摘要
目的:研究miR-377靶向调控早期生长反应因子(early growth response l,Egr1)在肝纤维化病理机制中的作用。方法:RT-qPCR检测肝纤维化患者血清中miR-377的表达;用miR靶基因数据库进行筛选,选择Egr1为miR-377的潜在靶基因;RT-qPCR和Western印迹法分别检测miR-377 mimics对Egr1的mRNA及蛋白表达的影响;CCK-8检测Ang Ⅱ诱导后HSC细胞和转染miR-377的HSC细胞增殖程度;RT-qPCR检测经AngⅡ和AngⅡ+miR-377刺激后与α-平滑肌动蛋白(α-smooth muscle actin,α-SM A),FSP1和Col lagenI的表达量。结果:MiR-377可能与肝纤维化的发生及发展有关;miR-377可以通过结合Egr1的3'-UTR直接抑制Egr1的mRNA翻译及其蛋白表达;miR-377可抑制肝细胞经AngⅡ诱导后的增殖活性;miR-377能抑制经AngⅡ刺激后α-SMA,FSP1和CollagenI的表达,在肝纤维化的进程中发挥抑制作用。结论:MiR-377通过抑制Egr1的表达抑制肝细胞的增殖和病理进程。
Objective:To study the role of miR-377 targeted regulation of EGR1 in the pathological mechanism of liver fibrosis.Methods:The expression of miR-377 in serum of patients with liver fibrosis were detected by RT-qPCR.Egr1 was selected as the potential target gene of miR-377 by screening with miR target gene database.The effect of miR-377mimics on the mRNA and protein expression of Egr1 was detected by RT-qPCR and Western blot,respectively.CCK-8 was used to detect the proliferation of HSC cells and HSC cells transfected with miR-377 which were induced by AngII.RT-qPCR was used to detect the expression ofα-smooth muscle actin(α-SMA),FSP1 and Collagen I after AngII and AngII+miR-377 stimulated.Results:MiR-377 may be related to the occurrence and development of liver fibrosis.miR-377 can directly inhibit the mRNA translation and protein expression of Egr1 by binding 3'-URT of Egr1.miR-377 could inhibit the proliferation of liver cells induced by AngII.miR-377 could inhibit the expression ofα-SMA,FSP1 and Collagen I after AngII stimulated and miR-377 played an inhibitory role in the process of liver fibrosis.Conclusion:MiR-377 inhibits the proliferation and pathological process of liver cells by inhibiting the expression of Egr1.
作者
李贻弘
张宏
侯波
熊首先
LI Yihong;ZHANG Hong;HOU Bo;XIONG Shouxian(Department of Gastroenterology,Jianghan Oilfield General Hospital,Qianjiang Hubei 433124,China)
出处
《临床与病理杂志》
2019年第3期479-485,共7页
Journal of Clinical and Pathological Research
