anti-inflammatory treatment in metabolic pathologies and highlight a possible role for the cannabinoidergic system in the modulation of mitochondrial oxidative stress.
... It is noteworthy that TRPV1 activation by capsaicin has been proven to increase free cytosolic calcium and improve energy metabolism by upregulating PGC1α in C2C12 myotubes, as well as in skeletal muscles [73]. Furthermore, it has been demonstrated that TRPV1 ligands decrease mitochondrial membrane potential and oxygen consumption in isolated mitochondria from rat cardiomyocytes [74]. Notably, in the present study, we found that low doses of mAEA are responsible for the increase in mitochondrial membrane potential and induction of PGC1α expression, without affecting oxygen consumption, whereas the highest dose of mAEA turns the TRPV1 action to negatively modulate mitochondrial membrane polarization and cell survival. ...
The sphingosine 1-phosphate (S1P) and endocannabinoid (ECS) systems comprehend bioactive lipids widely involved in the regulation of similar biological processes. Interactions between S1P and ECS have not been so far investigated in skeletal muscle, where both systems are active. Here, we used murine C2C12 myoblasts to investigate the effects of S1P on ECS elements by qRT-PCR, Western blotting and UHPLC-MS. In addition, the modulation of the mitochondrial membrane potential (ΔΨm), by JC-1 and Mitotracker Red CMX-Ros fluorescent dyes, as well as levels of protein controlling mitochondrial function, along with the oxygen consumption were assessed, by Western blotting and respirometry, respectively, after cell treatment with methanandamide (mAEA) and in the presence of S1P or antagonists to endocannabinoid-binding receptors. S1P induced a significant increase in TRPV1 expression both at mRNA and protein level, while it reduced the protein content of CB2. A dose-dependent effect of mAEA on ΔΨm, mediated by TRPV1, was evidenced; in particular, low doses were responsible for increased ΔΨm, whereas a high dose negatively modulated ΔΨm and cell survival. Moreover, mAEA-induced hyperpolarization was counteracted by S1P. These findings open new dimension to S1P and endocannabinoids cross-talk in skeletal muscle, identifying TRPV1 as a pivotal target.
... The effects of gestational CS exposure on foetus neurodevelopment and its long-lasting effects are still controversial and investigations into CS-related dysfunction are limited (Athanasiou et al., 2007;De Oliveira et al., 2019). Thus, the current study aims to investigate the effects of prenatal exposure to a low dosage of CS on brain structure and function using in vivo and in vitro analysis. ...
Cannabis is the most widely used illegal drug during pregnancy, however, the effects of gestational exposure to Cannabis smoke (CS) on the central nervous system development remain uncharacterised. This study investigates the effects of maternal CS inhalation on brain function in the offspring. Pregnant mice were exposed daily to 5 min of CS during gestational days (GD) 5.5–17.5. On GD 18.5 half of the dams were euthanized for foetus removal. The offspring from the remaining dams were euthanized on postnatal days (PND) 20 and 60 for evaluation. Brain volume, cortex cell number, SOX2, histone-H3, parvalbumin, NeuN, and BDNF immunoreactivity were assessed in all groups. In addition, levels of NeuN, CB1 receptor, and BDNF expression were assessed and cortical primary neurons from rats were treated with Cannabis smoke extract (CSE) for assessment of cell viability. We found that male foetuses from the CS exposed group had decreased brain volume, whereas mice at PND 60 from the exposed group presented with increased brain volume. Olfactory bulb and diencephalon volume were found lower in foetuses exposed to CS. Mice at PND 60 from the exposed group had a smaller volume in the thalamus and hypothalamus while the cerebellum presented with a greater volume. Also, there was an increase in cortical BDNF immunoreactivity in CS exposed mice at PND 60. Protein expression analysis showed an increase in pro-BDNF in foetus brains exposed to CS. Mice at PND 60 presented an increase in mature BDNF in the prefrontal cortex (PFC) in the exposed group and a higher CB1 receptor expression in the PFC. Moreover, hippocampal NeuN expression was higher in adult animals from the exposed group. Lastly, treatment of cortical primary neurons with doses of CSE resulted in decreased cell viability. These findings highlight the potential negative neurodevelopmental outcomes induced by gestational CS exposure.
... This corresponds with the available studies that express that the activation of the mitochondrial CB1 receptor reduces the oxidative stress, ROS production, and oxidative phosphorylation. 64,65 The eCB also decreases Ca 2+ -induced cytochrome c release from the mitochondria and thus prevents the apoptosis of neuronal cells. 66 Drugs such as JWH-133, a CB-2 agonist, also reduce the derivative of lipid peroxidation and increase SOD levels in transgenic mice. ...
... CB1 is also localized on skeletal muscle mitochondria where it directly controls cellular respiration and ATP production, 25 and its activation is known to impair mitochondrial function and biogenesis. [25][26][27][28][29] This process is in part mediated through the production of ceramides, 29 lipid derivatives associated with the development of lipotoxicity and insulin resistance commonly seen in sarcopenia. 5 Lastly, CB1 overactivation is also associated with skeletal muscle development and function. ...
Background
Activation of the endocannabinoid system (ECS) is associated with the development of obesity and insulin resistance, and with perturbed skeletal muscle development. Age‐related sarcopenia is a progressive and generalized skeletal muscle disorder involving an accelerated loss of muscle mass and function, with changes in skeletal muscle protein homeostasis due to lipid accumulation and anabolic resistance. Hence, both obesity and sarcopenia share a common set of pathophysiological alterations leading to skeletal muscle impairment. The aim of this study was to characterize how sarcopenia impacts the ECS and if these modifications were related to the loss of muscle mass and function associated with aging in rats.
Methods
Six‐month‐old and 24‐month‐old male rats were used to measure the contractile properties of the plantarflexors (isometric torque–frequency relationship & concentric power–velocity relationship) and to evaluate locomotor activity, motor coordination, and voluntary gait by open field, rotarod, and catwalk tests, respectively. Levels of endocannabinoids (AEA & 2‐AG) and endocannabinoid‐like molecules (OEA & PEA) were measured by LCF‐MS/MS in plasma, skeletal muscle, and adipose tissue, while the expression of genes coding for the ECS were investigated by quantitative reverse transcription PCR (RT‐qPCR).
Results
Sarcopenia in old rats was exemplified by a 49% decrease in hindlimb muscle mass (P < 0.01), which was associated with severe impairment of isometric torque, power, voluntary locomotor activity, motor coordination, and gait quality. Sarcopenia was associated with (1) increased 2‐AG (+32%, P = 0.07) and reduced PEA and OEA levels in the plasma (−25% and −40%, respectively, P < 0.01); (2) an increased content of AEA, PEA, and OEA in subcutaneous adipose tissue (P < 0.01); and (3) a four‐fold increase of 2‐AG content in the soleus (P < 0.01) and a reduced OEA content in EDL (−80%, P < 0.01). These alterations were associated with profound modifications in the expression of the ECS genes in the adipose tissue and skeletal muscle.
Conclusions
Taken together, these findings demonstrate that circulating and peripheral tissue endocannabinoid tone are altered in sarcopenia. They also demonstrate that OEA plasma levels are associated with skeletal muscle function and loss of locomotor activity in rats, suggesting OEA could be used as a circulating biomarker for sarcopenia.
... ATP provides energy essential for flagellar movements of spermatozoa, and a decrease in the energy supply due to mitochondrial injuries can impair sperm motility [67]. The highly lipophilic feature of cannabinoids grants them access to the intracellular compartment, with mitochondria being reported as targets for cannabinoids [69,70]. Rimmerman et al. [71] showed that CBD directly modulates the outer mitochondrial membrane channel in BV-2 microglial cells, as an agonist of the voltage-dependent anion channel 1 (VDAC1), related to the regulation of cell energy. ...
Cannabidiol (CBD) is a natural cannabinoid present in the Cannabis sativa plant, widely prescribed as an anticonvulsant drug, especially for pediatric use. However, its effects on male reproduction are still little investigated. Therefore, the present study assessed the effects of CBD on the spermatogenesis and sperm quality. For this, twenty-one-day-old Swiss mice received CBD for 34 consecutive days by gavage at doses of either 15 or 30 mg/kg. Chronic exposure to CBD decreased the frequency of stages VII-VIII and XII of spermatogenesis and an increase in the frequency of stage IX were noted. Furthermore, the seminiferous epithelium height reduced at stage IX and increased at stage XII in both CBD-treated groups. There was a significant rise of sperm DNA damage, while no genotoxic effects were observed in leukocytes. The activities of superoxide dismutase and catalase decreased, while malondialdehyde levels increased in the sperm of mice treated with a higher dose of CBD. Mice exposed to 30 mg/kg of CBD showed a reduction in the mobile spermatozoa percentage and in curvilinear velocity, while straight line and average path velocity decreased in both treated groups. The number of acrosome-intact spermatozoa declined in the CBD 30 group, and the number of abnormal acrosomes raised in both CBD groups. On the other hand, the weight of reproductive organs, sperm count, and hormone levels were not affected by CBD treatment. These findings show that deregulation of the endocannabinoid system by CBD can reduce sperm quality. The mechanisms responsible may be associated with disorders during spermatogenesis, especially during the final stages of nuclear remodeling and assembly of acrosome. However, changes in mitochondrial function, as well as the reduction on the antioxidant enzyme activities during epididymal transit, at least partly, may also be involved.
