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Adipocyte PHLPP2 regulates HSL phosphorylation and localization a Western blots in lysates from eWAT (top) or iWAT (bottom) from Cre- or A-PHLPP2 mice fed on NCD or HFD, with quantification of various HSL phosphorylation sites (right) (n = 3 independent mice per group). b, c Western blots in lysates from differentiated PHLPP2-repressed 3T3-L1 adipocytes (b), or 3T3-L1 adipocytes expressing HA-tagged PHLPP2 (c) with quantification of p-HSL levels (right), with or without CL316,243 (n = 2 independent experiments). d Coomassie brilliant blue staining of purified human HSL (hHSL) or PHLPP2 with or without 10 μM forskolin (FSK) (top), for phosphatase assay of HSL Ser563 and Ser660 with purified PHLPP2 (bottom) (n = 2 independent experiments). e Immunofluorescence of HSL (red), Bodipy (green), and DAPI (blue) in 3T3-L1 adipocytes expressing control or HA-tagged PHLPP2 with or without CL316,243 (top). Quantification of HSL localization on lipid droplets was performed by directly counting Bodipy-positive cells from images of randomly chosen fields (bottom) (n = 3 independent samples). Scale bars, 10 μm. f, g In vitro lipolysis in differentiated control or PHLPP2-repressed adipocytes with or without CL316,243 and/or CAY10499 (f) or HSL deletion (g) (n = 3 independent mice per group). All data are shown as the means ± SEM. Statistical significance was determined by unpaired two-tailed Student’s t-test for a, e, and two-way ANOVA with Tukey’s multiple comparison test for f, g. n.s. not significant (P > 0.05).
Source publication
Increased adiposity confers risk for systemic insulin resistance and type 2 diabetes (T2D), but mechanisms underlying this pathogenic inter-organ crosstalk are incompletely understood. We find PHLPP2 (PH domain and leucine rich repeat protein phosphatase 2), recently identified as the Akt Ser473 phosphatase, to be increased in adipocytes from obese...
Citations
... Similarly, the SPATA19 gene has also been associated with mitochondrial function in the sperm line in mice [55], playing an important role in male fertility [56]. Similarly, CHST4 and TAT were linked to protein metabolism [57,58], while PHLPP2 and PDPR are involved in carbohydrate or lipid metabolic regulation [59,60]. Identifying genes or QTLs associated with traits like growth and reproduction is crucial in horse breeding, as they directly impact the breeder's economy and efficiency. ...
Analyzing genetic variability and inbreeding trends is essential for effective breed management in animal populations. To this, the characterization of runs of homozygosity (ROH) provides a good genomic approach to study the phenomena. The Polo Argentino (PA) breed, globally recognized as the best adapted to playing polo, is known for its strong influence of Thoroughbreds, intense selective breeding, and extensive use of reproductive biotechnologies. This study investigates the PA’s genomic variability, by characterizing the ROH landscape and identifying ROH islands (ROHi) as potential genomic footprints for the breed. PA horses (n = 506) were genotyped using EquineGGP™ array v5 (70 k). We calculated the inbreeding coefficient based on ROH (FROH—ancestral and recent) using a chromosomal approach. Finally, we identified genomic regions with increased ROH frequency (ROHi) and their associated genes. An average of 79.5 ROH per horse was detected, with a mean length of 4.6 Mb. The average FROH was 0.151, but most of them (54%) corresponded to ancestral inbreeding (ROH < 5.5 Mb). However, 4 ROHi were identified in ECA 1, 3, 7 and 17, containing 67 genes, some of which were related to behavior, neurodevelopment, and metabolic functions. This genomic analysis determined, for the first time, the length and location of homozygosity segments in the PA breed and identified ROHi associated with potential genomic regions and genes for positive selection in the breed.
... Fat-specific overexpression of ATGL or enhancement of HSL activity can prevent diet-induced obesity (DIO) and improve metabolic health. 44,45 Cod1, highly expressed in adipose tissue, is considered a promoter of lipolysis initiation, and its adipose tissue-specific knockout aggravated obesity and metabolic abnormalities in mice. 46 Ces1d demonstrates a direct lipolytic function by mobilizing lipid droplets upon translocation onto or proximity to them, and adipose tissue-specific knockout of Ces1d in high-fat-fed mice results in severe obesity and related metabolic disorders. ...
