Inhibited Lipid Accumulation Research Articles

Ebastine-mediated destabilization of E3 ligase MKRN1 protects against metabolic dysfunction-associated steatohepatitis

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a chronic condition encompassing metabolic dysfunction-associated steatotic liver (MASL) and metabolic dysfunction-associated steatohepatitis (MASH), which can progress to fibrosis, cirrhosis, or hepatocellular carcinoma (HCC). The heterogeneous and complex nature of MASLD complicates optimal drug development. Ebastine, an antihistamine, exhibits antitumor activity in various types of cancer. However, its effects on MASH remain unexplored. In the present study, we identified ebastine as a potential treatment for MASH. Our results indicated that ebastine acts as a novel MKRN1 inhibitor by promoting MKRN1 destabilization through self-ubiquitination, leading to AMP-activated protein kinase (AMPK) activation. Ebastine appeared to bind to the C-terminal domain of MKRN1, particularly at residues R298 and K360. Notably, Mkrn1 knockout (KO) mice demonstrated resistance to MASH, including obesity, steatosis, inflammation, and fibrosis under high-fat-high-fructose diet (HFHFD) conditions. Additionally, liver-specific Mkrn1 knockdown using AAV8 alleviated MASH symptoms in HFHFD-fed mice, implicating MKRN1 as a potential therapeutic target. Consistent with these findings, treatment with ebastine significantly reduced the risk of MASH in HFHFD-fed mice, with a decrease in MKRN1 expression and an increase in AMPK activity. Our study suggests that ebastine binds to MKRN1, promoting its destabilization and subsequent degradation by stimulating its ubiquitination. This enhances AMPK stability and activity, suppressing lipid accumulation, inflammation, and fibrosis. Moreover, the knockout of Mkrn1 mice decreased the risk of MASH, suggesting that ebastine could be a promising therapeutic agent for the treatment of MASH.

Quercetin-Driven Akkermansia Muciniphila Alleviates Obesity by Modulating Bile Acid Metabolism via an ILA/m6A/CYP8B1 Signaling.

Global health is increasingly challenged by the growing prevalence of obesity and its associated complications. Quercetin, one of the most important dietary flavonoids, is being explored as an effective therapy for obesity with its mechanism remains understudied. Here in this study, it is demonstrated that quercetin intervention significantly reverses obesity-related phenotypes through reshaping the overall structure of microbiota, especially boosting colonization of the beneficial gut commensal Akkermansia muciniphila (A. muciniphila). Enrichment of A. muciniphila leads to generate more indole-3-lactic acid (ILA) to upregulate the expression of 12α-hydroxylase (CYP8B1) via fat mass and obesity-associated protein (FTO)/ N6-methyladenosine (m6A)/YTHDF2 manner, thereby facilitating cholesterol converts to cholic acid (CA). CA in turn drastically suppresses lipid accumulation via activating the farnesoid X receptor (FXR) in adipose tissue. This work introduces a novel therapeutic target for addressing obesity and expands upon the current limited understanding of the mediator function of m6A modifications in microorganism-influenced bile acid (BA) metabolism.

Knockdown of FAS2 Impairs Fecundity by Inhibiting Lipid Accumulation and Increasing Glycogen Storage in Locusta migratoria

