Changes Of LncRNA Expression Profile And Function In Hepatic Stellate Cells Induced By Hypoxia

Backgrounds&Aims:Liver fibrosis can be caused by a variety of chronic liver damage and can subsequently progress to cirrhosis and hepatocellular carcinoma.During the development of liver fibrosis,hepatic stellate cell?HSC?activation plays an important role.Hypoxia plays an important role in HSC activation and subsequent liver fibrosis.In addition,the role of long non-coding RNA?lncRNA?in liver fibrosis has been gradually paid attention to,but there are still few studies on the changes of lncRNA expression profile and its function under hypoxia.Therefore,in this study,cobalt dichloride?CoCl₂?was used to induce cell hypoxia.On the one hand,the expression of factors related to HSC activation was detected,on the other hand,the whole genome sequencing analysis was used to analyze the changes of lncRNA expression profile and its function in HSC after hypoxia treatment,in order to provide a new idea for the diagnosis and treatment of liver fibrosis.Methods:HSC was cultured in vitro,rat HSC was treated with CoCl₂ of different concentrations,and the effect of CoCl₂ on the activity of rat HSC was detected by cck-8 method.RT-qPCR was used to determine hypoxia induction factor-1??HIF-1??,?-smooth muscle actin??-SMA?,type I collagen?collagen I?,transforming growth factor-?1?TGF?-1?and Smad3 mRNA expression.At the same time,HSC were treated with 400?mol/L CoCl₂ and whole genome sequencing was used to analyze the changes of lncRNA expression profiles in HSC after CoCl₂treatment.The differentially expressed lncRNAs were analyzed for gene ontology?GO?and kyoto gene and genome encyclopedia?KEGG?analysis.Results:Different concentrations of CoCl₂ have different inhibitory effects on the activity of rat HSC,and the activity of HSC decrease with the increase of the concentration of CoCl₂ and the extension of the action time.After hypoxia treatment,the expression of HIF-1?,?-SMA,collagen I,TGF-?1,and Smad3 in HSC was significantly higher than that in the untreated group.The HSC whole genome sequencing results showed that,compared with the untreated group,137 lncRNAs were significantly up-regulated in HSC,while 151 lncRNAs were significantly down-regulated after hypoxia treatment.In addition,bioinformatics analysis showed that the differentially expressed lncRNA was mainly concentrated in the processes of cell hypoxia,cell differentiation,proliferation,and deposition of extracellular matrix?ECM?.Conclusion:This study found that hypoxia can induce HSC activation and up-regulate the expression of liver fibrosis-related genes.In addition,hypoxia can cause differential expression of lncRNA in HSC,revealing the key role of lncRNA in HSC activation,suggesting that lncRNA may become new diagnostic markers and targets for future liver fibrosis.