Background
Prediabetes, characterized by a series of metabolic abnormalities, increases the risk of diabetes and cardiovascular diseases. Tangzhiping (TZP), a clinically validated traditional Chinese medicine formula, is used to treat impaired glucose tolerance. However, the underlying mechanism of TZP in intervening prediabetes is not fully elucidated.
Purpose
The current study aimed to evaluate the protective effect of TZP against prediabetes mice and explore its potential mechanism.
Methods
After establishing a prediabetic animal model through 12 weeks of high-fat diet (HFD) feeding, mice were subjected to TZP for 8 weeks. Various parameters related to body weight, glucose and lipid metabolism, and insulin sensitivity were measured. Histopathological examinations observed adipose cell size and liver lipid deposition. The Sable Promethion system assessed energy metabolism activity. Transcriptomic analysis of Epididymal white adipose tissue (EWAT) identified enriched pathways and genes. The key genes in the enriched pathways were identified through RT-PCR.
Results
Our data revealed that the administration of TZP reduced body weight and fat mass in a prediabetes mouse model. TZP normalized the glucose and insulin levels, improved insulin resistance, and decreased plasma TC and FFA. The alleviation of adipose tissue hypertrophy and lipid deposition by TZP was demonstrated through pathological examination. Indirect calorimetry measurements indicated a potential increase in VO2 and EE levels with TZP. The results of EWAT transcription showed that TZP reversed pathways and genes related to inflammation and catabolic metabolism. RT-PCR demonstrated that the mRNA expression of inflammation and lipolysis, including Tlr2, Ccr5, Ccl9, Itgb2, Lipe, Pnpla2, Cdo1, Ces1d, Echs1, and Acad11, were changed by TZP treatment.
Conclusion
TZP effectively alleviates obesity, impaired glucose and lipid metabolism, and insulin resistance. The effect of TZP might be associated with the regulation of gene expression in dysfunctional adipose tissue.
... Pleckstrin homology domain leucine-rich repeat protein 1 phosphatase 2 (PHLPP2) in adipocytes from obese mice was higher than that in normal mice. Kim et al. [69] found that PHLPP2 levels in adipose tissue were negatively correlated with serum ADP levels in obese patients. Owing to prolonged hormone-sensitive lipase (HSL) phosphorylation, adipocyte PHLPP2 ablation causes an increase in adipose lipolysis. ...
Obesity and related metabolic syndromes have been recognized as important disease risks, in which the role of adipokines cannot be ignored. Adiponectin (ADP) is one of the key adipokines with various beneficial effects, including improving glucose and lipid metabolism, enhancing insulin sensitivity, reducing oxidative stress and inflammation, promoting ceramides degradation, and stimulating adipose tissue vascularity. Based on those, it can serve as a positive regulator in many metabolic syndromes, such as type 2 diabetes (T2D), cardiovascular diseases, non-alcoholic fatty liver disease (NAFLD), sarcopenia, neurodegenerative diseases, and certain cancers. Therefore, a promising therapeutic approach for treating various metabolic diseases may involve elevating ADP levels or activating ADP receptors. The modulation of ADP genes, multimerization, and secretion covers the main processes of ADP generation, providing a comprehensive orientation for the development of more appropriate therapeutic strategies. In order to have a deeper understanding of ADP, this paper will provide an all-encompassing review of ADP.
... Experimental studies have demonstrated that Y-BGs enhance antiinflammatory activity in macrophages through an IL-10-mediated mechanism (132). Delivering genes to specific tissues without relying on viruses can be challenging, but the application of oligopeptides in conjunction with gene-modulating compounds offers a promising solution for such cases (133)(134)(135). This fortuitous finding holds potential for precise transportation of non-viral gene-modifying technology to adipose deposits and adipose tissue macrophages, facilitating targeted outcomes (136). ...