Fatty acid synthase (FAS) is a pivotal gene in the lipid synthesis pathway and plays a crucial role in insect energy metabolism. Locusta migratoria, as one of the major agricultural pests, requires identification of new targets to control or reduce its reproductive capacity for effective locust pest management strategies. In this study, we focused on L. migratoria and identified FAS2 as a potential target gene with functional significance in lipid metabolism and reproduction based on sequence characteristics analysis and tissue-expression patterns of five FAS genes. Subsequently, through RNA interference (RNAi) targeting FAS2 expression, we assessed alterations in lipid and carbohydrate metabolism-related gene expression levels, lipid and carbohydrate contents, ovarian development, and reproductive capacity using experimental techniques such as RT-qPCR, ELISA, and morphological observations. Our findings revealed that interference with FAS2 upregulated genes involved in lipid degradation, including Lsd-1, Lsd-2, Lipase3, and Brummer, while significantly decreasing the TAG content and fat accumulation. At the level of carbohydrate metabolism, FAS2 silencing led to significant upregulation of key genes TPS and GS in the synthesis pathway, resulting in increased glycogen and trehalose content. In addition, FAS2 interference resulted in a significant reduction of Vg mRNA expression level sand protein content in L. migratoria, followed by delayed ovarian development and reduced egg production. This further confirms that impaired FAS2 function prompts L. migratoria to enhance lipid degradation and sugar storage to maintain the energy balance, while reducing the energy investment into reproduction. Collectively, the results of this study suggest that FAS2 can serve as a novel molecular target for controlling L. migratoria.

A Mechanistic Insight into the Effects of Marine Sources as Functional Foods Against Metabolic Syndrome: A Review Article

Introduction: Marine organisms are a rich source of metabolites that can be considered functional foods and produce new medicines against metabolic syndrome, including hypertension, dyslipidemia, hyperglycemia, obesity, insulin resistance, and hyperinsulinemia. Method: A total of 609 publications were searching results, 283 were screened entirely, and 43 duplications were removed. Results: These compounds, found in marine organisms such as algae, sponges, and mollusks, have vasodilatory properties, inhibit certain enzymes involved in blood pressure regulation, and exhibit anti-oxidant activity, making them promising candidates for developing hypertension treatments. Marine organisms showed an improvement in insulin sensitivity and blood glucose level regulation, suggesting their potential in developing novel therapies for diabetes management. Furthermore, some marine sources inhibit lipid accumulation, regulate metabolism, and suppress appetite, indicating their potential as therapeutic agents in combating obesity. Conclusion: However, further studies on the pharmacological effects of marines should be performed to evaluate the potential of these compounds to produce novel drugs, but they hope this study paves and lights the way.

Japanese Leaf Burdock Extract Inhibits Adipocyte Differentiation in 3T3-L1 Cells.

Burdock, Arctium lappa Linn. (Asteraceae), is cultivated in East Asian for its edible roots, and its seeds are used in a herbal medicine. Burdock seeds and roots exhibit anti-adipogenic activity. However, the anti-adipogenic activity of the aboveground parts of burdock remains underexplored. Therefore, this study determined the anti-adipogenic effects of burdock leaf extract on 3T3-L1 adipocytes. Seventy percent ethanol (EtOH) extract of burdock leaves, which inhibited lipid accumulation, was fractionated into five fractions using Diaion® HP-20. EtOH eluted fractions (40 and 80%) strongly inhibited lipid accumulation. A common compound in these fractions was onopordopicrin (OPP), and purified OPP suppressed lipid accumulation and inhibited adipocyte differentiation. α, β-unsaturated carbonyl structure of OPP suggests potential electrophilic reactivity with polyfunctional thiol-trapping agents like cysteine residues. Indeed, the anti-adipogenic effects of OPP disappeared with the addition of cysteamine, which possesses a thiol group. Rhodamine-maleimide assay showed that OPP binds to the cysteine residues of adipocyte-specific transcription factor, peroxisome proliferator-activated receptor γ (PPARγ). Hence, our findings suggest that OPP exerts its anti-adipogenic action via binding to cysteine residues in signal proteins like PPARγ, inhibiting their activation. Thus, burdock leaf extract is a potential source of OPP, a bioactive compound with anti-obesity activity.

Phyllanthus emblica Prevents Adipogenesis by Regulating Histone Acetylation.

