摘要
目的探究microRNA-29b(miR-29b)在肝纤维化过程中的作用,以及对肝星状细胞增殖和凋亡的影响。方法将大鼠随机分为对照组和模型组,每组10只。模型组大鼠经腹部注射60%3 ml/kg CCl4复制肝纤维化模型;对照组注射等量生理盐水。分别观察两组大鼠肝纤维化情况,检测肝组织中miR-29b、TGF-β1、SAMD3基因及TGF-β1、SAMD3蛋白的表达。体外培养肝星状细胞HSC-T6、LX-2,转染miR-29b siRNA,检测miR-29b对HSC-T6、LX-2细胞增殖、凋亡的影响,以及对TGF-β1、SAMD3表达的影响。结果与对照组相比,模型组心肌纤维比例增加,随着时间延长,第6和12周心肌纤维比例逐渐升高(P <0.05)。与对照组相比,模型组miR-29b基因相对表达量降低,随着时间延长,模型组miR-29b基因相对表达量逐渐降低(P<0.05)。与对照组相比,模型组TGF-β1、SAMD3基因和蛋白相对表达量升高;随着时间延长,TGF-β1、SAMD3基因和蛋白相对表达量逐渐升高(P <0.05)。在HSC-T6、LX-2细胞中,miR-29b转染组G1期细胞比例高于空白对照组、阴性转染组,S期细胞比例低于空白对照组、阴性转染组(P<0.05)。miR-29b转染组HSC-T6、LX-2细胞凋亡率高于空白对照组、阴性转染组(P<0.05)。与空白对照组、阴性转染组相比,HSC-T6、LX-2细胞中TGF-β1、SAMD3基因和蛋白相对表达量升高,miR-29b基因相对表达量降低(P<0.05)。结论 miR-29b在肝纤维化过程中表达降低,miR-29b可抑制肝星状细胞增殖,诱导其凋亡。
Objective To explore the effect of microRNA-29b(miR-29b)in the process of hepatic fibrosis and its influence on the proliferation and apoptosis of hepatic stellate cells.Methods The experimental rats were divided into two groups,including a control group and a model group,with 10 cases in each group.The rats in the model group were injected with 60%3 ml/kg CCl4 by abdominal approach,and those in the the control group were injected with the same amount of normal saline.Liver fibrosis was observed in the control group and the model group.The expressions of miR-29b,TGFB1 and SAMD3 genes,the expressions of TGF-β1 and SAMD3 proteins were detected in the liver tissue.Hepatic stellate cells HSC-T6 and LX-2 were cultured in vitro and transfected with miR-29b siRNA,and the effect of miR-29b on proliferation and apoptosis of HSC-T6 and LX-2 cells,and on the expressions of TGF-β1 and SAMD3 were detected.Results Compared with the control group,the proportion of myocardial fibers increased in the model group,and with the prolongation of time that increased gradually in the 6th and 12th weeks(P<0.05).Compared with the control group,the relative expression of miR-29b gene in the model group decreased,and decreased gradually with the prolongation of time(P<0.05).Compared with the control group,the relative expressions of TGFB1 and SAMD3 genes and proteins increased in the model group,and gradually increased with the prolongation of time(P<0.05).The G1-phase cell percentage of HSC-T6 and LX-2 cells in the miR-29b transfection group was higher than that in the control group and the negative transfection group,and S-phase cell ratio was lower than that in the control group and the negative transfection group(P<0.05).The apoptosis rate of HSC-T6 and LX-2 cells in the miR-29b transfection group was higher than that in the control group and the negative transfection group(P<0.05).Compared with the control group and the negative transfection group,the relative expression of TGFB1 and SAMD3 mRNAs and proteins in HSC-T6 and LX-2 cells increased,and the relative expression of miR-29b mRNA decreased in the miR-29b transfection group(P<0.05).Conclusions The expression of miR-29b decreases in the process of hepatic fibrosis,and miR-29b can inhibit proliferation and induce apoptosis of hepatic stellate cells.
作者
王艳景
谷征
李圆圆
Yan-jing Wang;Zheng Gu;Yuan-yuan Li(Department of Infectious Diseases,Huaihe Hospital of Henan University,Kaifeng,Henan 475000,China)
出处
《中国现代医学杂志》
CAS
2018年第32期17-24,共8页
China Journal of Modern Medicine
