BACKGROUND Chronic hepatitis B virus(HBV)infection is often associated with increased lipid deposition in hepatocytes.However,when combined with non-alcoholic fatty liver disease or hyperlipidemia,it tends to have a l...BACKGROUND Chronic hepatitis B virus(HBV)infection is often associated with increased lipid deposition in hepatocytes.However,when combined with non-alcoholic fatty liver disease or hyperlipidemia,it tends to have a lower HBV deoxyribonucleic acid(DNA)load.The relationship between lipid metabolism and HBV DNA replication and its underlying mechanisms are not well understood.AIM To investigate the relationship between lipid metabolism and HBV DNA replication and its underlying mechanisms.METHODS 1603 HBsAg-seropositive patients were included in the study.We first explored the relationship between patients'lipid levels,hepatic steatosis,and HBV DNA load.Also,we constructed an HBV infection combined with a hepatic steatosis cell model in vitro by fatty acid stimulation of HepG2.2.15 cells to validate the effect of lipid metabolism on HBV DNA replication in vitro.By knocking down and overexpressing Plin2,we observed whether Plin2 regulates autophagy and HBV replication.By inhibiting both Plin2 and cellular autophagy under high lipid stimulation,we examined whether the Plin2-autophagy pathway regulates HBV replication.RESULTS The results revealed that serum triglyceride levels,high-density lipoprotein levels,and hepatic steatosis ratio were significantly lower in the HBV-DNA high load group.Logistic regression analysis indicated that hepatic steatosis and serum triglyceride levels were negatively correlated with HBV-DNA load.Stratified analysis by HBeAg showed significant negative correlations between HBV-DNA load and hepatic steatosis ratio in both HBeAgpositive and HBeAg-negative groups.An in vitro cell model was developed by stimulating HepG2.2.15 cells with palmitic acid and oleic acid to study the relationship between HBV-DNA load and lipid metabolism.The results of the in vitro experiments suggested that fatty acid treatment increased lipid droplet deposition and decreased the expression of cell supernatant HBsAg,HBeAg,and HBV DNA load.Western blot and polymerase chain reaction analysis showed that fatty acid stimulation significantly induced Plin2 protein expression and inhibited the expression of hepatocyte autophagy proteins.Inhibition of Plin2 protein expression under fatty acid stimulation reversed the reduction in HBsAg and HBeAg expression and HBV DNA load induced by fatty acid stimulation and the inhibition of cellular autophagy.Knocking down Plin2 and blocking autophagy with 3-methyladenine(3-MA)inhibited HBV DNA replication.CONCLUSION In conclusion,lipid metabolism is a significant factor affecting HBV load in patients with HBV infection.The in vitro experiments established that fatty acid stimulation inhibits HBV replication via the Plin2-autophagy pathway.展开更多
基金supported by grants from the Youth Innovative Talents Program and Characteristic Innovative Talents Program of Regular Institutions of Higher Education of Guangdong Province(2018KQNCX212,2020KTSCX101)the Scientific Research Project of Traditional Chinese Medicine Bureau of Guangdong Province(20222122)+3 种基金the Excellent Youth Science Research Program of the Education Department of Hunan Province(14B183)Bureau of Education of Guangzhou Municipality(202032801)the Plan on Enhancing Scientific Research in Guangzhou Medical Universitythe Open Research Funds(2021)and the Funds of Selected Project(2022)from GMU-GIBH Joint School of Life Sciences,Guangzhou Medical University。