Phyllanthus emblica is widely used in Ayurvedic preparations against multiple disorders and contains various bioactive components. This study aimed to determine the preventive effect of P. emblica on obesity by evaluating the inhibition of adipogenesis and the related regulatory epigenetic mechanisms during 3T3-L1 differentiation. The ethyl acetate fraction of P. emblica (EFPE) effectively inhibited lipid accumulation and triglyceride (TG) production in 3T3-L1 adipocytes. It also inhibited histone acetyltransferase (HAT) activity and regulated Pcaf-specific H3K9 acetylation and the expression of adipogenesis-related genes during adipocyte differentiation. Phenolic compounds were the main components of EFPE, of which gallic acid (GA) exhibited the strongest inhibitory effect on lipid accumulation and TG production. Notably, GA effectively regulated adipogenesis-mediated gene expression through H3K9 acetylation. These findings, along with the experiment results, suggest that EFPE containing GA is a potent agent for preventing obesity by regulating H3K9 acetylation.

Pancreatic lipase inhibitory activity and hypolipidaemic effects of the pomace of Gamazumi (Viburnum dilatatum) fruit in mice with high-fat diet-induced obesity

Abstract Obesity prevention is emphasised as the first step in preventing lifestyle-related diseases. Inhibition of pancreatic lipase decreases the absorption of fat and prevents obesity, but synthesised inhibitor has side effects, such as diarrhoea. Viburnum dilatatum (V. dilatatum) fruits are cultivated and squeezed to manufacture beverages, and the pomace is generated. Therefore, the pancreatic lipase inhibitory and hypolipidaemic effects of V. dilatatum pomace in mice with high-fat diet-induced obesity were investigated to identify natural inhibitors for obesity prevention. The pancreatic lipase-inhibitory ingredients in the pomace were identified as quercetin and kaempferol using high-performance liquid chromatography and ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. In mice fed with high-fat diets for 30 days, the ingestion of V. dilatatum pomace powder or its lipase inhibitory fraction led to decrease adipose tissue weights and plasma triacylglycerol concentration. After oral administration of soybean oil, postprandial hyperlipidaemia was suppressed in mice fed with a fraction diet. Moreover, their faecal lipid excretion was greater than that of the control mice, demonstrating that the hypolipidaemic effect of V. dilatatum pomace was induced by the pancreatic lipase inhibitory activity of quercetin and kaempferol. These results suggest that V. dilatatum pomace suppresses lipid accumulation and is useful to prevent obesity.

ATGL regulates renal fibrosis by reprogramming lipid metabolism during the transition from AKI to CKD.

Acute kidney injury (AKI) can progress to chronic kidney disease (CKD) and subsequently to renal fibrosis. Poor repair of renal tubular epithelial cells (TECs) after injury is the main cause of renal fibrosis. Studies have shown that restoring damaged fatty acid β-oxidation (FAO) can reduce renal fibrosis. Adipose triglyceride lipase (ATGL) is a key enzyme that regulates lipid hydrolysis. This study, for the first time, demonstrated that ATGL was downregulated in the renal TEC in the AKI-CKD transition mouse model. Moreover, treatment with the ATGL inhibitor atglistatin exacerbated lipid accumulation and downregulated the FAO level and mitochondrial function, while it increased the level of oxidative stress injury and apoptosis, resulting in aggravated renal fibrosis. In contrast, ATGL overexpression suppressed lipid accumulation, improved the FAO level and mitochondrial function, and attenuated oxidative stress and apoptosis, thereby ameliorating fibrosis invitro and invivo. In summary, ATGL regulates renal fibrosis by reprogramming lipid metabolism in renal TECs. This study provided new avenues and targets for treating CKD.

High-density lipoprotein alleviates ocular inflammation by downregulating M1 microglia and pyroptosis through regulating lipid accumulation and Caveolin-1 expression.

