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目的:观察肝脏特异性SIRT1敲除(SIRT1-LKO)小鼠在四氯化碳(CCL4)诱导下的肝纤维化情况,系统地探讨SIRT1及转录差异基因在肝纤维化中的作用和机制。方法:利用Cre-Lox P重组酶系统构建SIRT1-LKO小鼠模型,经腹腔注射CCL4橄榄油溶液来诱导小鼠肝纤维化,通过血清生化检测肝功能,使用天狼星红染色观察肝脏胶原蛋白沉积,检测α-平滑肌肌动蛋白(α-SMA)的表达来观察肝星状细胞(HSCs)的活化,并进一步利用基因芯片技术和生物信息学分析来筛选转录差异基因。结果:在CCL4诱导下,SIRT1-LKO小鼠比同窝野生(WT)小鼠的肝损伤更加严重,肝纤维化也更为显著(P<0.05);通过对转录差异基因进行GO生物过程和KEGG通路分析,发现了一组可能与SIRT1和肝纤维化都存在相关的关键基因(TNC、TPM1、E2F1、DEFB1、LRTM1)。结论:SIRT1缺失会增加CCL4诱导的小鼠肝损伤,加重肝纤维化;SIRT1可能与上述基因协同参与了肝纤维化的发生发展。
OBJECTIVE: To observe the hepatic fibrosis induced by carbon tetrachloride (CCL4) in liver-specific SIRT1 knockout (SIRT1-LKO) mice and to explore the role and mechanism of SIRT1 and transcriptional difference genes in hepatic fibrosis. Methods: The SIRT1-LKO mouse model was constructed by using Cre-Lox P recombinase system, and the hepatic fibrosis was induced by intraperitoneal injection of CCL4 olive oil solution. The hepatic function was detected by serum biochemistry and collagen deposition was observed by Sirius red staining. The expression of α-smooth muscle actin (α-SMA) was detected to observe the activation of hepatic stellate cells (HSCs), and the genes of transcriptional differences were further screened by gene chip technology and bioinformatics analysis. Results: Liver injury was more severe and liver fibrosis was more severe in SIRT1-LKO mice than in wild type (WT) mice under CCL4 induction (P <0.05). Through the GO biological process and KEGG pathway analysis revealed a group of key genes (TNC, TPM1, E2F1, DEFB1, LRTM1) that may be involved in both SIRT1 and hepatic fibrosis. CONCLUSIONS: SIRT1 deficiency increases liver injury induced by CCL4 in mice and aggravates liver fibrosis. SIRT1 may be involved in the development of hepatic fibrosis with the above genes.