... Previously, it has been shown that cannabinoids can stimulate mitochondrial ROS generation in different cells, leading to the impairment of mitochondrial function and cytochrome C release [37][38][39]. In rat decidual cells, CB1 activation led to ROS generation related to increased ceramide synthesis [39]. ...
Endometriosis is characterized by the formation and development of endometrial tissues outside the uterus, based on an imbalance between proliferation and cell death, leading to the uncontrolled growth of ectopic foci. The potential target for the regulation of these processes is the endocannabinoid system, which was found to be involved in the migration, proliferation, and survival of tumor cells. In this paper, we investigated the effect of endocannabinoid-like compounds from the N-acyl dopamine (NADA) family on the viability of stromal cells from ectopic and eutopic endometrium of patients with ovarian endometriosis. N-arachidonoyldopamine, N-docosahexaenoyldopamine, and N-oleoyldopamine have been shown to have a five-times-more-selective cytotoxic effect on endometrioid stromal cells. To study the mechanisms of the toxic effect, inhibitory analysis, measurements of caspase-3/9 activity, reactive oxygen species, and the mitochondrial membrane potential were performed. It was found that NADA induced apoptosis via an intrinsic pathway through the CB1 receptor and downstream serine palmitoyltransferase, NO synthase activation, increased ROS production, and mitochondrial dysfunction. The higher selectivity of NADA for endometriotic stromal cells and the current lack of effective drug treatment can be considered positive factors for further research of these compounds as possible therapeutic agents against endometriosis.
... Histopathological changes can be explained by oxidative stress in tissues causing cell apoptosis. Athanasiou et al. (2007), studied the effect of THC, endocannabinoids and synthetic cannabinoids on mitochondrial function. The study was carried out on in vitro cells. ...
... Direct effects could also be at play during fetal development given that cannabinoids can cross the placenta and ∆9-THC has been demonstrated to inhibit cardiomyocyte growth in isolated rat cardiomyocytes [149]. Moreover, CB1 antagonists prevent Doxorubicin-(a chemotherapy medication)-induced apoptosis in embryonic heart cells [150], while CB1 agonists (AEA, ∆9-THC, and HU-210) have been demonstrated to decrease mitochondria respiration and mitochondrial membrane potential in rat heart mitochondria [151]. Overall, these data suggest that maternal exposure to ∆9-THC leads to early onset of cardiac dysfunction associated with postnatal catch-up growth. ...
Up to 20% of pregnant women ages 18–24 consume cannabis during pregnancy. Moreover, clinical studies indicate that cannabis consumption during pregnancy leads to fetal growth restriction (FGR), which is associated with an increased risk of obesity, type II diabetes (T2D), and cardiovascular disease in the offspring. This is of great concern considering that the concentration of Δ9- tetrahydrocannabinol (Δ9-THC), a major psychoactive component of cannabis, has doubled over the last decade and can readily cross the placenta and enter fetal circulation, with the potential to negatively impact fetal development via the endocannabinoid (eCB) system. Cannabis exposure in utero could also lead to FGR via placental insufficiency. In this review, we aim to examine current pre-clinical and clinical findings on the direct effects of exposure to cannabis and its constituents on fetal development as well as indirect effects, namely placental insufficiency, on postnatal metabolic diseases.
... Recent studies showed that THC is able to induce mitochondrial toxicity and oxidative stress in brain mitochondria (Wolff et al. 2015;Beiersdorf et al. 2020). Athanasiou et al., demonstrated that THC is able to cause changes in integrated mitochondrial function, directly, in the absence of cannabinoid receptors in H460 cells (a human non-small cell lung cancer line) (Athanasiou et al. 2007). In addition, Valvassori et al., both acute and chronic injection of CBD increased the activity of the mitochondrial complexes (I, II, II-III, and IV) and creatine kinase (CK) in the rat brain (Valvassori et al. 2013). ...
Mitochondria have the main roles in myocardial tissue homeostasis, through providing ATP for the vital enzymes in intermediate metabolism, contractile apparatus and maintaining ion homeostasis. Mitochondria-related cardiotoxicity results from the exposure with illicit drugs have previously reported. These illicit drugs interference with processes of normal mitochondrial homeostasis and lead to mitochondrial dysfunction and mitochondrial-related oxidative stress. Cannabis consumption has been shown to cause ventricular tachycardia, to increase the risk of myocardial infarction (MI) and potentially sudden death. Here, we investigated this hypothesis that delta-9-tetrahydrocannabinol (Delta-9-THC) as a main cannabinoid found in cannabis could directly cause mitochondrial dysfunction. Cardiac mitochondria were isolated with mechanical lysis and differential centrifugation form rat heart. The isolated cardiac mitochondria were treated with different concentrations of THC (1, 5, 10, 50, 100 and 500 µM) for 1 hour at 37 °C. Then succinate dehydrogenase (SDH) activity, mitochondrial swelling, reactive oxygen species (ROS) formation, mitochondrial membrane potential (MMP) collapse and lipid peroxidation were measured in the treated and non-treated isolated cardiac mitochondria. Our observation showed that THC did not cause a deleterious alteration in mitochondrial functions, ROS production, MMP collapse, mitochondrial swelling, oxidative stress and lipid peroxidation in used concentrations (5-100 µM), even in several tests, toxicity showed a decreasing trend. Altogether, the results of the current study showed that THC is not directly toxic in isolated cardiac mitochondria, and even may be helpful in reducing mitochondrial toxicity.
... Additionally, ∆ 9 -THC has been shown to affect mitochondrial function in several tissues, including the placenta [148,149,222,223]. Human trophoblast cells exposed to ∆ 9 -THC have diminished mitochondrial respiration and ATP-coupling due to decreased abundance of mitochondrial chain complex proteins [148], as well as increased mitochondrial fission and decreased mitochondrial membrane potential [149]. ...
Cannabis use during pregnancy has continued to rise, particularly in developed countries, as a result of the trend towards legalization and lack of consistent, evidence-based knowledge on the matter. While there is conflicting data regarding whether cannabis use during pregnancy leads to adverse outcomes such as stillbirth, preterm birth, low birthweight, or increased admission to neonatal intensive care units, investigations into long-term effects on the offspring’s health are limited. Historically, studies have focused on the neurobehavioral effects of prenatal cannabis exposure on the offspring. The effects of cannabis on other physiological aspects of the developing fetus have received less attention. Importantly, our knowledge about cannabinoid signaling in the placenta is also limited. The endocannabinoid system (ECS) is present at early stages of development and represents a potential target for exogenous cannabinoids in utero. The ECS is expressed in a broad range of tissues and influences a spectrum of cellular functions. The aim of this review is to explore the current evidence surrounding the effects of prenatal exposure to cannabinoids and the role of the ECS in the placenta and the developing fetus.
... Histopathological changes can be explained by oxidative stress in tissues causing cell apoptosis. Athanasiou et al. (2007), studied the effect of THC, endocannabinoids and synthetic cannabinoids on mitochondrial function. The study was carried out on in vitro cells. ...
Abstract
Background: Synthetic cannabinoid (SCs) substances are intended for drug addiction while they cannot be easily detected on a regular drug screen. The danger of these substances is not only being undetected, but also their health effects are not well studied and cannot be predicted. This is one of the recent major health problems that threaten populations around the world. Aim of the study: This study is an experimental study to detect the toxic effect of acute exposure to a synthetic cannabinoid substance “AB-CHMINACA’ clinically and histopathologically in different organs in adult male albino rats. Material and methods: AB-CHMINACA was tested for dissolution in different solvents to choose the best vehicle. Doses were selected according to "Guidance on dose level selection for regulatory general toxicology studies for pharmaceuticals". Animals were injected intraperitoneal and after 24 hours, animals were sacrificed and the lung, heart, and liver were examined for histopathological changes. Results: AB-CHMINACA dissolves best in organic solvents like ethanol and DMSO. The most suitable vehicle for intraperitoneal injection of animals was ethanol-saline. After injection, animals showed CNS manifestations; depression or excitation followed by depression according to the dose. Histopathological examination of the lung, heart, and liver tissues showed generalized congestion, hemorrhage, inflammatory cell infiltration and degeneration, which increased by increasing the dose. Conclusion: AB-CHMINACA has toxic histopathological effects on the lung, heart, and liver on single-dose exposure even with minimal clinical manifestations. These effects are dose-related.
Key words AB-CHMINACA, Synthetic Cannabinoids, Experimental Study, Lung, Heart, Liver
... CB1 is also localized on skeletal muscle mitochondria where it directly controls cellular respiration and ATP production, 25 and its activation is known to impair mitochondrial function and biogenesis. [25][26][27][28][29] This process is in part mediated through the production of ceramides, 29 lipid derivatives associated with the development of lipotoxicity and insulin resistance commonly seen in sarcopenia. 5 Lastly, CB1 overactivation is also associated with skeletal muscle development and function. ...