Uveitis encompasses a group of intraocular inflammatory diseases that are often associated with low levels of high-density lipoprotein (HDL). The role of HDL in intraocular inflammatory diseases remains unclear. In our research, we established an endotoxin-induced uveitis (EIU) model to investigate the role of HDL. Our study indicated that HDL could suppress ocular inflammation and restore retinal function in EIU mice. Specifically, HDL intervention effectively inhibited microglial activation and promoted the transformation of microglia from the M1 phenotype to the M2 phenotype. Furthermore, HDL intervention reduced microglial pyroptosis. Additionally, HDL was found to inhibit lipid accumulation in LPS-induced microglia, which is associated with inflammation, M1 polarization, and pyroptosis, by enhancing the expression of Caveolin-1 (CAV-1). Finally, we demonstrated that the function of HDL may be partially dependent on CAV-1 expression. We conclude that HDL inhibits pathological ocular inflammation by regulating M1/M2 phenotype polarization and pyroptosis through the modulation of lipid accumulation and CAV-1 expression. This suggests that HDL may represent a novel therapeutic strategy for ocular inflammation.

Bacillus lipopeptides inhibit lipase activity and promote 3T3-L1 preadipocyte differentiation

Bacillus lipopeptides have been reported to display anti-obesity effects. In the present study, Lipopeptides from Bacillus velezensis FJAT-45028 that consisted of iturin, fengycin and surfactin were reported. The lipopeptides exhibited a strong lipase inhibition activity in a concentration-dependent manner with a half maximal inhibitory concentration of 0.012 mg/mL, and the inhibition mechanism and type were reversible and competitive, respectively. Results of CCK8 assay showed that 3T3-L1 preadipocyte cells were completely viable under treatment of 0.050–0.2 mg/mL lipopeptides for 24 or 48 h. It was found that the lipopeptides could increase lipid droplets in the differentiated 3T3-L1 adipocytes in tested concentration and suppress the expression of peroxisome proliferator-activated receptor gamma (PPARγ). These results indicated the potential anti-obesity mechanism of the tested lipopeptides might be to inhibit lipase activity but not to suppress lipid accumulation in the adipocytes. Moreover, the lipopeptides could elevate glucose utilisation by 14.43%–33.81% in the differentiated 3T3-L1 adipocytes.

Gamma-aminobutyric acid fermentation and its fermented extracts on α-glucosidase inhibition and anti-obesity effect.

Levilactobacillus brevis KCL010 was fermented in a simple medium containing 8% (w/v) of rice bran extract. We modified the carbon, nitrogen, and initial pH conditions using 10g/L of sucrose, 10g/L of yeast extract, and 5.0 of pH, respectively. To minimize the pH increase due to decarboxylation, we fermented 100mL of modified synthetic medium containing citrate-phosphate buffer (CPB, pH 5.0) of 25-200mM in 250mL Erlenmeyer flasks. After 72h of fermentation with 50mM CPB, the maximum GABA concentration and conversion efficiency were 3.42g/L and 22.39%. Furthermore, the potential α-glucosidase inhibitory activity, MTT assay, and oil red O staining were determined by fermented extracts of L. brevis KCL010. At the highest concentration of 500μg/mL, the α-glucosidase inhibition percentages for non-fermented rice bran (NFRB), rice bran fermented by L. brevis (RBFL), and GABA (analytical standard) extracts were 55.03%, 58.37%, and 59.48%, respectively. All extracts exceeded 80% viability, suggesting that there was no cytotoxic to 3T3-L1 adipocytes. The rice bran fermented by L. brevis (RBFL) extract shows a high inhibition of lipid accumulation by 29.33% compared to those of extracts.

Statement of Retraction: The myocardial infarction-associated transcript 2 inhibits lipid accumulation and promotes cholesterol efflux in oxidized low-density lipoprotein-induced THP-1-derived macrophages via inhibiting mitogen-activated protein kinase signaling and activating the nuclear factor erythroid-related factor 2 signaling pathway

Statement of Retraction: The myocardial infarction-associated transcript 2 inhibits lipid accumulation and promotes cholesterol efflux in oxidized low-density lipoprotein-induced THP-1-derived macrophages via inhibiting mitogen-activated protein kinase signaling and activating the nuclear factor erythroid-related factor 2 signaling pathway

Indole-3-Carboxaldehyde Inhibits Inflammatory Response and Lipid Accumulation in Macrophages Through the miR-1271-5p/HDAC9 Pathway.