文摘目的oxLDL可上调Plin2的表达,进而促进泡沫细胞的形成,LOX1是oxLDL的受体。本文探讨Plin2与LOX1在动脉粥样硬化发生发展过程中的关系。方法从GEO数据库中下载GSE43292,分析Plin2、LOX1的表达及Plin2、LOX1与NF-κB信号通路的相关性。采用oxLDL处理的RAW264.7细胞作为动脉粥样硬化的细胞模型进行研究,蛋白质免疫印迹法检测细胞中Plin2、LOX1和p-p65的表达,荧光中性脂质染料BODIPY 493/503染色法检测细胞内脂滴。结果通过分析GSE43292数据发现,Plin2、LOX1在颈动脉粥样硬化斑块中的表达显著高于颈动脉邻近组织。oxLDL处理RAW264.7细胞24 h后,Plin2与LOX1的表达、细胞内脂滴明显增加。过表达Plin2的细胞中LOX1表达升高;当用oxLDL孵育过表达Plin2的细胞后,LOX1的水平升高更为显著;但在没有oxLDL处理的情况下,敲减Plin2对细胞内LOX1的表达没有影响。基因集富集分析(gene set enrichment analysis,GSEA)结果显示,在动脉粥样硬化中,Plin2和LOX1的表达与NF-κB的活化呈正相关。此外,尽管采用oxLDL处理细胞,NF-κB抑制剂JSH-23预处理仍可显著降低Plin2与LOX1的表达、细胞内的脂质积聚,过表达Plin2后,JSH-23亦能显著抑制oxLDL孵育的细胞中Plin2和LOX1的表达。结论Plin2可通过上调LOX1的表达促进细胞内脂质积聚,参与动脉粥样硬化,这一过程至少部分是通过激活NF-κB通路实现的。
文摘BACKGROUND Chronic hepatitis B virus(HBV)infection is often associated with increased lipid deposition in hepatocytes.However,when combined with non-alcoholic fatty liver disease or hyperlipidemia,it tends to have a lower HBV deoxyribonucleic acid(DNA)load.The relationship between lipid metabolism and HBV DNA replication and its underlying mechanisms are not well understood.AIM To investigate the relationship between lipid metabolism and HBV DNA replication and its underlying mechanisms.METHODS 1603 HBsAg-seropositive patients were included in the study.We first explored the relationship between patients'lipid levels,hepatic steatosis,and HBV DNA load.Also,we constructed an HBV infection combined with a hepatic steatosis cell model in vitro by fatty acid stimulation of HepG2.2.15 cells to validate the effect of lipid metabolism on HBV DNA replication in vitro.By knocking down and overexpressing Plin2,we observed whether Plin2 regulates autophagy and HBV replication.By inhibiting both Plin2 and cellular autophagy under high lipid stimulation,we examined whether the Plin2-autophagy pathway regulates HBV replication.RESULTS The results revealed that serum triglyceride levels,high-density lipoprotein levels,and hepatic steatosis ratio were significantly lower in the HBV-DNA high load group.Logistic regression analysis indicated that hepatic steatosis and serum triglyceride levels were negatively correlated with HBV-DNA load.Stratified analysis by HBeAg showed significant negative correlations between HBV-DNA load and hepatic steatosis ratio in both HBeAgpositive and HBeAg-negative groups.An in vitro cell model was developed by stimulating HepG2.2.15 cells with palmitic acid and oleic acid to study the relationship between HBV-DNA load and lipid metabolism.The results of the in vitro experiments suggested that fatty acid treatment increased lipid droplet deposition and decreased the expression of cell supernatant HBsAg,HBeAg,and HBV DNA load.Western blot and polymerase chain reaction analysis showed that fatty acid stimulation significantly induced Plin2 protein expression and inhibited the expression of hepatocyte autophagy proteins.Inhibition of Plin2 protein expression under fatty acid stimulation reversed the reduction in HBsAg and HBeAg expression and HBV DNA load induced by fatty acid stimulation and the inhibition of cellular autophagy.Knocking down Plin2 and blocking autophagy with 3-methyladenine(3-MA)inhibited HBV DNA replication.CONCLUSION In conclusion,lipid metabolism is a significant factor affecting HBV load in patients with HBV infection.The in vitro experiments established that fatty acid stimulation inhibits HBV replication via the Plin2-autophagy pathway.