Introduction et but de l’étude
La sarcopénie est définie comme la perte de masse et de force musculaires liée à l’âge. Elle est associée à une diminution de la protéosynthèse et à une accélération de la protéolyse au niveau du muscle squelettique. Les mécanismes sous-jacents comprennent notamment une accumulation ectopique de lipides et une résistance anabolique aux effets des acides aminés et de l’insuline. Ces deux phénomènes musculaires sont également observés au cours de l’obésité, situation aussi associée à une suractivation du système endocannabinoïde. Le but de cette étude était d’évaluer les perturbations du système endocannabinoïde chez des rats âgés et leur lien putatif avec la masse musculaire squelettique et la mobilité.
Matériel et méthodes
Des rats mâles adultes Wistar âgés de 6 ou 24 mois ont été utilisés pour évaluer leur composition corporelle (EchoMRI), leur mobilité spontanée à l’aide d’un openfield et la qualité de leur marche volontaire évaluée par Catwalk™. La masse des muscles des membres postérieurs a été mesurée et les concentrations plasmatiques de certains endocannabinoïdes (AEA, 2AG) et des N-acyléthanolamines (molécules endocannabinoid-like, PEA et OEA) ont été mesurées par LCF-MS/MS. Les différences entre les groupes ont été analysées par le test t de Student sur séries non appariées et une analyse de corrélation de Pearson a été utilisée.
Résultats et analyse statistique
En comparaison des rats adultes, les rats âgés étaient caractérisés par une quantité de masse grasse plus élevée (19 ± 1 vs 14 ± 1 %, p < 0,01) et une quantité de masse maigre plus faible (72 ± 2 vs 80 ± 2 %, p < 0,01), en partie due à une réduction de 49 % de la masse musculaire des membres postérieurs (p < 0,01), caractéristiques de la sarcopénie. Cette perte de masse musculaire était associée à une diminution de la distance parcourue (2720 ± 550 vs 4804 ± 960 cm, p < 0,01), de la vitesse de marche (12,8 ± 1,8 vs 10,1 ± 0,8 cm/s, p < 0,01), et de la durée d’activité dans l’openfield (268 ± 49 vs 374 ± 50 s, p < 0,01). L’analyse Catwalk révélait d’importantes altérations de la qualité de la marche chez les rats âgés. Les concentrations plasmatiques en AEA, 2-AG, PEA et OEA sont décrites dans le Tableau 1. Une analyse de corrélations a révélé que les teneurs plasmatiques en OEA, et dans certaines mesures en PEA, étaient corrélés à la distance parcourue (r² = 0,31, p < 0,02), à la vitesse de marche (r² = 0,20, p = 0,06) et à la durée d’activité (r² = 0,27, p < 0,03) et à de nombreux paramètres de qualité de la marche (p < 0,05).
Conclusion
Ces résultats montrent que les concentrations plasmatiques en endocannabinoïdes et en N-acyléthanolamines (OEA et PEA) sont altérées chez le rat âgé sarcopénique, et que leurs concentrations plasmatiques sont corrélées à la perte de mobilité associée au vieillissement.
... At the cellular level, involvement of the ECS has been demonstrated in metabolic processes relevant to fetal development. Indeed, mitochondrial and endoplasmic reticulum (ER) stress contribute to gestational complications, such as FGR (60), and 9-THC has been shown to decreases oxygen consumption and membrane potential of rat heart mitochondria, an effect that appears to be independent of cannabinoid receptor activation (61). Similarly, in the brain, 9-THC impedes mitochondrial respiratory rate, both through CB 1 R and nonreceptor-mediated mechanisms (62). ...
Clinical reports of cannabis use prevalence during pregnancy vary widely from 3% to upwards of 35% in North America; this disparity likely owing to underestimates from self-reporting in many cases. The rise in cannabis use is mirrored by increasing global legalization and the overall perceptions of safety, even during pregnancy. These trends are further compounded by a lack of evidence-based policy and guidelines for prenatal cannabis use, which has led to inconsistent messaging by healthcare providers and medically licensed cannabis dispensaries regarding prenatal cannabis use for treatment of symptoms, such as nausea. Additionally, the use of cannabis to self-medicate depression and anxiety during pregnancy is a growing medical concern. This review aims to summarize recent findings of clinical and preclinical data on neonatal outcomes, as well as long-term physiological and neurodevelopmental outcomes of prenatal cannabis exposure. Although many of the outcomes under investigation have produced mixed results, we consider these data in light of the unique challenges facing cannabis research. In particular, the limited longitudinal clinical studies available have not previously accounted for the exponential increase in (-)-Δ9– tetrahydrocannabinol (Δ9–THC; the psychoactive compound in cannabis) concentrations found in cannabis over the past two decades. Polydrug use and the long-term effects of individual cannabis constituents [Δ9–THC vs. cannabidiol (CBD)] are also understudied, along with sex-dependent outcomes. Despite these limitations, prenatal cannabis exposure has been linked to low birth weight, and emerging evidence suggests that prenatal exposure to Δ9–THC, which crosses the placenta and impacts placental development, may have wide-ranging physiological and neurodevelopmental consequences. The long-term effects of these changes require more rigorous investigation, though early reports suggest Δ9–THC increases the risk of cognitive impairment and neuropsychiatric disease, including psychosis, depression, anxiety, and sleep disorders. In light of the current trends in the perception and use of cannabis during pregnancy, we emphasize the social and medical imperative for more rigorous investigation of the long-term effects of prenatal cannabis exposure.
... Since neurons are permanent cells, they are most susceptible to hypoxic injury as that occurs with cannabis. Cannabis causes significant decrease in oxygen consumption and significant increase in mitochondrial hydrogen peroxide production and so changes in integrated mitochondrial function and inhibitions of mitochondrial respiratory chain (Athanasiou et al., 2007). ...
... Activation of CB1 receptors on the neuronal membrane decreases firing frequency and protects against depletion of energy sources, whereas activation of mitochondrial CB1 receptors decreases mitochondria activity, thus enabling a coupling between firing activity and energy need of the neurons [59]. Moreover, CB1r agonists decrease oxygen consumption, ROS production [60], and oxidative phosphorylation [61], and, under cellular stress, cannabinoids protect mitochondria [62], which together could be essential for the survival of noradrenergic neurons. Cellular stress resistance is largely dependent on cell metabolism and also on mitochondrial function. ...
Our laboratory and others have previously shown that cannabinoid receptor type-1 (CB1r) activity is neuroprotective and a modulator of brain ageing; a genetic disruption of CB1r signaling accelerates brain ageing, whereas the pharmacological stimulation of CB1r activity had the opposite effect. In this study, we have investigated if the lack of CB1r affects noradrenergic neurons in the locus coeruleus (LC), which are vulnerable to age-related changes; their numbers are reduced in patients with neurodegenerative diseases and probably also in healthy aged individuals. Thus, we compared LC neuronal numbers between cannabinoid 1 receptor knockout (Cnr1-/-) mice and their wild-type littermates. Our results reveal that old Cnr1-/- mice have less noradrenergic neurons compared to their age-matched wild-type controls. This result was also confirmed by the analysis of the density of noradrenergic terminals which proved that Cnr1-/- mice had less compared to the wild-type controls. Additionally, we assessed pro-inflammatory glial activity in the LC. Although the density of microglia in Cnr1-/- mice was enhanced, they did not show enhanced inflammatory profile. We hypothesize that CB1r activity is necessary for the protection of noradrenergic neurons, but its anti-inflammatory effect probably only plays a minor role in it.
... CB1 is also localized on skeletal muscle mitochondria where it directly controls cellular respiration and ATP production, 25 and its activation is known to impair mitochondrial function and biogenesis. [25][26][27][28][29] This process is in part mediated through the production of ceramides, 29 lipid derivatives associated with the development of lipotoxicity and insulin resistance commonly seen in sarcopenia. 5 Lastly, CB1 overactivation is also associated with skeletal muscle development and function. ...
... Through this interaction, AEA mediates cellular death in rat and human neurons [46,47], lymphoma cells (47) and cytotrophoblasts (48) . Ligands of cannabinoid receptors generally induce apoptosis inhibiting mitochondrial activity because they increase mitochondrial hydrogen peroxide production and decrease the consumption of oxygen and the mitochondrial membrane potential (49) . Another important effect of cannabinoid receptor activation is the inhibition of cancer cells invasion. ...
Objectives: The endocannabinoid system (ECS) is made up of an array of endogenous bioactive lipids, their receptors and enzymes for their synthesis and degradation. The main endogenous ligands are unsaturated fatty acid derivatives such as anandamide (AEA) and 2-arachidonoylglycerol (2-AG), but many others are still under study. Endocannabinoids are involved in both physiological and pathological conditions and could play an important role in the regulation of processes which lead to cancer.
Methods: With focus on gynaecological cancers, main papers and review articles, up to September 2018, on the role of the ECS, were acquired by PubMed searches using the search terms: ‘cannabinoid’, ‘endocannabinoid’, ‘gynaecology’, ‘cancer’, and ‘malignancy’.
Results: The present review showed the involvement of the endocannabinoid system in numerous physiological and pathological conditions of the female genital tract up to the development of gynaecological malignancy as cervical, endometrial and ovarian cancer.