Indole-3-carboxaldehyde (ICA), a microbiota-derived tryptophan metabolite, has been reported to protect against atherosclerosis. However, the molecular mechanisms for its atheroprotective effect remain largely unknown. This study aimed to explore the influence of ICA on lipid accumulation and inflammatory response in THP-1 macrophage-derived foam cells. Our results showed that administration of ICA upregulated the expression of miR-1271-5p, ATP binding cassette transporter A1 (ABCA1) and ABCG1, downregulated histone deacetylase 9 (HDAC9) expression and inhibited macrophage lipid accumulation. ICA treatment also facilitated macrophage polarisation to the M2 phenotype and alleviated inflammatory response, as evidenced by decreased IL-6 levels and increased IL-10 levels. HDAC9 was identified as a direct target of miR-1271-5p. HDAC9 overexpression or miR-1271-5p knockdown decreased the effect of ICA on ABCA1 and ABCG1 expression as well as inflammatory response. Taken together, these results suggest that ICA can suppress lipid accumulation and mitigate inflammatory response in macrophages by activating the miR-1271-5p/HDAC9 signalling cascade, thereby providing new explanations for how ICA reduces atherosclerosis.

Meteorin-like protein alleviates intervertebral disc degeneration by suppressing lipid accumulation in nucleus pulposus cells via PPARα-CPT1A activation.

Disturbances in lipid metabolism are closely related to intervertebral disc degeneration (IDD). However, the lipid metabolism characteristics of nucleus pulposus (NP) cells during IDD are unclear. Exercise protects against IDD and acts as a potent mediator of organ metabolism, in which muscle-secreted myokines actively participate. However, whether exercise-induced myokines alleviate IDD by regulating lipid metabolism in NP cells remains unknown. The present study revealed that lipid accumulation is the metabolic reprogramming phenotype in NP cells during IDD, which was attributed to an imbalance between increased fatty acid/triglyceride synthesis and diminished utilization, and was further associated with extracellular matrix (ECM) degradation and cell senescence. To explore the interaction between exercise and IDD, Sprague-Dawley rats were subjected to five weeks of treadmill running exercise, and rats in the exercise group exhibited less severe IDD than did those in the sedentary group. The expression of meteorin-like protein (Metrnl), a newly-discovered myokine that participates in lipid metabolism regulation, was observed to increase in muscle, serum and NP tissue after exercise. Moreover, Metrnl ameliorated lipid accumulation in NP cells and further alleviated ECM degradation and cell senescence. Mechanistically, Metrnl activated the fatty acid β-oxidation rate-limiting enzyme carnitine palmitoyltransferase 1A (CPT1A) via peroxisome proliferator-activated receptor α (PPARα) to increase lipid utilization in NP cells. This study provides insight into the lipid metabolic features of NP cells in IDD and reveals the intrinsic connections among exercise, metabolism and IDD, with the myokine Metrnl emerging as a pivotal mediator with therapeutic potential.

Betaine Suppresses Lipid Accumulation in Liver: Inhibition of FoxO6 and PPARγ Interaction.

Betaine is the major water-soluble component of Lycium chinensis. Although there are reports of a protective effect of betaine on fatty liver disease, the underlying mechanisms are unclear. We attempted to elucidate the molecular regulation of betaine on hyperglycemia-induced hepatic lipid accumulation via Forkhead box O (FoxO)6 activation. HepG2 cells and liver tissue isolated from db/db mice treated with betaine were used. The present study investigated whether betaine ameliorates hepatic steatosis by inhibiting FoxO6/peroxisome proliferator-activated receptor gamma (PPARγ) signaling in liver cells. Interestingly, betaine notably decreased lipid accumulation in tissues with FoxO6-induced mRNA expression of lipogenesis-related genes. Furthermore, betaine inhibited the FoxO6 interaction with PPARγ and cellular triglycerides in high-glucose- or FoxO6-overexpression-treated liver cells. In addition, we confirmed that betaine administration via oral gavage significantly ameliorated hepatic steatosis in db/db mice. We conclude that betaine ameliorates hepatic steatosis, at least in part, by inhibiting the interaction between FoxO6 and PPARγ, thereby suppressing lipogenic gene transcription.

