转录组测序分析非酒精性脂肪肝的重要调控分子

Transcriptomic profiling and analysis of key regulatory molecules in non-alcoholic fatty liver disease

目的探讨非酒精性脂肪肝(non-alcoholic fatty liver disease,NAFLD)细胞与正常肝细胞中差异表达的基因。方法建立NAFLD细胞模型。采用棕榈酸诱导人正常肝细胞LO2细胞;油红O染色观察细胞脂肪蓄积情况;流式细胞仪检测棕榈酸对LO2细胞的凋亡影响。对正常LO2细胞和模型细胞组进行转录组测序,筛选差异表达基因,并对差异表达基因进行KEGG功能富集分析和GO功能富集分析。采用RT-PCR检测差异基因的表达。结果随着棕榈酸浓度的增加,细胞脂肪蓄积越明显,结合凋亡率筛选出最优棕榈酸诱导浓度用于后续研究。转录组结果显示,与对照组相比,模型组差异表达基因数目有780个,上调基因数目447个,下调基因数目333个,GO富集通路分析具有显著富集意义,KEGG富集通路主要涉及了20条信号通路,主要包括TNF、蛋白输出、NOD等信号通路。RT-PCR结果显示FGF21、AMBP、GOLGA7B和DDIT3在NAFLD中高表达,SPTLC3、PALM2和SPTSBB在NAFLD中低表达。结论NAFLD可引起相关差异基因表达,可能通过脂质代谢中的关键靶基因调控NAFLD发生发展。

Objective To explore the differentially expressed genes in Non-alcoholic fatty liver disease(NAFLD)compared to normal liver cells,aiming to elucidate genetic varations contributing to the pathophysiology of NAFLD.Methods Establishing the NAFLD cell model:Human normal liver cells(LO2)were treated with palmitic acid to induce NAFLD characteristics.Subsequently,Oil red O staining was employed to assess cellular fat accumulation,and flow cytometry was utilized to evaluate the apoptotic effects of palmitic acid on LO2 cells.Transcriptome sequencing was performed on both normal LO2 cells and cells in the induced model to identify differentially expressed genes.These genes underwent KEGG and GO functional enrichment analysis.Additionally,the expression levels of these differential genes were quantified using RT-PCR.Results Increasing concentrations of palmitic acid led to more pronounced cellular fat accumulation.Considering this alongside the apoptosis rate,an optimal concentration of palmitic acid was selected for further study.Transcriptomic analysis revealed that,in the model group,there were 780 differentially expressed genes compared to the control group,with 447 genes upregulated and 333 downregulated.GO enrichment analysis demonstrated significant pathway involvement,and KEGG pathway analysis indicated major involvement in 20 signaling pathways,including TNF,protein export,and NOD signaling pathways.RT-PCR results showed that FGF21,AMBP,GOLGA7B,and DDIT3 were highly expressed in the NAFLD model,whereas SPTLC3,PALM2,and SPTSBB showed low expression.Conclusion NAFLD is observed to induce the expression of specific differential genes,potentially regulating the progression of NAFLD through key target genes involved in lipid metabolism.