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Anti-inflammatory effects of aqueous extract from Dichroa febrifuga root in rat liver
Abstract
To study the anti-inflammatory effects of aqueous extract from Dichroa febrifuga root (AEDF) for suppression in the process of lipopolysaccharide (LPS)-induced sepsis in the rat liver.
The inhibitory effect of AEDF on the alteration of inflammatory proteins was investigated by Western blot and immunohistochemical analysis.
Western blot analysis showed that the level of nuclear factor (NF)-kappaBp65 was markedly up-regulated and (I)-kappaBalpha was down-regulated by LPS (8 mg/kg) challenge. However, AEDF 100 mg/kg inhibited induction of NF-kappaBp65 and degradation of I-kappaBalpha in the liver of LPS-challenged rats. Immunohistochemical analysis showed that while the expression of the NF-kappaBp65, tumor necrosis factor (TNF)-alpha and inducible nitric oxide synthase (iNOS) tended to increase, that of I-kappaBalpha was decreased in the hepatocytes of rats challenged with LPS. A slight decline of NF-kappaBp65, TNF-alpha and iNOS, but an increase of I-kappaBalpha were observed in the hepatocytes of the rats pretreated with AEDF.
AEDF may act as a therapeutic agent for inflammatory disease through a regulation of inflammation-related proteins.
... Bahkan, jenis tersebut juga dimanfaatkan sebagai obat batuk di China dan Korea. Selain itu, D. febrifuga juga dimanfaatkan sebagai bahan pelengkap pengobatan demam yang disebabkan oleh infeksi (Choi et al., 2003). Ekstrak akar D. ferbrifuga juga dapat dikembangkan sebagai obat anti inflamasi (Kim et al., 2000). ...
Kebun Raya Cibodas (KRC) sebagai kawasan konservasi ex-situ memiliki berbagai koleksi tumbuhan, salah satunya tumbuhan berpotensi obat. Inventarisasi jenis tumbuhan koleksi yang berpotensi antimalaria telah dilakukan melalui studi literatur dan penelusuran data koleksi tanaman KRC. Sebanyak 107 jenis tumbuhan koleksi KRC dari 50 suku tumbuhan berpotensi memiliki aktivitas antimalaria, diantaranya Alstonia scholaris (L) R. Br., Artemisia annua L., Azadirachta indica A. Juss., Bidens pilosa L., Cinchona calisaya Wedd., Dichroa febrifuga Lour.dan Vernonia amygdalina Del. Bagian tumbuhan yang menghasilkan zat antimalaria berasal dari ekstrak akar, batang, kulit batang dan daun. Jenis-jenis tumbuhan berpotensi antimalaria akan dijelaskan pada tulisan ini.
Kata kunci: Antimalaria, Kebun Raya Cibodas, obat alami, tumbuhan potensi obat.
Cibodas Botanic Gardens (CBG) as an ex-situ conservation area has a wide collection of plants, one of them is potential as medicinal plants. Inventory of plant species that potentially antimalarial collection has been conducted through literature study and CBG plants collection database searches. A total of 107 species of plants collection CBG from 50 families potentially have antimalarial activity, among others Alstonia scholaris (L) R. Br., Artemisia annua L., Azadirachta indica A. Juss., Bidens pilosa L., Cinchona calisaya Wedd., Dichroa febrifuga Lour., and Vernonia amygdalina Del. Parts of the plant which produces the antimalarial derived from the extract of the roots, stems, barks and leaves. The plants species potentially antimalarial will be explained in this paper.
Keywords: Antimalarial, Cibodas Botanic Gardens, herbal medicine, medicinal plant potential
... Halofuginone and extracts of Dichroa febrifuga have been shown to exhibit anti-inflammatory effects [21,29]. Although a gradual decrease (P>0.05) in the plasma level of PGM, an indicator of inflammation, was detected during halofuginone treatment, no statistically significant change in the plasma level of PGM was observed during the treatment period in the current study (Graphic 3). ...
Aims: The primary aim of this study was to evaluate the effect of halofuginone on serum ofthiobarbituric acid reactive substances (TBARS) and plasma testosterone, and 13,14-dihydro-15- keto-prostaglandinF2α (PGM) levels in male yearling sheep. It was also evaluated the effects of halofuginone on routine blood biochemistry and hemogram values.