Conclusion: The endocannabinoid system has an important role in antitumor actions involving different signalling receptor and receptor-independent pathways. It represents an exciting challenge to researchers for its potential use in diagnosis and treatment of all gynaecological malignancies.
... Siegmund reported that the CB 2-AG stimulation increases mitochondrial ROS productions and mitochondrial depolarization in hepatic stellate cells. Different other CB receptor agonists (anandamide, phytocannabinoid D-9-tetrahydrocannabinol, and synthetic CB HU 210) have also been reported to decrease mitochondrial membrane potential and increase mitochondrial ROS production (Athanasiou et al., 2007). Interestingly, in a recent report, ROS has been shown to be involved in MK maturation . ...
Endocannabinoids are well‐known regulators of neurotransmission by activating the cannabinoid (CB) receptors. Endocannabinoids are being used extensively for the treatment of various neurological disorders such as Alzheimer's and Parkinson's diseases. Although endocannabinoids are well studied in cell survival, proliferation, and differentiation in various neurological disorders and several cancers, the functional role in the regulation of blood cell development is less examined. In the present study, virodhamine, which is an agonist of CB receptor‐2, was used to examine its effect on megakaryocytic development from a megakaryoblastic cell. We observed that virodhamine increases cell adherence, cell size, and cytoplasmic protrusions. Interestingly, we have also observed large nucleus and increased expression of megakaryocytic marker (CD61), which are the typical hallmarks of megakaryocytic differentiation. Furthermore, the increased expression of CB2 receptor was noticed in virodhamine‐induced megakaryocytic cells. The effect of virodhamine on megakaryocytic differentiation could be mediated through CB2 receptor. Therefore, we have studied virodhamine induced molecular regulation of megakaryocytic differentiation; mitogen‐activated protein kinase (MAPK) activity, mitochondrial function, and reactive oxygen species (ROS) production were majorly affected. The altered mitochondrial functions and ROS production is the crucial event associated with megakaryocytic differentiation and maturation. In the present study, we report that virodhamine induces megakaryocytic differentiation by triggering MAPK signaling and ROS production either through MAPK effects on ROS‐generating enzymes or by the target vanilloid receptor 1‐mediated regulation of mitochondrial function. Virodhamine, an endocannabinoid, induces megakaryocyte differentiation by regulating mitogen‐activated protein kinase activity and function of mitochondria.
... While it has been demonstrated that THC impairs mitochondrial function in neuronal systems (Athanasiou et al., 2007;Wolff et al., 2015), the consequences of THC on trophoblast mitochondrial dynamics, respiration, and associated stress responses have not been fully explored. Given that mitochondrial dysfunction, oxidative stress, and poor trophoblast outcomes (Walker et al., 2020), and THC exposure during pregnancy leading to poor placentation and restricted intrauterine growth (Natale et al., 2020) has been demonstrated in rats, we hypothesize that THC may directly increase oxidative stress and reduce ATP generation, and alter trophoblast gene expression resulting in attenuated syncytialization. ...
The psychoactive component in cannabis, delta-9-tetrahydrocannabinol, can restrict fetal growth and development. Delta-9-tetrahydrocannabinol has been shown to negatively impact cellular proliferation and target organelles like the mitochondria resulting in reduced cellular respiration. In the placenta, mitochondrial dysfunction leading to oxidative stress prevents proper placental development and function. A key element of placental development is the proliferation and fusion of cytotrophoblasts to form the syncytium that comprises the materno-fetal interface. The impact of delta-9-tetrahydrocannabinol on this process is not well understood. To elucidate the nature of the mitochondrial dysfunction and its consequences on trophoblast fusion, we treated undifferentiated and differentiated BeWo human trophoblast cells, with 20 µM delta-9-tetrahydrocannabinol for 48 hr. At this concentration, delta-9-tetrahydrocannabinol on BeWo cells reduced the expression of markers involved in syncytialization and mitochondrial dynamics, but had no effect on cell viability. Delta-9-tetrahydrocannabinol significantly attenuated the process of syncytialization and induced oxidative stress responses in BeWo cells. Importantly, delta-9-tetrahydrocannabinol also caused a reduction in the secretion of human chorionic gonadotropin and the production of human placental lactogen and insulin growth factor 2, three hormones known to be important in facilitating fetal growth. Furthermore, we also demonstrate that delta-9-tetrahydrocannabinol attenuated mitochondrial respiration, depleted adenosine triphosphate, and reduced mitochondrial membrane potential. These changes were also associated with an increase in cellular reactive oxygen species, and the expression of stress responsive chaperones, HSP60 and HSP70. These findings have important implications for understanding the role of delta-9-tetrahydrocannabinol-induced mitochondrial injury and the role this might play in compromising human pregnancies.
... For instance, Beńard et al. (2012) used anti-CB1R antibodies, disclosed the protein localization of CB1R nearly 30 percent of neuronal mitochondria, which when triggered by exogenous/endogenous cannabinoids lowers the respiratory chain complex-I activity and oxygen consumption, probably via cyclic adenosine monophosphate (cAMP) and protein kinase-A (PKA) signaling. These results are supported by Athanasiou et al. (2007) findings, which reveal that all of the partial CB1R agonists including AEA, D-9-THC, and HU-210 markedly reduced oxygen consumption ( Figure 3) and mitochondrial membrane potential. However, care must be taken to interpret these findings due to using commercial anti-CB1R antibodies (Morozov et al., 2013). ...
Alzheimer’s disease (AD) is an irreversible chronic neurodegenerative disorder that occurs when neurons in the brain degenerate and die. Pain frequently arises in older patients with neurodegenerative diseases including AD. However, the presence of pain in older people is usually overlooked with cognitive dysfunctions. Most of the times dementia patients experience moderate to severe pain but the development of severe cognitive dysfunctions tremendously affects their capability to express the presence of pain. Currently, there are no effective treatments against AD that emphasize the necessity for increasing research to develop novel drugs for treating or preventing the disease process. Furthermore, the prospective therapeutic use of cannabinoids in AD has been studied for the past few years. In this regard, targeting the endocannabinoid system has considered as a probable therapeutic strategy to control several associated pathological pathways, such as mitochondrial dysfunction, excitotoxicity, oxidative stress, and neuroinflammation for the management of AD. In this review, we focus on recent studies about the role of cannabinoids for the treatment of pain and related neuropathological changes in AD.
... CB 1 R agonists have also been reported to decrease temperature (25,49), which could potentially be harmful and exacerbate immune dysfunction in sepsis (50,51). Conversely, D9-THC has been reported to decrease oxygen consumption (52,53), suggesting that it might be protective in conditions in which there is low oxygen delivery such as septic shock and ischemiareperfusion injury. We also observed that D9-THC reduces LPSinduced tachycardia in the first hours following injection. ...
Cannabis sativa and its principal components, Δ9-tetrahydrocannabinol (Δ9-THC) and cannabidiol, are increasingly being used to treat a variety of medical problems, including inflammatory conditions. Although studies suggest that the endocannabinoid system has immunomodulatory properties, there remains a paucity of information on the effects of cannabinoids on immunity and on outcomes of infection and injury. We investigated the effects and mechanism(s) of action of cannabinoid receptor agonists, including Δ9-THC, on inflammation and organ injury in endotoxemic mice. Administration of Δ9-THC caused a dramatic early upregulation of plasma IL-10 levels, reduced plasma IL-6 and CCL-2 levels, led to better clinical status, and attenuated organ injury in endotoxemic mice. The anti-inflammatory effects of Δ9-THC in endotoxemic mice were reversed by a cannabinoid receptor type 1 (CB1R) inverse agonist (SR141716), and by clodronate-induced myeloid-cell depletion, but not by genetic invalidation or blockade of other putative Δ9-THC receptors, including cannabinoid receptor type 2, TRPV1, GPR18, GPR55, and GPR119. Although Δ9-THC administration reduced the activation of several spleen immune cell subsets, the anti-inflammatory effects of Δ9-THC were preserved in splenectomized endotoxemic mice. Finally, using IL-10-GFP reporter mice, we showed that blood monocytic myeloid-derived suppressive cells mediate the Δ9-THC-induced early rise in circulating IL-10. These results indicate that Δ9-THC potently induces IL-10, while reducing proinflammatory cytokines, chemokines, and related organ injury in endotoxemic mice via the activation of CB1R. These data have implications for acute and chronic conditions that are driven by dysregulated inflammation, such as sepsis, and raise the possibility that CB1R-signaling may constitute a novel target for inflammatory disorders.
Aims:
This study established the in vitro anti-lymphoma pharmacodynamic actions of the endocannabinoids (anandamide-AEA and 2-arachidonoylglycerol-2AG) on canine non-Hodgkin lymphoma (NHL) and human NHL cells.
Main methods:
The expression of cannabinoid (CB1 and CB2) receptors in various canine NHL cells {1771, CLBL-1, CLL-1, peripheral blood mononuclear cells (PBMCs)} was studied using Quantitative real-time PCR (RT-qPCR). Anti-lymphoma cell viability assay was performed to assess the effect of endocannabinoids on various canine and human NHL cells (1771, CLBL-1, CLL-1, Ramos cells). The spectrophotometric and fluorometric procedures evaluated oxidative stress, inflammation, apoptosis, and mitochondrial function markers. SAS® and Prism-V La Jolla, CA, USA, were used for statistical analysis.
