【摘要】 该文探讨肝实质细胞中整合素α5(ITGA5)对醛脱氢酶4A1(ALDH4A1)表达的影响。利用DNA羟甲基化测序、RNA测序及免疫组织化学分析特异靶向敲减小鼠肝实质细胞Itga5对ALDH4A1基因启动子羟甲基化水平、m RNA及蛋白水平的影响。利用免疫组织化学染色技术分析胆汁性肝硬化患者和正常供者肝组织ITGA5及ALDH4A1的蛋白水平。利用siRNA敲减人肝实质细胞中ITGA5,利用实时定量RT-PCR及免疫细胞化学染色技术,分析敲减ITGA5对人肝实质细胞中ALDH4A1 mRNA及蛋白水平的影响。结果显示,特异敲减小鼠肝实质细胞中Itga5,肝脏组织中ALDH4A1基因启动子的羟甲基化水平及蛋白水平明显高于对照组,差异有统计学意义(P<0.05)。同样,敲低人肝实质细胞中ITGA5的表达水平, ALDH4A1的mRNA水平和蛋白水平也明显高于对照组(P<0.05)。胆汁性肝硬化患者的肝脏组织中ITGA5蛋白水平低于正常供者,而ALDH4A1蛋白水平高于正常供者,差异有统计学意义(P<0.05)。研究表明,肝实质细胞中整合素α5可能通过调节基因启动子羟甲基化水平调节ALDH4A1的表达。更多还原

【Abstract】 This study investigates the effect of ITGA5(integrin α5) in hepatocytes on the expression of ALDH4A1(aldehyde dehydrogenase 4A1). DNA hydroxymethylation sequencing, RNA sequencing, and immunohistochemical analysis were employed to examine the impact of specifically targeted knockdown of Itga5 in mouse hepatocytes on the hydroxymethylation level of the ALDH4A1 gene promoter, as well as its mRNA and protein levels. Immunohistochemical staining was used to analyze the protein levels of ITGA5 and ALDH4A1 in liver tissues from patients with biliary cirrhosis and normal donors. siRNA was utilized to knockdown ITGA5 in human hepatocytes, and real-time quantitative RT-PCR along with immunocytochemical staining were performed to assess the effect of ITGA5 knockdown on ALDH4A1 mRNA and protein levels in human hepatocytes. The results showed that specific knockdown of Itga5 in mouse hepatocytes significantly increased the hydroxymethylation level of the ALDH4A1 gene promoter and its protein level in liver tissues compared to the control group, with a statistically significant difference(P<0.05). Similarly, knockdown of ITGA5 in human hepatocytes also led to significantly higher m RNA and protein levels of ALDH4A1 compared to the control group(P<0.05). In liver tissues from biliary cirrhosis patients, ITGA5 protein levels were lower than those in normal donors, whereas ALDH4A1 protein levels were higher, with statistically significant differences(P<0.05). The study suggests that integrin α5 in hepatocytes may regulate the expression of ALDH4A1 by modulating the hydroxymethylation level of the gene promoter.更多还原