Nonalcoholic fatty liver disease (NAFLD) and its inflammatory and often progressive subtype nonalcoholic steatohepatitis (NASH), have emerged as significant contributors to hepatic morbidity worldwide. The pathophysiology of NAFLD/NASH is multifaceted, variable, and remains incompletely understood. The pivotal role of liver-resident and recruited macrophages in the pathogenesis of NAFLD and NASH is widely acknowledged as a crucial factor in innate immunity. The remarkable plasticity of macrophages enables them to assume diverse activation and polarization states, dictated by their immunometabolism microenvironment and functional requirements. Recent studies in the field of immunometabolism have elucidated that alterations in the metabolic profile of macrophages can profoundly influence their activation state and functionality, thereby influencing various pathological processes. This review primarily focuses on elucidating the polarization and activation states of macrophages, highlighting the correlation between their metabolic characteristics and the transition from pro-inflammatory to anti-inflammatory phenotypes. Additionally, we explore the potential of targeting macrophage metabolism as a promising therapeutic approach for the management of NAFLD/NASH.
... PPARα promotes the entry of fatty acids into the mitochondria for metabolism by inducing the transcription of carnitine palmitoyltransferase 1 (CPT1) [30]. High intake of lipids inhibits the expression of PPARα, leading to lipid accumulation [31]. Therefore, PPARα activation is a potential therapeutic strategy for NAFLD, which has been examined in clinical trials [32,33]. ...
Background:
The function of flavin containing dimethylaniline monooxygenase 1 (FMO1), which is known to play a part in lipid metabolism, remains unclear in the development of nonalcoholic fatty liver disease (NAFLD). This research has the objective of examining the contributions of FMO1 in the progression of NAFLD and the associated mechanisms, particularly the peroxisome proliferator activated receptor alpha (PPARα) and ferroptosis pathways.
Methods:
An in vitro NAFLD model was established by treating L02 cells with free fatty acids (FFAs). The FMO1 and ferroptosis levels were examined in the cellular NAFLD model. FMO1 was knocked down using short-interfering RNA transfection. The effects of FMO1 knockdown on lipid accumulation, PPARα expression, and ferroptosis were examined in the cellular NAFLD model. Additionally, the effects of FMO1 and/or PPARα overexpression on lipid metabolism and ferroptosis were analyzed. Furthermore, L02 cells were pre-treated with GW7647 (PPARα agonist) or RSL3 (ferroptosis activator) and stimulated with FFAs.
Results:
The levels of FMO1 and ferroptosis were upregulated in the in vitro NAFLD model. FMO1 knockdown suppressed the FFA-induced accumulation of lipids in hepatocytes, downregulation of PPARα expression, and upregulation of ferroptosis. In contrast, FMO1 overexpression dysregulated lipid metabolism and downregulated PPARα levels. Meanwhile, PPARα overexpression mitigated the FMO1 overexpression-induced upregulation of ferroptosis and lipid accumulation. Treatment with RSL3 suppressed the effects of PPARα overexpression on lipid accumulation and FMO1 expression.
Conclusions:
FMO1 upregulates ferroptosis by suppressing PPARα in NAFLD, which leads to the dysregulation of lipid metabolism.
... Finally, Cinta Senese, Casertana, Mora Romagnola, and cosmopolitan breeds outlined ROHet islands on SSC2, SSC6, SSC14, and SSC15 that mapped genes and QTL linked to productive traits (Howard et al., 2015;Kim et al., 2021;Li et al., 2011;Qi et al., 2022;Zappaterra et al., 2021;Zhang et al., 2015). In particular, the heterozygosity-rich segment, observed on SSC6 (54.2-55.9 ...
Analysis of genomic data is becoming more and more common for the effective management of livestock breeding programmes, even in the case of local populations. In this work, the genome‐wide data of Nero Siciliano pig breed were compared to that of wild boar, Italian local and cosmopolitan breeds to investigate its genetic structure, and runs of homozygosity (ROH) and heterozygosity patterns. The Nero Siciliano has been reported to have the highest rate of genetic diversity among the Italian breeds, and a genetic variability comparable to that of the cosmopolitan breeds. Analyses of genomic structure and relationships underlined its proximity to wild boar, and an internal substructure probably linked to different family lines. The breed showed a low value of inbreeding estimated from ROH, and the highest diversity index among the Italian breeds, even if lower than that of the cosmopolitans. Four ROH islands in three chromosomes (SSC8, SSC11, and SSC14) and one heterozygosity‐rich region (SSC1) were identified in Nero Siciliano, highlighting genomic regions related to productive QTL. Across breeds, SSC8 and SSC14 were the chromosomes with most ROH islands, with Mora Romagnola and wild boar showing the highest level of autozygosity. Chromosomes SSC2, SSC6, SSC8 and SSC13 showed the majority of runs of heterozygosity regions, mainly found in the cosmopolitan pig breeds, which reported several genes associated with health‐related QTL. The outlined results can help to better identify the genomic profile of this local breed in order to plan matings, maintain adequate internal diversity and exploit the production system.