Methodology: Ten male yearling sheep were treated at a dose of 0.1 mg/kg of halofuginone (PO, SID) for 15 days. Blood samples were collected before treatment (day 0, control) and on treatment days 5, 10, and 15. Hemogram parameters and the levels of TBARS, PGM, troponin I, creatinine kinase-MB, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transferase, total protein, blood urea nitrogen, creatinine, and calcium were determined.
Results: No statistically significant differences (P>0.05) in the levels of TBARS, testosterone or PGM were observed. The erythrocyte count, hematocrit, hemoglobin, total protein and calcium levels were lower (P<0.05) following the halofuginone treatment, whereas levels of troponin I, aspartate aminotransferase and alanine aminotransferase were higher (P<0.05), compared with the control values.
Conclusion: Halofuginone might cause anemia and organ damage, whereas it has no significant effect on indicators of fertility, oxidative status, or inflammation at the dosage and duration of treatment in male yearling sheep.
Nowadays, the surge of consumption of herbal supplements is encouraged by several factors, including the common belief that all herbal products are relatively safe and effective. The present investigation explores the effects of methanolic extract of Quercus infectoria bark upon rat blood lipid profile, glycemia, inflammation, gastric ulcer and bacterial growth. After one month of chronic extract (0.5% w/v) intake via drinking water, there was a significant increase in serum HDL-cholesterol level. This was accompanied with an increase in both serum glucose and insulin levels. No significant changes were observed in other lipid parameters studied. Liver enzyme activities as well as urea and creatinine levels were not negatively affected. Extract at 250, 500 and 1000 mg/kg body weight exhibited substantial anti-inflammatory effects in cases of acute and chronic inflammation induced by carrageenan and formalin respectively. Pre-treatment of fasted rats with the extract (100 and 500 mg/kg body weight) also demonstrated significant protection against ethanol-induced gastric ulcer. Anti-bacterial activity against Proteus mirabilis, Citrobacter braaki, and Staphylococcus aureus methicillin resistant and sensitive was also noticed. In conclusion, these findings suggest that the methanolic extract of Q. infectoria bark provides an inexpensive and powerful source of herbal supplement used to treat various conditions.
To investigate the effects of recombinant human growth hormone (rhGH) on rat acute lung injury (ALI).
ALI was mimicked by intraperitoneal (i. p.) injection of E. coli. Female Sprague-Dawley rats were randomized into three groups: control group, injected with physiologic saline (i. p.); ALI group, received a bolus injection of E. coli (1 x 10(10) cfu/l, 15 ml/kg, i. p.) followed by intramuscular physiologic saline injection; and ALI + GH group, received a bolus administration of E. coli, and then treated with intramuscular rhGH injection (2.25 U/kg/d). ALI group and ALI + GH group were subdivided into day 1 and day 3 subgroups, respectively. Left lungs were lavaged and bronchial alveolar lavage fluid (BALF) was harvested. Lung injury score, lung wet-to-dry weight (W/D) ratio, percentage of neutrophils (PMNs) in BALF, lung permeability index (LPI), intercellular adhesion molecule-1(ICAM-1) expression and activation of nuclear factor-kappa B (NF-kappaB) in the lungs were determined.
(1) On day 1 and day 3, the lung injury score, lung W/D ratio, LPI and protein content in BALF were significantly higher in the ALI group than in the control and ALI + GH groups. rhGH attenuated lung injuries significantly. (2) Compared with the control group, the percentage of PMNs in BALF was elevated significantly in the ALI and ALI + GH groups, especially in ALI group. (3) Nuclear positive rate of NF-kappaB and ICAM-1 expression at the levels of protein and mRNA in the lung in the ALI group on day 1 and day 3 were higher than in the control group. rhGH diminished activation of NF-kappaB and expression of ICAM-1 in the lung markedly.
Treatment with rhGH can significantly attenuate lung injury in the endotoxemic rats, which may be attributed to the reduction of the expression of ICAM-1, the influence on the adhesion and activation of PMNs, the inhibition of the activation of NF-kappaB and the regulation of the transcription of certain proinflammatory cytokines.
