miR-29在乙型肝炎肝硬化患者中的表达及对肝星状细胞活化和上皮间充质转化的影响

Expression of miR-29 in patients with hepatitis B cirrhosis and its effect on activation of hepatic stellate cells and epithelial mesenchymal transition

目的 探讨miR-29在乙型肝炎肝硬化患者中的表达及对肝星状细胞(HSC)活化和上皮间充质转化(EMT)的影响。方法 选择2020年6月至2022年6月应城市人民医院收治的104例乙型肝炎患者作为研究对象,其中单纯乙型肝炎患者52例(设为慢性乙肝组),乙型肝炎合并肝硬化患者52例(设为乙肝肝硬化组)。采用实时荧光定量PCR法测定血清miR-29a、mi R-29b和miR-29c的表达水平,并分析miR-29与乙型肝炎肝硬化患者Child-Pugh分级和临床分期的关系。用miR-29a/b/c重组慢病毒感染人HSC系LX-2,制备过表达miR-29细胞模型,并分析过表达miR-29对LX-2细胞增殖、活化和EMT的影响。结果 与慢性乙肝组相比,乙肝肝硬化组血清miR-29a、miR-29b和miR-29c的表达水平均显著降低(P均<0.01)。乙型肝炎肝硬化患者血清miR-29a、miR-29b、mi R-29c的表达水平均随着ChildPugh分级增高及临床分期加重而逐渐降低,组间差异均有统计学意义(P均<0.01)。Pearson相关性分析结果显示,乙肝肝硬化组血清miR-29a、miR-29b、miR-29c的表达水平与Child-Pugh分级和临床分期均呈显著负相关(P均<0.01)。与空白对照组相比,重组慢病毒组的细胞增殖率显著降低,且α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(COLⅠ)、Ⅲ型胶原(COLⅢ)、转化生长因子-β1(TGF-β1)、波形蛋白(Vimentin)、N-钙黏蛋白(N-cadherin)的蛋白及其m RNA的表达水平均显著降低,E-钙黏蛋白(E-cadherin)的蛋白及其mRNA的表达水平均显著升高,组间差异均有统计学意义(P均<0.01)。结论 乙型肝炎肝硬化患者血清miR-29的表达水平较低,且其与Child-Pugh分级和临床分期相关;过表达mi R-29可能通过抑制HSC增殖、活化及EMT而发挥抗肝纤维化作用。

Objective This paper is to investigate the expression of miR-29 in patients with hepatitis B cirrhosis and its effect on the activation of hepatic stellate cells(HSC)and epithelial mesenchymal transition(EMT).Methods A total of 104 patients with hepatitis B admitted to Yingcheng People's Hospital from June 2020 to June 2022 were selected,including 52 patients with simple hepatitis B(set as the chronic hepatitis B group)and 52 patients with hepatitis B complicated with cirrhosis(set as the hepatitis B cirrhosis group).The expression of serum miR-29a,miR-29b,and miR-29c was detected by real-time fluorescence quantitative PCR,to analyze the relationship between miR-29 and Child-Pugh grading and clinical staging in patients with hepatitis B cirrhosis.Using miR-29a/b/c recombinant lentivirus to infect human HSC line LX-2,an overexpression miR-29 cell model was prepared,and the effects of the overexpression miR-29 on LX-2 cell proliferation,activation,and EMT were analyzed.Results Compared with the chronic hepatitis B group,the expression of serum miR-29a,miR-29b,and miR-29c in the hepatitis B cirrhosis group is lower,with statistically significant differences(P<0.01).The expression of serum miR-29a,miR-29b,and miR-29c in patients with hepatitis B cirrhosis is gradually decreased with the increase of Child-Pugh grade and clinical stage,with statistically significant differences(P<0.01).Pearson correlation analysis shows that the expression of serum miR-29a,miR-29b,and miR-29c in patients with hepatitis B cirrhosis is significantly negatively correlated with Child-Pugh grade and clinical stage(P<0.01).Compared with the blank control group,the cell proliferation rate of the recombinant lentivirus group is reduced,and the expression ofα-smooth muscle actin(α-SMA),typeⅠcollagen(COLⅠ),typeⅢcollagen(COLⅢ),transforming growth factor-β1(TGF-β1),Vimentin,N-cadherin protein,and mRNA is decreased,while the expression of E-cadherin protein and mRNA is increased,with statistically significant differences(P<0.01).Conclusions The expression of serum miR-29 in patients with hepatitis B cirrhosis is low,and is related to Child-Pugh grade and clinical stage.Overexpression of miR-29 may play an anti-hepatic fibrosis role by inhibiting the proliferation,activation of HSC and EMT.