... Pleckstrin homology domain and leucine-rich repeat protein phosphatase 2 (PHLPP2) is a protein phosphatase that mediates the dephosphorylation of various substrates and maintains cellular homeostasis [6][7][8][9]. The dysregulation of PHLPP2 is linked to multiple diseases, including metabolic disease, cardiovascular disease, and cancer [10][11][12][13][14][15]. PHLPP2 can dephosphorylate and inactivate the kinase Akt, thereby participating in diverse cellular processes [6,16]. ...
Objective
Pleckstrin homology domain and leucine-rich repeat protein phosphatase 2 (PHLPP2) is linked to various pathological states. However, whether PHLPP2 mediates diabetic retinopathy is unaddressed. This work explored the biological function of PHLPP2 in modulating high glucose (HG)-elicited damage of retinal ganglion cells (RGCs), an in vitro model for studying diabetic retinopathy.Methods
Mouse RGCs were treated with HG to establish a cell model. PHLPP2 was silenced by transfecting specific shRNAs targeting PHLPP2. RT-qPCR, immunoblotting, CCK-8 assay, flow cytometry, TUNEL assay, and ELISA were carried out.ResultsSignificant increases in PHLPP2 levels were observed in cultured RGCs exposed to HG. The severe damages evoked by HG to RGCs were remarkably weakened in PHLPP2-silenced RGCs, including improved cell survival, attenuated cell apoptosis, repressed oxidative stress, and prohibited proinflammatory response. The silencing of PHLPP2 strengthened the activation of Nrf2 in HG-treated RGCs via modulation of the Akt–GSK–3β axis. Interruption of the Akt–GSK–3β axis reversed PHLPP2-silencing-elicited Nrf2 activation. The protective effects of PHLPP2 silencing on HG-induced injury of RGCs were diminished by Nrf2 inhibition.Conclusions
The loss of PHLPP2 was beneficial for HG-injured RGCs through the effect on the Akt–GSK–3β–Nrf2 pathway. This work suggests a possible role of PHLPP2 in diabetic retinopathy.
... However, both the studies negate the involvement of PHLPP2 in insulin resistance associated with obesity or diabetes. Recently, only one study reported higher adipocyte PHLPP2 levels in obese mice as compared to lean mice and a possible inter-organ cross-talk with the liver, indicating a role of PHLPP2 in obesity-related metabolic disorders [11]. In the liver, aged and obese mice also display lower PHLPP2 protein levels, affecting Akt signaling and inducing obesity-induced insulin resistance [12]. ...
... Interestingly, these studies negate any involvement of PHLPP2 under pathological disorders [9,10]. Wu et al. and Kim et al. reported elevated PHLPP2 from adipose tissue of obese leptin receptor-deficient db/db mice and HFD fed mice respectively [11,33]. The study by Kim et al. comparing C57BL/6 J mice, obese db/db mice to liver-specific PHLPP2 knockout mice under normal chow or high fat diet reported PHLPP2 knockout induced lipogenesis [34]. ...
Background
The aim of the present study was to determine the role of individual PHLPP isoforms in insulin signaling and insulin resistance in neuronal cells.
Methods
PHLPP isoforms were either silenced or overexpressed individually, and the effects were observed on individual Akt isoforms, AS160 and on neuronal glucose uptake, under insulin sensitive and resistant conditions. To determine PHLPP regulation itself, we tested effect of scaffold protein, Scribble, on PHLPP isoforms and neuronal glucose uptake.