Key findings:
The current study validated the presence of CB1 and CB2 receptors in the canine NHL cells. There was a significantly higher expression of CB1 and CB2 receptors in B-cell lymphoma (BCL) cells (1771, CLBL-1, Ramos) compared to canine T-cell lymphoma (TCL) cells (CL-1). AEA and 2AG dose and time-dependently exhibited significant but differential anti-lymphoma effects on canine and human NHL cells. Anti-lymphoma pharmacodynamic actions of the endocannabinoids in the canine 1771 NHL cells revealed a significant alteration in the markers of oxidative stress, inflammation, and a decrease in mitochondrial function without altering the apoptotic markers.
Significance:
Establishing the anti-lymphoma pharmacodynamic actions of endocannabinoids may provide new therapeutic interventions and expedite cannabinoid research.
The medicinal properties of cannabis and cannabinoid-derivative are entirely investigated and known. In addition, the identification of psychotropic plant cannabinoids has led to more studies regarding the cannabinoid system and its therapeutic features in the treatment and management of clinical symptoms of neuroinflammatory disorders, such as multiple sclerosis (MS), Parkinsons disease (PD), and Alzheimers disease (AD). In fact, cannabinoid agonists are able to control and regulate inflammatory responses. In contrast to the cannabinoid receptor type 1 (CB1) and its unwanted adverse effects, the cannabinoid receptor type 2 (CB2) and its ligands hold promise for new and effective therapeutic approaches. So far, some successes have been achieved in this field. This review will discuss an outline of the endocannabinoid system's involvement in neuroinflammatory disorders. Moreover, the pharmacological efficacy of different natural and synthetic preparations of phytocannabinoids acting on cannabinoid receptors, particularly in MS, PD, and AD, will be updated. Also, the reasons for targeting CB2 for neurodegeneration will be explained.
Mitochondria play a critical role in the regulation of several biological processes (e.g., programmed cell death, inflammation, neurotransmission, cell differentiation). In recent years, accumulating findings have evidenced that cannabinoids, a group of endogenous and exogenous (synthetic and plant-derived) psychoactive compounds that bind to cannabinoid receptors, may modulate mitochondrial function and dynamics. As such, mitochondria have gained increasing interest as central mediators in cannabinoids’ pharmacological and toxicological signatures.
Here, we review the mechanisms underlying the cannabinoids’ modulation of mitochondrial activity and dynamics, as well as the potential implications of such mitochondrial processes’ disruption on cell homeostasis and disease. Interestingly, cannabinoids may target different mitochondrial processes (e.g., regulation of intracellular calcium levels, bioenergetic metabolism, apoptosis, and mitochondrial dynamics, including mitochondrial fission and fusion, transport, mitophagy, and biogenesis), by modulating multiple and complex signaling pathways. Of note, the outcome may depend on the experimental models used, as well as the chemical structure, concentration, and exposure settings to the cannabinoid, originating equivocal data. Notably, this interaction seems to represent not only an important feature of cannabinoids’ toxicological signatures, with potential implications for the onset of distinct pathological conditions (e.g., cancer, neurodegenerative diseases, metabolic syndromes), but also an opportunity to develop novel therapeutic strategies for such pathologies, which is also discussed in this review.
COVID-19 is caused by SARS-CoV-2 and leads to acute lung injury (ALI), acute respiratory distress syndrome (ARDS), and extrapulmonary manifestations in severely affected cases. However, most of the affected cases are mild or asymptomatic. Cannabinoids (CBs) such as tetrahydrocannabinol (THC) and cannabidiol (CBD), which act on G-protein-coupled receptors called CB1 and CB2, have anti-inflammatory effects. Many published studies show that CBs are effective in various inflammatory disorders, viral infections, and attenuation of ALI and ARDS. Therefore, the aim of the present narrative review was to summarize the possible immunological role of CBs in COVID-19. The effects of CBs are controversial, although they have beneficial effects via CB2 receptors and adverse effects via CB1 receptors against ALI, ARDS, and hyperinflammation, which are hallmarks of COVID-19. The present narrative review has shown that CBs effectively manage ALI and ARDS by suppressing pro-inflammatory cytokines, which are common in COVID-19. Therefore, CBs may be used to manage COVID-19 because of their potent anti-inflammatory effects with suppression of pro-inflammatory cytokines and inhibition of inflammatory signaling pathways.
Diabetic complications, chiefly seen in long-term situations, are persistently deleterious to a large extent, requiring multi-factorial risk reduction strategies beyond glycemic control. Diabetic cardiomyopathy is one of the most common deleterious diabetic complications, being the leading cause of mortality among diabetic patients. The mechanisms of diabetic cardiomyopathy are multi-factorial, involving increased oxidative stress, accumulation of advanced glycation end products (AGEs), activation of various pro-inflammatory and cell death signaling pathways, and changes in the composition of extracellular matrix with enhanced cardiac fibrosis. The novel lipid signaling system, the endocannabinoid system, has been implicated in the pathogenesis of diabetes and its complications through its two main receptors: Cannabinoid receptor type 1 and cannabinoid receptor type 2, alongside other components. However, the role of the endocannabinoid system in diabetic cardiomyopathy has not been fully investigated. This review aims to elucidate the possible mechanisms through which cannabinoids and the endocannabinoid system could interact with the pathogenesis and the development of diabetic cardiomyopathy. These mechanisms include oxidative/ nitrative stress, inflammation, accumulation of AGEs, cardiac remodeling, and autophagy. A better understanding of the role of cannabinoids and the endocannabinoid system in diabetic cardiomyopathy may provide novel strategies to manipulate such a serious diabetic complication.
Exploring molecular mechanisms of aging and determinants of lifespan will help reduce age-related morbidity, thus facilitating healthy brain aging. Recently, it has been demonstrated that nutritional polyphenols, the main constituents of the Mediterranean diet, maintain redox balance and neuroprotection through the activation of hormetic vitagene pathway. Mitochondria play pivotal roles in the mechanisms of cellular aging and lifespan extension, although further studies are required concerning optimal bioenergetic mechanisms promoting aerobic energy production and the underlying detrimental effects of reactive oxygen species (ROS) by-production with the interplayed nutrition and caloric intake modulatory effects. Consistently, ROS acting as sensors of intracellular nutrients and energy state regulate functional mitochondrial state. Interestingly, increasing evidence reports a functional crosstalk between ROS production by mitochondria and longevity pathways modulating lifespan across species thus ensuring healthy aging. Nrf2-dependent pathways of cellular stress response with their target antioxidant vitagenes are emerging as powerful systems capable to preserve redox homeostasis under environmental and metabolic stresses. Vitagenes encode redox longevity genes induced by oxidative damage including heat shock family (Hsp) Hsp32, Hsp70, glutathione, thioredoxin and sirtuin protein systems. During aging process, a gradual decline of the heat shock response occurs and this may prevent repair of protein damage. Therefore, there is a growing interest by scientific community in developing of novel preventive andpharmacological agents capable of inducing stress responses at the minimum dose within the broad frame of hormesis as therapeutic strategy in patients suffering from chronic degenerative diseases. The specialattention of this paper is focusedonpotential neuroprotective mechanisms of nutrition, in particular dietary polyphenols involved in the activation of vitagenes resulting in improved intracellular antioxidant defense systems against ROS damage leading to degeneration and death with considerable impact on brain health and longevity processes.
Cannabis psychoactive and medicinal properties have been known for thousands of years, but it is only recently that we started to understand the cellular mechanisms triggered by phytocannabinoids, such as Δ⁹-tetrahydrocannabinol and cannabidiol. The discovery of the endocannabinoid system, which includes cannabinoid receptors and cannabinoids produced endogenously in animal cells, allowed a better understanding of the impact of these phytocannabinoids on human health. Early findings suggested that these molecules disrupt mitochondrial functions via direct binding to the organelle membranes and subsequent alterations of their structure and functions. The multiple tools generated after the discovery of the cannabinoid receptors in the early 1990s allowed to show that specific cannabinoids can also modulate mitochondrial activity via activation of cannabinoid receptors localized at the plasma membrane or directly within mitochondrial membranes. Considering that mitochondria are involved in the onset and progression of various diseases, it appears crucial to better understand the mechanisms linking cannabinoids and mitochondria. The aim of this chapter is to provide an overview of the multiple mechanisms triggered by cannabinoids that affects mitochondria.