The transcription factor NF-kappa B is sequestered in the cytoplasm by the inhibitor protein I kappa B alpha. Extracellular inducers of NF-kappa B activate signal transduction pathways that result in the phosphorylation and subsequent degradation of I kappa B alpha. At present, the link between phosphorylation of I kappa B alpha and its degradation is not understood. In this report we provide evidence that phosphorylation of serine residues 32 and 36 of I kappa B alpha targets the protein to the ubiquitin-proteasome pathway. I kappa B alpha is ubiquitinated in vivo and in vitro following phosphorylation, and mutations that abolish phosphorylation and degradation of I kappa B alpha in vivo prevent ubiquitination in vitro. Ubiquitinated I kappa B alpha remains associated with NF-kappa B, and the bound I kappa B alpha is degraded by the 26S proteasome. Thus, ubiquitination provides a mechanistic link between phosphorylation and degradation of I kappa B alpha.
Activation of the transcription factor NF-kappa B is a paradigm for signal transduction through the ubiquitin-proteasome pathway: ubiquitin-dependent degradation of the transcriptional inhibitor I kappa B in response to cell stimulation. A major issue in this context is the nature of the recognition signal and the targeting enzyme involved in the proteolytic process. Here we show that following a stimulus-dependent phosphorylation, and while associated with NF-kappa B, I kappa B is targeted by a specific ubiquitin-ligase via direct recognition of the signal-dependent phosphorylation site; phosphopeptides corresponding to this site specifically inhibit ubiquitin conjugation of I kappa B and its subsequent degradation. The ligase recognition signal is functionally conserved between I kappa B alpha and I kappa B beta, and does not involve the nearby ubiquitination site. Microinjection of the inhibitory peptides into stimulated cells abolished NF-kappa B activation in response to TNF alpha and the consequent expression of E-selectin, an NF-kappa B-dependent cell-adhesion molecule. Inhibition of NF-kappa B function by specific blocking of ubiquitin ligase activity provides a novel approach for intervening in cellular processes via regulation of unique proteolytic events.
Lipopolysaccharide (LPS) is responsible for initiating host responses leading to septic shock, and tumor necrosis factor-α (TNFα) is thought to be its primary mediator. In addition, TNFα is one of the major components of the pathogenesis of insulin resistance in various conditions. It has been shown that LPS induced TNFα production in rat vascular smooth muscle cells (VSMC). However, little is known about the signaling pathway by which VSMC in culture produce TNFα. We investigated the possible signaling components involved in this pathway. LPS elicited phosphorylation of p42/44 mitogen-activated protein kinase (MAPK) and p38 MAPK, degradation of inhibitor of κB (IκB), and an increase in nuclear binding activity of activating protein-1 and nuclear factor-κB (NF-κB). Different types of NF-κB inhibitors, pyrrolidine dithiocarbamate and MG132, which specifically abolished IκB degradation and subsequent NF-κB activation by LPS, suppressed TNFα secretion from VSMC. Although PD98059, a specific MAPK kinase inhibitor and SB203580, a specific p38 MAPK inhibitor, had no effect on NF-κB activity, SB203580 suppressed TNFα secretion; however, PD98059 did not. A cotransfection assay showed that transfection of dominant negative IκB or pretreatment with SB203580 suppressed the TNFα gene promotor-dependent transcription. TNFα messenger RNA expression induced by LPS was inhibited by pyrrolidine dithiocarbamate, MG132, and SB203580, but not by PD98059. These observations indicate that TNFα production in VSMC is stimulated by LPS, and its transcription and translation are dependent on NF-κB activation through proteasome-mediated IκB degradation. It is likely that p38 MAPK may play a critical role in regulating transcription of the TNFα gene in VSMC, unlike in other cell lines.