Results
We observed elevated expression of both PHLPP1 and PHLPP2 in insulin resistant neuronal cells (Neuro-2A, mouse neuroblastoma; SHSY-5Y, human neuroblastoma) as well as in the whole brain lysates of high-fat-diet mediated diabetic mice. In insulin sensitive condition, PHLPP isoforms differentially affected activation of all Akt isoforms, wherein PHLPP1 regulated serine phosphorylation of Akt2 and Akt3, while PHLPP2 regulated Akt1 and Akt3. This PHLPP mediated Akt isoform specific regulation activated AS160 affecting glucose uptake. Under insulin resistant condition, a similar trend of results were observed in Akt isoforms, AS160 and glucose uptake. Over-expressed PHLPP isoforms combined with elevated endogenous expression under insulin resistant condition drastically affected downstream signaling, reducing neuronal glucose uptake. No compensation was observed amongst PHLPP isoforms under all conditions tested, indicating independent roles and pointing towards possible scaffolding interactions behind isoform specificity. Silencing of Scribble, a scaffolding protein known to interact with PHLPP, affected cellular localization of both PHLPP1 and PHLPP2, and caused increase in glucose uptake.
Conclusions
PHLPP isoforms play independent roles via Scribble in regulating Akt isoforms differentially, affecting AS160 and neuronal glucose uptake.
... Mice with adipose-tissue-specific knockout of ATGL had increased adiposity, defective cold adaptation and conversion of BAT into a WAT-like tissue 14 . On the contrary, fat-specific overexpression of ATGL or enhancement of HSL activity prevented diet-induced obesity (DIO) and improved metabolic health 15,16 . The aforementioned evidence indicates the crucial role of the key lipolytic enzymes in maintaining energy homeostasis and preventing obesity. ...
... Global vanin-1 deficiency, adipose-specific knockout of sirtuin6 (Sirt6) or knockout of HuR in adipose tissue inhibited ATGL expression and lipolysis, which exacerbated DIO and insulin resistance in mice 34,35,41 . Boosting adipose HSL-mediated lipolytic activity by adipocyte-specific ablation of PH domain and leucine rich repeat protein phosphatase 2 (PHLPP2) reduced DIO, improved whole-body glucose tolerance and ameliorated fatty liver in mice 16 . Consistent with the aforementioned studies, our study demonstrated that Cdo1-mediated expression of ATGL and HSL and subsequent lipolysis played an important part in ameliorating obesity and related metabolic disorders. ...
Cysteine dioxygenase 1 (Cdo1) is a key enzyme in taurine synthesis. Here we show that Cdo1 promotes lipolysis in adipose tissue. Adipose-specific knockout of Cdo1 in mice impairs energy expenditure, cold tolerance and lipolysis, exacerbates diet-induced obesity (DIO) and decreases adipose expression of the key lipolytic genes encoding ATGL and HSL, with little effect on adipose taurine levels. White-adipose-specific overexpression of ATGL and HSL blunts the role of adipose Cdo1 deficiency in promoting DIO. Mechanistically, Cdo1 interacts with PPARγ and facilitates the recruitment of Med24, the core subunit of mediator complex, to ATGL and HSL gene promoters, thereby transactivating their expression. Further, mice with transgenic overexpression of Cdo1 show better cold tolerance, ameliorated DIO and higher lipolysis capacity. Thus, we uncover an unexpected and important role of Cdo1 in regulating adipose lipolysis.
... Accordingly, in the last 20 years, various HSL inhibitors, including natural and synthetic products, have been described for the treatment of diabetes and lipid disorders [63]. Moreover, the use of inhibitors and activators of HSL activity is indispensable in basic research for deciphering the HSL's role in various pathophysiological conditions [112][113][114][115]. The only specific, reversible non-competitive inhibitor of HSL is BAY 59-9435 (BAY), a derivative of 5-(2H)-isoxazolonyl urea. ...
Since the 1950s, one of the goals of adipose tissue research has been to determine lipolytic and lipogenic activity as the primary metabolic pathways affecting adipocyte health and size and thus representing potential therapeutic targets for the treatment of obesity and associated diseases. Nowadays, there is a relatively large number of methods to measure the activity of these pathways and involved enzymes, but their applicability to different biological samples is variable. Here, we review the characteristics of mean lipogenic and lipolytic enzymes, their inhibitors, and available methodologies for assessing their activity, and comment on the advantages and disadvantages of these methodologies and their applicability in vivo, ex vivo, and in vitro, i.e., in cells, organs and their respective extracts, with the emphasis on adipocytes and adipose tissue.