Toll-like receptors (TLRs) are sensors of pathogen-associated molecules that trigger inflammatory signalling in innate immune cells including macrophages. All TLRs, with the exception of TLR3, promote intracellular signalling via recruitment of the myeloid differentiation factor 88 (MyD88) adaptor, while TLR3 signals via Toll-Interleukin-1 Receptor (TIR)-domain-containing adaptor-inducing interferon (IFN)-β (TRIF) adaptor to induce MyD88-independent signalling. Furthermore, TLR4 can activate both MyD88-dependent and –independent signalling (via TRIF). The study aim was to decipher the impact of the highly purified plant-derived (phyto) cannabinoids Δ⁹-tetrahydrocannabinol (THC) and cannabidiol (CBD), when delivered in isolation and in combination (1:1), on MyD88-dependent and -independent signalling in macrophages. We employed the use of the viral dsRNA mimetic poly(I:C) and endotoxin lipopolysaccharide (LPS), to induce viral TLR3 and bacterial TLR4 signalling in human Tamm-Horsfall protein-1 (THP-1)-derived macrophages, respectively. TLR3/TLR4 stimulation promoted the activation of interferon (IFN) regulatory factor 3 (IRF3) and TLR4 promoted the activation of nuclear factor (NF)-κB signalling, with downstream production of the type I IFN-β, the chemokines CXCL10 and CXCL8, and cytokine TNF-α. THC and CBD (both at 10 μM) attenuated TLR3/4-induced IRF3 activation and induction of CXCL10/IFN-β, while both phytocannabinoids failed to impact TLR4-induced IκB-α degradation and TNF-α/CXCL8 expression. The role of CB1, CB2 and PPARγ receptors in mediating the effect of THC and CBD on MyD88-independent signalling was investigated. TLRs are attractive therapeutic targets given their role in inflammation and initiation of adaptive immunity, and data herein indicate that both CBD and THC preferentially modulate TLR3 and TLR4 signalling via MyD88-independent mechanisms in macrophages. This offers mechanistic insight into the role of phytocannabinoids in modulating cellular inflammation.
Aim:
The aim of this review article is to summarize current knowledge about the role of cannabinoids and cannabinoid receptors in tumor disease modulation and to evaluate comprehensively the use of cannabinoids in cancer patients.
Method:
According to the PRISMA protocol, we have included data from a total of 105 articles.
Results:
Cannabinoids affect cancer progression by three mechanisms. The most important mechanism is the stimulation of autophagy and affecting the signaling pathways leading to apoptosis. The most important mechanism of this process is the accumulation of ceramide. Cannabinoids also stimulate apoptosis by mechanisms independent of autophagy. Other mechanisms by which cannabinoids affect tumor growth are inhibition of tumor angiogenesis, invasiveness, metastasis, and the modulation of the anti-tumor immune response.
Conclusion:
In addition to the symptomatic therapy of cancer patients, the antitumor effects of cannabinoids (whether in monotherapy or in combination with other cancer therapies) have promising potential in the treatment of cancer patients. More clinical trials are needed to demonstrate the antitumor effect of cannabinoids (Tab. 1, Fig. 1, Ref. 167).
Delta9-Tetrahydrocannabinol (delta9-THC), the active ingredient of marihuana was found to be a highly effective inhibitor in vitro of the NADH-oxidase activity of rat brain and heart mitochondria. The degree of inhibition of the enzyme system obtained from rat brain tissue varied with the region from which it was derived as follows, in the presence of 10(-5) M delta9-THC: hypothalamus plus thalamus plus midbrain, 73 +/- 4%; cerebellum, 66 +/- 4%; medulla oblongata plus pons, 63 +/- 6%; cerebral cortex, 50 +/- 8%. The same concentration inhibited rat heart NADH-oxidase activity 69 +/- 9%. Inhibition of NADH-oxidase activity by a corresponding concentration of deoxycorticosterone was significantly less in all tissue preparations tested, ranging from 11% to 26%. The inhibition of delta9-THC appeared to be competitive and near the amytal-sensitive site of the electron transport system. Suggestive evidence was also obtained for a second site of action, above the cytochrome c site.
The role of endocannabinoid signaling in the response of the brain to injury is tantalizing but not clear. In this study, transient middle cerebral artery occlusion (MCAo) was used to produce ischemia/reperfusion injury. Brain content of N-arachidonoylethanolamine (AEA) and 2-arachidonoylglycerol were determined during MCAo. Whole brain AEA content was significantly increased after 30, 60 and 120 min MCAo compared with sham-operated brain. The increase in AEA was localized to the ischemic hemisphere after 30 min MCAo, but at 60 and 120 min, was also increased in the contralateral hemisphere. 2-Arachidonoylglycerol content was unaffected by MCAo. In a second set of studies, injury was assessed 24 h after 2 h MCAo. Rats administered a single dose (3 mg/kg) of the cannabinoid receptor type 1 (CB1) receptor antagonist SR141716 prior to MCAo exhibited a 50% reduction in infarct volume and a 40% improvement in neurological function compared with vehicle control. A second CB1 receptor antagonist, LY320135 (6 mg/kg), also significantly improved neurological function. The CB1 receptor agonist, WIN 55212-2 (0.1-1 mg/kg) did not affect either infarct volume or neurological score.
Mitochondria have been suggested to be a potential intracellular target for cancer chemotherapy. In this report, we demonstrate the ability of the tricyclic antidepressant chlorimipramine to kill human glioma cells in vitro by a molecular mechanism resulting in an increase in caspase 3 activity following inhibition of glioma oxygen consumption. Studies with isolated rat mitochondria showed that chlorimipramine specifically inhibited mitochondrial complex III activity, which causes decreased mitochondrial membrane potential as well as mitochondrial swelling and vacuolation. The use of chlorimipramine in human as an effective, non-toxic cancer therapeutic having a strong selectivity between cancer cells and normal cells on the basis of their mitochondrial function is discussed.
Habitual marijuana smoking is associated with inflammation and atypia of airway epithelium accompanied by symptoms of chronic bronchitis. We hypothesized that Delta(9)-tetrahydrocannabinol (THC), the primary psychoactive component of marijuana, might contribute to these findings by impairing cellular energetics and mitochondrial function. To test this hypothesis, we examined particulate smoke extracts from marijuana cigarettes, tobacco cigarettes, and placebo marijuana (0% THC) cigarettes for their effects on the mitochondrial function of A549 cells in vitro. Only extracts prepared from marijuana cigarettes altered mitochondrial staining by the potentiometric probe JC-1. With the use of a cross-flow, nose-only inhalation system, rats were then exposed for 20 min to whole marijuana smoke and examined for its effects on airway epithelial cells. Inhalation of marijuana smoke produced lung tissue concentrations of THC that were 8-10 times higher than those measured in blood (75 +/- 38 ng/g wet wt tissue vs. 9.2 +/- 2.0 ng/ml), suggesting high local exposure. Intratracheal infusion of JC-1 immediately following marijuana smoke exposure revealed a diffuse decrease in lung cell JC-1 red fluorescence compared with tissue from unexposed or placebo smoke-exposed rats. Exposure to marijuana smoke in vivo also decreased JC-1 red fluorescence (54% decrease, P < 0.01) and ATP levels (75% decrease, P < 0.01) in single-cell preparations of tracheal epithelial cells. These results suggest that inhalation of marijuana smoke has deleterious effects on airway epithelial cell energetics that may contribute to the adverse pulmonary consequences of marijuana smoking.
The impaired mitochondrial function hypothesis in schizophrenia is based on evidence of altered brain metabolism, morphology, biochemistry and gene expression. Mitochondria have their own genome, which is needed to synthesize some of the subunits of the respiratory chain enzymes. Mitochondrial DNA (mtDNA) is maternally inherited and we observed an excess of maternal transmission of schizophrenia in a set of parent-offspring affected pairs. We therefore hypothesized that mutations in the mtDNA may contribute to the complex genetic basis of schizophrenia. The entire mtDNA of six schizophrenic patients with an apparent maternal transmission of the disease was sequenced and compared to the reference sequence. We have identified 50 variants and among these six have not been previously reported. Three of them were missense variants: MTCO2 7750C>A, MTATP6 8857G>A and MTND4 12096T>A. These were maternally inherited because they were also present in the mtDNA of their respective schizophrenic mothers and none of them were found in 95 control individuals. The MTND4 12096T>A (Leu446His) is a heteroplasmic variant present in five of the six mother-offspring patient pairs that triggers a non-conservative substitution in the ND4 subunit of complex I. Sequence alignment of 110 ND4 peptides from all eukaryotic kingdoms shows that only hydrophobic amino acids are found in this position. Moreover, leucine was conserved or substituted by an isoleucine in all mammalian species. This indicates that the presence of histidine could affect complex I activity in patients with schizophrenia.
Mammalian tissues express at least two types of cannabinoid receptor, CB1 and CB2, both G protein coupled. CB1 receptors are expressed predominantly at nerve terminals where they mediate inhibition of transmitter release. CB2 receptors are found mainly on immune cells, one of their roles being to modulate cytokine release. Endogenous ligands for these receptors (endocannabinoids) also exist. These are all eicosanoids; prominent examples include arachidonoylethanolamide (anandamide) and 2-arachidonoyl glycerol. These discoveries have led to the development of CB1- and CB2-selective agonists and antagonists and of bioassays for characterizing such ligands. Cannabinoid receptor antagonists include the CB1-selective SR141716A, AM251, AM281 and LY320135, and the CB2-selective SR144528 and AM630. These all behave as inverse agonists, one indication that CB1 and CB2 receptors can exist in a constitutively active state. Neutral cannabinoid receptor antagonists that seem to lack inverse agonist properties have recently also been developed. As well as acting on CB1 and CB2 receptors, there is convincing evidence that anandamide can activate transient receptor potential vanilloid type 1 (TRPV1) receptors. Certain cannabinoids also appear to have non-CB1, non-CB2, non-TRPV1 targets, for example CB2-like receptors that can mediate antinociception and "abnormal-cannabidiol" receptors that mediate vasorelaxation and promote microglial cell migration. There is evidence too for TRPV1-like receptors on glutamatergic neurons, for alpha2-adrenoceptor-like (imidazoline) receptors at sympathetic nerve terminals, for novel G protein-coupled receptors for R-(+)-WIN55212 and anandamide in the brain and spinal cord, for novel receptors for delta9-tetrahydrocannabinol and cannabinol on perivascular sensory nerves and for novel anandamide receptors in the gastro-intestinal tract. The presence of allosteric sites for cannabinoids on various ion channels and non-cannabinoid receptors has also been proposed. In addition, more information is beginning to emerge about the pharmacological actions of the non-psychoactive plant cannabinoid, cannabidiol. These recent advances in cannabinoid pharmacology are all discussed in this review.