Intrahepatic levels of microbiome-derived hippurate associates with improved metabolic dysfunction-associated steatotic liver disease.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is characterised by lipid accumulation in the liver and is often associated with obesity and type 2 diabetes. The gut microbiome recently emerged as a significant player in liver metabolism and health. Hippurate, a host-microbial co-metabolite has been associated with human gut microbial gene richness and with metabolic health. However, its role on liver metabolism and homeostasis is poorly understood. We characterised liver biospies from 318 patients with obesity using RNAseq and metabolomics in liver and plasma to derive associations among hepatic hippurate, hepatic gene expression and MASLD and phenotypes. To test a potential beneficial role for hippurate in hepatic insulin resistance, we profile the metabolome of (IHH) using ultra-high-performance liquid chromatography coupled to high-resolution tandem mass spectrometry (UHPLC-MS/MS), and characterised intracellular triglyceride accumulation and glucose internalisation after a 24 h insulin exposure. We first report significant associations among MASLD traits, plasma and hepatic hippurate. Further analysis of the hepatic transcriptome shows that liver and plasma hippurate are inversely associated with MASLD, implicating lipid metabolism and regulation of inflammatory responses pathways. Hippurate treatment inhibits lipid accumulation and rescues insulin resistance induced by 24-hour chronic insulin in IHH. Hippurate also improves hepatocyte metabolic profiles by increasing the abundance of metabolites involved in energy homeostasis that are depleted by chronic insulin treatment while decreasing those involved in inflammation. Altogether, our results further highlight hippurate as a mechanistic marker of metabolic health, by its ability to improve metabolic homeostasis as a postbiotic candidate.

Isolation and purification of polyphenols, hypoglycemic and hypolipidemic and active constituent analysis of walnut septum polyphenols

Walnut septum is rich in polyphenolic substances, but the hypoglycaemic and hypolipidemic effects of walnut septum polyphenols (WSP) and the main active compounds responsible for these effects are still unknown. In the present study, WSP was firstly extracted, and then purified using D101 macroporous resin, and the purity of WSP obtained was 87.36 %. WSP inhibited the activity of α-glucosidase and pancreatic lipase with IC50 values of 36.2 μg/mL and 155.9 μg/mL, respectively. Furthermore, WSP increased glucose consumption and decreased oxidative stress in insulin-resistant C2C12 cells, as well as inhibited lipid accumulation in 3 T3-L1 adipocytes. Eight naturally active ligands for α-glucosidase and pancreatic lipase were identified from WSP by affinity ultrafiltration combined with liquid chromatography-mass spectrometry analytical methods. Further molecular docking showed strong binding between these eight ligands and the enzyme. The results of the study lay a scientific foundation for the development of walnut septum as a functional food for hypoglycemic and hypolipidemic.

Ameliorative potential of ellagic acid via PPARγ against hyperlipidemia: Insights from mice and zebrafish

Peroxisome proliferator-activated receptor γ (PPARγ) was a major transcription factor and performed an essential function in the regulation of lipid metabolism. Ellagic acid (EA) was a natural phytochemical extracted from berries and pomegranates, and research had demonstrated its ameliorative effects on lipid metabolism disorders. We intended to explore the interventional effect and mechanism of action of EA on hyperlipidemia. In vitro experiments revealed that EA could reduce lipid accumulation in HepG2 cells. EA was found to bind stably to PPARγ, suppressing its nuclear translocation as well as its luciferase activity. It could also down-regulate the expression of genes and proteins related to lipid metabolism upstream and downstream of PPARγ. To explore the impact of EA on hyperlipidemia in vivo from different levels and perspectives, we established hyperlipidemia mice and hyperlipidemia zebrafish models. Consistent with the in vitro results, in vivo experiments revealed that EA attenuated lipid accumulation in hyperlipidemia mice and reduced the levels of related inflammatory factors in their serum. EA also inhibited lipid accumulation by decreasing the expression of upstream and downstream lipid-associated genes of pparg in hyperlipidemia zebrafish. In conclusion, EA could effectively improve hyperlipidemia by regulating PPARγ.