L-2-oxothiazolidine-4-carboxylic acid (OTC) is a cysteine prodrug that maintains glutathione in tissues. Here, its effect on alcohol-induced liver injury in an enteral alcohol feeding model was investigated. Male Wistar rats were given control high-fat or ethanol containing diets enterally for 4 weeks. Treated rats received 500 mg/kg/d of dietary OTC. Ethanol delivery, weight gain, and the cyclic pattern of ethanol in the urine were not different between the OTC-ethanol and ethanol groups. After 4 weeks, serum aspartate transaminase (AST), necrosis and inflammation were elevated significantly by ethanol compared with appropriate high-fat controls, effects blocked by OTC. Moreover, ethanol elevated hepatic tumor necrosis factor alpha (TNF-alpha) messenger RNA (mRNA) and the nuclear transcription factor nuclear factor kappaB (NFkappaB) 2-3 fold. NFkappaB in isolated Kupffer cells was also increased by ethanol. These effects were all blocked by OTC treatment. Additionally, superoxide production was higher in Kupffer cells isolated from ethanol-treated rats, an effect blunted by OTC. OTC also increased circulating glutathione (GSH) levels about 2-fold; however, GSH levels were not affected by ethanol or OTC in livers from the groups studied. Surprisingly, GSH was elevated by ethanol and OTC treatment in isolated Kupffer cells about 2-fold. Moreover, GSH (Ki-10 micromol/L) and cysteinyl-glycine, but not oxidized glutathione (GSSG) or OTC, blunted the LPS-induced increase in calcium in isolated Kupffer cells, possibly by activating a glycine-gated chloride channel due to their structural similarity with glycine. Collectively, it is concluded that GSH is protective, in part, by increasing circulating GSH, which blunts activation of Kupffer cells via the glycine-gated chloride channel.
Background:
Tolerance to hemorrhagic or endotoxic shock can be induced by prior sublethal hemorrhage (SLH). The purpose of this study was to explore whether alterations in signal transduction pathways involving NF-kappaB occur in macrophages (Mphi) following induction of tolerance by SLH.
Methods:
Using a model of SLH previously shown in our lab to impart a survival benefit to subsequent hemorrhagic or endotoxic shock, rats (n = 30) were conditioned by SLH. Peritoneal Mphi were harvested 24 h after conditioning and stimulated with lipopolysaccharide (LPS) (10 microg/mL). Nuclear and cytosolic proteins were isolated 1 h later for determination of NF-kappaB activation by gel-shift assay and IkappaB-alpha by Western blot. TNF mRNA gene expression was measured 4 h after LPS stimulation by reverse transcription/polymerase chain reaction (RT/PCR). TNF protein levels were measured in cellular supernatants by enzyme-linked immunosorbent assay (ELISA) 18 h after LPS. RESULTS. LPS stimulation of sham Mphi increased NF-kappaB activation with corresponding loss of its inhibitor IkappaB-alpha. In contrast, IkappaB-alpha was not detectable following conditioning, and conditioned Mphi had NF-kappaB activation at baseline which increased minimally with LPS stimulation. LPS increased TNF gene expression and significantly increased protein production by both sham and conditioned Mphi, but this increase was greater in the sham-conditioned group.
Conclusions:
The ability of Mphi from animals made tolerant by SLH to produce TNF in vitro is conserved. Nevertheless, these same Mphi exhibit alterations in TNF gene induction and expression as well as signal transduction, specifically, changes in IkappaB-alpha and NF-kappaB activation. This suggests a role for activation of NF-kappaB in the induction of tolerance.
1. We compared the effects of calpain inhibitor I (inhibitor of the proteolysis of I kappa B and, hence, of the activation of nuclear factor kappa B (NF kappa B) and dexamethasone on (i) the circulatory failure, (ii) multiple organ dysfunction and (iii) induction of the inducible isoforms of nitric oxide (NO) synthase (iNOS) and cyclo-oxygenase (COX-2) in anaesthetized rats with endotoxic shock. 2. Injection of lipopolysaccharide (LPS, E. coli, 10 mg kg-1, i.v.) resulted in hypotension and a reduction of the pressor responses elicited by noradrenaline. This circulatory dysfunction was attenuated by pretreatment of LPS-rats with calpain inhibitor I (10 mg kg-1, i.v., 2 h before LPS) or dexamethasone (1 mg kg-1, i.v.). 3. Endotoxaemia also caused rises in the serum levels of (i) urea and creatinine (renal dysfunction), (ii) alanine aminotransferase (ALT), aspartate aminotransferase (AST) (hepatocellular injury), bilirubin and gamma-glutamyl transferase (gamma GT) (liver dysfunction), (iii) lipase (pancreatic injury) and (iv) lactate. Calpain inhibitor I and dexamethasone attenuated the liver injury, the pancreatic injury, the lactic acidosis as well as the hypoglycaemia caused by LPS. Dexamethasone, but not calpain inhibitor I, reduced the renal dysfunction caused by LPS. 4. Endotoxaemia for 6 h resulted in a substantial increase in iNOS and COX-2 protein and activity in lung and liver, which was attenuated in LPS-rats pretreated with calpain inhibitor I or dexamethasone. 5. Thus, calpain inhibitor I and dexamethasone attenuate (i) the circulatory failure, (ii) the multiple organ dysfunction (liver and pancreatic dysfunction/injury, lactic acidosis, hypoglycaemia), as well as (iii) the induction of iNOS and COX-2 protein and activity in rats with endotoxic shock. We propose that prevention of the activation of NF-kappa B in vivo may be useful in the therapy of circulatory shock or of disorders associated with local or systemic inflammation.