It is now well established that the psychoactive effects of Cannabis sativa are primarily mediated through neuronal CB1 receptors, while its therapeutic immune properties are primarily mediated through CB2 receptors. Two endocannabinoids, arachidonoylethanolamide and 2-arachidonoylglycerol, have been identified, their action on CB1 and CB2 thoroughly characterized, and their production and inactivation elucidated. However, many significant exceptions to these rules exist. Here we review the evidence suggesting that cannabinoids can modulate synaptic transmission, the cardiovascular system, and the immune system through receptors distinct from CB1 and CB2, and that an additional "independent" endocannabinoid signaling system that involves palmitoylethanolamide may exist.
Early and late effects of alloxan diabetes and insulin treatment on mitochondrial membrane structure and function were evaluated by studying the kinetic properties of mitochondrial membrane marker enzyme FoF1-ATPase and its modulation by membrane lipid/phospholipid composition and membrane fluidity. Under all experimental conditions the enzyme displayed three kinetically distinguishable components. In 1 wk-old diabetic animals the enzyme activity was unchanged; however, K(m) and V(max) of component I increased and K(m) of component II decreased. Insulin treatment resulted in lowering of K(m) and V(max) of components II and Ill. One-mon diabetic state resulted in decreased enzyme activity, whereas insulin treatment caused hyperstimulation. K(m) of components I and II decreased together with decreased V(max) of all the components. Insulin treatment restored the K(m) and V(max) values. In late-stage diabetes the catalytic efficiency of components I and II increased; insulin treatment had drastic adverse effect. Binding pattern of ATP was unchanged under all experimental conditions. Diabetic state resulted in progressive decrease in energy of activation in the low temperature range (E(L)). Insulin treatment lowered the energy of activation in the high temperature range (E(H)) without correcting the E(L) values. The phase transition temperatures increased in diabetic state and were not corrected by insulin treatment. Long-term diabetes lowered the total phospholipid content and elevated the cholesterol content; insulin treatment had partial restorative effect. The membrane fluidity decreased in general in diabetic condition and was not corrected by insulin treatment at late stage. Regression analysis studies suggest that specific phospholipid classes and/or their ratios may play a role in modulation of the enzyme activity.
Cannabinoids have been shown to possess anti-inflammatory and neuroprotective properties, which were proposed to occur mainly via activation of the G-protein-coupled receptor CB(1) (cannabinoid receptor 1). Recently, certain cannabinoids have been reported to be ligands for members of the nuclear receptor transcription factor superfamily known as PPARs (peroxisome-proliferator-activated receptors). This review summarizes the evidence for cannabinoid activation of PPARs and identifies a new intracellular target for cannabinoids as therapeutic agents for neuroprotective treatment.
Mitochondria play a pivotal role in the regulation of apoptotic cell death as well as in several cellular metabolic processes, including energy supply.1, 2 The latter is achieved by oxidative phosphorylation of ADP to ATP using the electrochemical proton gradient generated by the stepwise transport of electrons from oxidizable substrates to molecular oxygen mediated by the mitochondrial respiratory chain. One component of the respiratory chain is cytochrome c, which transfers electrons from Complex III to Complex IV. In apoptosis signaling, however, this vital function of cytochrome c is gradually lost.3 Once the outer mitochondrial membrane (OMM) has been permeabilized by proapoptotic members of the Bcl-2 family of proteins, cytochrome c is released from the mitochondrial intermembrane space into the cytosol. Here, it triggers apoptosome formation and the activation of the caspase cascade, which leads to the cleavage of a host of cellular proteins and dismantling of the cell (Figure 1). Within the mitochondria, cytochrome c is bound to the outer surface of the mitochondrial inner membrane (IMM) by its association with cardiolipin, an anionic phospholipid present predominantly in the mitochondria. We and others have previously suggested that the interaction of cytochrome c with cardiolipin critically determines the amount of the hemoprotein that can be released during apoptosis signaling.4, 5 Furthermore, there is emerging evidence that proapoptotic Bcl-2 family proteins might require cardiolipin for permeabilization of the mitochondria during apoptosis.6 However, the precise role of cardiolipin in the release of cytochrome c from mitochondria during apoptosis is still unclear and is the subject of this commentary.
Mammalian tissues express at least two types of cannabinoid receptor, CB1 and CB2, both G protein coupled. CB1 receptors are expressed predominantly at nerve terminals where they mediate inhibition of transmitter release. CB2 receptors
The major psychoactive component of marijuana, Δ9-tetrahydrocannabinol (THC), strongly affected rat liver mitochondria in vitro. At concentrations of 15–60 nmoles/mg of mitochondrial protein, THC uncoupled state IV respiration and decreased respiratory control and ADP/O ratios. Energy-linked changes in fluorescence of 8-anilino-1-naphthalene sulfonate were prevented or reversed by THC. THC also produced large amplitude swelling of mitochondria and release of matrix enzymes. These effects were greatly potentiated by Mg2+. Likewise, flocculation of mixed phospholipid micelles by Mg2+ was potentiated greatly by low concentrations of THC. Studies with micelles prepared from purified phospholipids suggest that THC may specifically destabilize the cardiolipin in mixed micelles.
The present study was undertaken in an attempt to establish whether or not some of the many biochemical effects of cannabinoids could be explained by inhibition of energy generation. Of particular interest was the relationship of inhibition of DNA synthesis in L1210 murine leukemia cells by certain cannabinoids to possible effects on respiration. In studies using these cells, delta9-tetrahydrocannabinol (delta9-THC) and delta8-tetrahydrocannabinol (delta8-THC) were inactive when tested both in vitro and in vivo. Since these cannabinoids have previously been shown to be inhibitors of DNA synthesis in L1210 cells, there is no necessary relationship between inhibition of the two processes.
The effect of cannabinoid derivatives on thymidine-3H uptake in L1210 murine leukemia was determined. In experiments at 200 mg/kg 3 hrs after treatment, the order of activity was delta9-tetrahydrocannabinol less than cannabinol less than cannabidiol less than abnormal cannabidiol less than 11-hydroxy-delta9-tetrahydrocannabinol less than delta8-tetrahydrocannabinol. The inhibitory effect of delta8-tetrahydrocannabinol was 99%. When animals were dosed on consecutive days with delta9-tetrahydrocannabinol and killed on the third day, thymidine-3H incorporation was increased while delta8-tetrahydrocannabinol retained its inhibitory activity under the same conditions. Delta-9-tetrahydrocannabinol and delta8-tetrahydrocannabinol inhibited RNA and protein synthesis in a fashion analagous to the inhibition of DNA synthesis.
affects the configurational integrity of rat liver mitochondria to a degree dependent upon its concentration. When concentration is increased, the mitochondria pass through three different morphological stages, which are visualized by electron microscopy. These stages are correlated with changes in oxygen uptake, ATPase activity and turbidity. Starting with control mitochondria, designated as Stage A, addition of 15 μg per mg protein results in maximum stimulation of oxygen uptake (Stage B). In the presence of 50 μg per mg protein this stimulation vanishes, the inner mitochondrial structure is disrupted and maximum values of swelling and ATPase activity are attained (Stage C). At higher concentrations, up to more than 100 μg per mg protein, both stimulation of ATPase activity and swelling diminish. These effects are proposed to be associated with bonding of the hydrophobic to some protein-phospholipid receptors in the mitochondrial membrane.
Four persons developed marked parkinsonism after using an illicit drug intravenously. Analysis of the substance injected by two of these patients revealed primarily 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine (MPTP) with trace amounts of 1-methyl-4-phenyl-4-propionoxy-piperidine (MPPP). On the basis of the striking parkinsonian features observed in our patients, and additional pathological data from one previously reported case, it is proposed that this chemical selectively damages cells in the substantia nigra.
The postnatal development of the complexes of the electron transport chain in isolated rat brain mitochondria were investigated. Nonsynaptosomal brain mitochondria were isolated from rats aged 1-60 days, and the activities of mitochondrial complexes I, II-III, IV, V and citrate synthase were measured. There was a significant increase in the activity of complex I from postnatal day 1 to day 21, and in the activities of complex II-III, complex IV and citrate synthase from postnatal day 1 to day 60. In contrast, the activity of complex V increased significantly between postnatal day 1 and day 10 where it attained adult levels. These data are consistent with the increasing demand for mitochondrial ATP production as the brain develops and as aerobic glycolysis becomes the major pathway for energy production.