Coptisine inhibits lipid accumulation in high glucose- and palmitic acid-induced HK-2 cells by regulating the AMPK/ACC/CPT-1 signaling pathway.

AMPK (Adenosine 5'-Monophosphate activated Protein Kinase) functions as a fundamental regulator of glycolipid metabolism by regulating the rate-limiting enzyme activity of ACC (Acetyl-CoA Carboxylase, essential for fatty acid biosynthesis) and CPT-1 (Carnitine palmitoyltransferase-1, essential for mitochondrial fatty acid oxidation, FAO) in cells, which is crucial for maintaining energy homeostasis in the human body. Coptisine (COP) is a natural berberine and isoquinoline alkaloid in Coptis chinensis that has been used as a traditional Chinese herb to treat diabetes for thousands of years, but its mechanism of action is still unclear. In this study, we investigated the anti-lipid accumulation effect and mechanism of COP in high glucose and palmitic acid-induced HK-2 cells. Compared with the control HK-2 cells, the model HK-2 cells exhibited markedly greater lipid deposition, after treatment with high glucose (HG, 30 mM) and palmitic acid (PA, 250 µM) for 24h. However, COP significantly decreased the TC and TG levels in a dose dependent manner (2.5, 5, and 10 µM). Moreover, COP dramatically enhanced the effect of the positive control (AICAR, Acadesine, an AMPK activator) in alleviating lipid deposition, which was reversed by the negative control (Compound C, an AMPK inhibitor). Furthermore, COP also increased p-AMPK, p-ACC and CPT-1 protein expression. Our results indicate that COP can effectively protects HK-2 cells against HG- and PA-induced lipid accumulation by affecting the AMPK/ACC/CPT-1 signaling pathway, inhibiting de novo lipogenesis and enhancing the FAO processes, which offers novel insights for the application of COP in the clinic.

Molecular Insights into the Inhibition of Lipid Accumulation in Hepatocytes by Unique Extracts of Ashwagandha.

We investigated the effect of purified withanolides and extracts derived from Ashwagandha on steatosis, the abnormal accumulation of fat that can lead to non-alcoholic fatty liver disease (NAFLD). Collaborator of ARF (CARF, also known as CDKN2AIP, a protein that regulates hepatic lipid metabolism, fat buildup, and liver damage) was used as an indicator. Six withanolides (Withaferin A, Withanone, Withanolide B, Withanoside IV, Withanoside V, and Withanostraminolide-12 deoxy) reversed the decrease in CARF caused by exposure to free fatty acids (FFAs) in liver-derived cells (HepG2 hepatocytes). After analyzing the effects of these withanolides on CARF mRNA and protein levels, FFA accumulation, protein aggregation, and oxidative and DNA damage stresses, we selected Withaferin A and Withanone for molecular analyses. Using the palmitic-acid-induced fatty acid accumulation stress model in Huh7 cells, we found a significant reduction in the activity of the key regulators of lipogenesis pathways, including sterol regulatory element-binding protein-1c (SREBP-1c), fatty acid synthase (FASN), and peroxisome proliferator-activated receptors (PPARγ and PPARα). This in vitro study suggests that low, non-toxic doses of Withaferin A, Withanone, or Ashwagandha extracts containing these withanolides possess anti-steatosis and antioxidative-stress properties. Further in vivo and clinical studies are required to investigate the therapeutic potential of these Ashwagandha-derived bioactive ingredients for NAFLD.