Nitric oxide (NO) is produced by the liver during lipopolysaccharide (LPS)-induced endotoxemia. The aim of this study was to examine whether NO, which is produced in the liver, is released from the liver surface to the intraabdominal cavity during endotoxemia. NO was quantitatively determined by chemiluminescence and a newly developed gas purge technique was used to directly measure NO released from the liver surface and the intraabdominal cavity of rats before and after LPS (0.1 mg/kg, intraperitoneally) or saline administration. The expression of inducible NO synthase (iNOS) mRNA in the liver was detected by Northern blot analysis. NO levels from both the liver surface and in the intraabdominal cavity were elevated at 2 h after LPS injection and peaked at 10 h and both the time course of NO level were well correlated with each other. Both NO levels were below the detectable range before LPS and after saline administration. Inducible NOS mRNA in the liver exhibited a sharp increase to a maximum level at 4 h after LPS injection. The present study indicates that the hepatic NO, which might have been produced by iNOS in the liver, is released from the liver surface to the intraabdominal cavity during endotoxemia.
The nuclear translocation of NF-kappa B follows the degradation of its inhibitor, I kappa B alpha, an event coupled with stimulation-dependent inhibitor phosphorylation. Prevention of the stimulation-dependent phosphorylation of I kappa B alpha, either by treating cells with various reagents or by mutagenesis of certain putative I kappa B alpha phosphorylation sites, abolishes the inducible degradation of I kappa B alpha. Yet, the mechanism coupling the stimulation-induced phosphorylation with the degradation has not been resolved. Recent reports suggest a role for the proteasome in I kappa B alpha degradation, but the mode of substrate recognition and the involvement of ubiquitin conjugation as a targeting signal have not been addressed. We show that of the two forms of I kappa B alpha recovered from stimulated cells in a complex with RelA and p50, only the newly phosphorylated form, pI kappa B alpha, is a substrate for an in vitro reconstituted ubiquitin-proteasome system. Proteolysis requires ATP, ubiquitin, a specific ubiquitin-conjugating enzyme, and other ubiquitin-proteasome components. In vivo, inducible I kappa B alpha degradation requires a functional ubiquitin-activating enzyme and is associated with the appearance of high molecular weight adducts of I kappa B alpha. Ubiquitin-mediated protein degradation may, therefore, constitute an integral step of a signal transduction process.
Nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) have been suggested to play an important role in endotoxin-mediated shock and inflammation. In this study, we investigated the effect of aqueous extract of Dichroa febrifuga Lour. (Saxifragaceae) roots, a traditional antimalarial drug, on the production of NO and TNF-alpha. The aqueous extract of D. febrifuga roots (AEDF) inhibited the secretion of NO and TNF-alpha in lipopolysaccharide (LPS) and/or interferon-gamma (IFN-gamma)-stimulated mouse peritoneal macrophages, without affecting cell viability. The protein level of inducible nitric oxide synthase (iNOS) in peritoneal macrophages was also decreased by AEDF. In addition, the serum level of NO was reduced by i.p. administration of AEDF. These results suggest that AEDF suppresses the endotoxin-induced inflammatory responses through inhibiting the production of NO and TNF-alpha, and could be used as an anti-inflammatory drug.