The effects of 1-methyl-4-phenylpyridinium (MPP+) on the oxygen consumption, ATP production, H2O2 production, and mitochondrial NADH-CoQ1 reductase (complex I) activity of isolated rat brain mitochondria were investigated. Using glutamate and malate as substrates, concentrations of 10-100 microM MPP+ had no effect on state 4 (-ADP) respiration but decreased state 3 (+ADP) respiration and ATP production. Incubating mitochondria with ADP for 30 min after loading with varying concentrations of MPP+ produced a concentration-dependent decrease in H2O2 production. Incubation of mitochondria with ADP for 60 min after loading with 100 microM MPP+ caused no loss of complex I activity after washing of MPP+ from the mitochondrial membranes. These data are consistent with MPP+ initially binding specifically to complex I and inhibiting both the flow of reducing equivalents and the production of H2O2 by the mitochondrial respiratory chain, without irreversibly damaging complex I. However, mitochondria incubated with H2O2 in the presence of Cu2+ ions showed decreased complex I activity. This study provides additional evidence that cellular damage initiated by MPP+ is due primarily to energy depletion caused by specific binding to complex I, any increased damage due to free radical production by mitochondria being a secondary effect.
The postnatal development of the complexes of the electron transport chain in mitochondria isolated from rat brain synaptosomes was investigated. Synaptosomal brain mitochondria were isolated from rats aged 10-60 days, and the activities of mitochondrial complex I, complex II-III, complex IV and complex V were measured. There was a significant increase in the activity of II-III from day 10 to day 15 and complex IV from day 10 to day 21, thereafter the activities of complexes I-III and IV did not change significantly. The activity of complex I did not change significantly during the period 10-60 days post partum. In synaptic mitochondria, complex V activity was higher than in non-synaptic mitochondria, whereas the activity of complex I was lower than in non-synaptic mitochondria. These data show that the complexes of the respiratory chain within synaptic mitochondria have activities different from those of non-synaptic mitochondria and may have major implications for the relative susceptibility of mitochondria in different brain cell types to neurotoxins such as MPP+, hypoxic/ischaemic damage and oxidative stress.
A colorimetric assay using the basic azo dye Janus green has been developed to assess cell numbers in anchorage-dependent cell cultures, with special regard to the enumeration of osteoblastic cells. Therefore, cells are fixed in ethanol and stained with a 0.2% solution of Janus green for 3 min, followed by a destaining step of 1 min in tap water. The addition of diluted hydrochloric acid easily and immediately leads to dye elution from stained cell layers into the acidic supernatant which consequently is transferred into 96-well plates and read on a microplate reader at 595 nm. Working under standardized conditions, Janus green uptake in several cell lines is shown to be linearly correlated with cell numbers over a broad range of cell densities, in MC3T3-E1 cells from about 3% up to more than 300% of confluency. Absolute sensitivity of the assay allows detection of less than 1000 cells/cm(2). In comparison to many other colorimetric assays, the Janus green technique is simple to perform, fast, precise, stable, cheap, and well suited for processing large quantities of samples. Moreover, it is applicable to any culture formate and size, from irregular formed carriers up to 96-multiwell plates.
It is becoming increasingly evident that the mitochondrial genome may play a key role in neurodegenerative diseases. Mitochondrial dysfunction is characteristic of several neurodegenerative disorders, and evidence for mitochondria being a site of damage in neurodegenerative disorders is partially based on decreases in respiratory chain complex activities in Parkinson's disease, Alzheimer's disease, and Huntington's disease. Such defects in respiratory complex activities, possibly associated with oxidant/antioxidant balance perturbation, are thought to underlie defects in energy metabolism and induce cellular degeneration. Efficient functioning of maintenance and repair process seems to be crucial for both survival and physical quality of life. This is accomplished by a complex network of the so-called longevity assurance processes, which are composed of genes termed vitagenes. A promising approach for the identification of critical gerontogenic processes is represented by the hormesis-like positive effect of stress. In the present review, we discuss the role of energy thresholds in brain mitochondria and their implications in neurodegeneration. We then review the evidence for the role of oxidative stress in modulating the effects of mitochondrial DNA mutations on brain age-related disorders and also discuss new approaches for investigating the mechanisms of lifetime survival and longevity.
The central nervous system is one of the main target organs in cyanide toxicity. In this study, primary cultures of chick embryonic neurons were used to characterize sodium cyanide (NaCN)-induced cell death and to investigate the mechanism of NaCN-mediated preconditioning. After treatment of the cells with 1mM NaCN for 1h followed by a NaCN-free incubation period of 23 h, we observed features of apoptosis such as a reduction in nuclear size, chromatin condensation and nuclear fragmentation as evaluated by nuclear staining with Hoechst 33258 and electron microscopy. In addition, NaCN-induced neurotoxicity was reduced by the protein synthesis inhibitor cycloheximide (CHX) suggesting an active type of cell death. Most of the neurons with condensed chromatin and a shrunken nuclei also showed membrane damage at a late stage. Mitochondrial membrane potential as well as the protein levels of Bcl-2 and Bcl-x(L) decreased 15-60 min and 1-3 h after the exposure to NaCN (1mM, 1h), respectively. Preconditioning caused by incubating chick neurons with 100 microM NaCN for 30 min followed by a NaCN-free interval of 24h significantly protected the neurons against subsequent NaCN (1mM, 1h)-induced damage. Preconditioning prevented NaCN-induced decrease in the mitochondrial membrane potential as well as in the protein levels of Bcl-2 and Bcl-x(L) suggesting that preconditioning-induced neuroprotection is mediated by preserving mitochondrial function.
This study presents a new animal model, the Large White Pig, which was tested for studying cannabinoids metabolism. The first step has focused on determination of plasma kinetics after injection of Delta(9)-tetrahydrocannabinol (THC) at different dosages. Seven pigs received THC by intravenous injections (50, 100 or 200 microg/kg). Plasma samples were collected during 48 h. Determination of cannabinoids concentrations were performed by gas chromatography/mass spectrometry. Results showed that plasma kinetics were comparable to those reported in humans. Terminal half-life of elimination was 10.6 h and a volume of distribution of 32 l/kg was calculated. In a second step, this model was used to determine the kinetic profile of cannabinoids distribution in tissues. Eight Large White male pigs received an injection of THC (200 microg/kg). Two pigs were sacrificed 30 min after injection, two others after 2, 6 and 24 h. Different tissues were sampled: liver, kidney, heart, lung, spleen, muscle, fat, bile, blood, vitreous humor and several brain areas. The fastest THC elimination was noted in liver tissue, where it was completely eliminated in 6 h. THC concentrations decreased in brain tissue slower than in blood. The slowest THC elimination was observed for fat tissue, where the molecule was still present at significant concentrations 24 h later. After 30 min, THC concentration in different brain areas was highest in the cerebellum and lowest in the medulla oblongata. THC elimination kinetics noted in kidney, heart, spleen, muscle and lung were comparable with those observed in blood. 11-Hydroxy-THC was only found at high levels in liver. THC-COOH was less than 5 ng/g in most tissues, except in bile, where it increased for 24 h following THC injection. This study confirms, even after a unique administration, the prolonged retention of THC in brain and particularly in fat, which could be at the origin of different phenomena observed for heavy users such as prolonged detection of THC-COOH in urine or cannabis-related flashbacks. Moreover, these results support the interest for this animal model, which could be used in further studies of distribution of cannabinoids in tissues.
Time-lapse photomicroscopy of human H460 lung cancer cells demonstrated of the transient receptor potential V1 (TRPV1) channel agonists, (E)-capsaicin and resiniferatoxin, and the TRPV1 antagonists, capsazepine, and SB366791, were able to bring about morphological changes characteristic of apoptosis and/or necrosis. Immunoblot analysis identified immunoreactivity for the transient receptor potential V1 (TRPV1) channel in rat brain samples, but not in rat heart mitochondria or in H460 cells. In isolated rat heart mitochondria, all four ligands caused concentration-dependent decreases in oxygen consumption and mitochondrial membrane potential. (E)-Capsaicin and capsazepine evoked concentration-dependent increases and decreases, respectively, in mitochondrial hydrogen peroxide production, whilst resiniferatoxin and SB366791 were without significant effect. These data support the hypothesis that (E)-capsaicin, resiniferatoxin, capsazepine, and SB366791 are all mitochondrial inhibitors, able to activate apoptosis and/or necrosis via non-receptor mediated mechanisms, and also support the use of TRPV1 ligands as anti-cancer agents.
- C I Ragan
- Nadh-Ubiquinone Oxidoreductase
C.I. Ragan, NADH-ubiquinone oxidoreductase, Biochim. Biophys.
Acta 456 (1976) 249–290.
Chronic Parkinsonism in humans due to a product of meperidine-analog synthesis
- J W Langston
- P Ballard
- J W Tetrud
- I Irwin
J.W. Langston, P. Ballard, J.W. Tetrud, I. Irwin, Chronic Parkinsonism in humans due to a product of meperidine-analog synthesis,
Science 219 (1983) 979-980.
Chronic Parkinsonism in humans due to a product of meperidine-analog synthesis
- Langston