黄芪注射液对AngⅡ诱导的大鼠腹膜间皮细胞转分化的干预作用

Effect of Astragalus Injection on epithelial-to-mesenchymal transition of rat peritoneal mesothelial cells induced by Angiotensin Ⅱ

目的:探讨AngⅡ诱导腹膜间皮细胞(RPMCs)转分化,及2代腹膜间皮细胞经黄芪注射液(AG)预处理后的干预作用。方法:体外培养SD大鼠原代RPMCs至2代,静止24h后,随机分为4组:正常对照组,AngⅡ(1×10-7mol/L)组,AngⅡ+AG(1g/mL)组和AngⅡ+AG(2g/mL)组。AngⅡ(1×10-7mol/L)组给予Ang(1×10-7mol/L)体外刺激RPMCs。AngⅡ+AG(1g/mL)组和AngⅡ+AG(2g/mL)组分别用AG(1g/mL)、AG(2g/mL)预处理1h后,再行AngⅡ(1×10-7mol/L)体外刺激RPMCs。实时定量RT-PCR法测定α-SMA、E-CadherinmRNA的表达;Western印迹法测定α-SMA、E-Cadherinm蛋白的表达。基因表达在8h,蛋白表达在24h,分别观察以上各组基因和蛋白的表达水平。结果:AngⅡ可诱导腹膜间皮细胞转分化标志因子α-SMAmRNA和蛋白水平表达上调,二者均P<0.05。经AG预处理后可抑制由AngⅡ诱导的α-SMA表达上调,使α-SMAmRNA和蛋白水平均降低,二者均P<0.05,未观察到AG对α-SMAmRNA和蛋白水平的抑制程度与AG的浓度呈有关系。AngⅡ可诱导E-CadherinmRNA和蛋白的表达下调,AG可以显著逆转这种表达的下调,差异有统计学意义(P<0.05,P<0.01),亦未观察到AG对E-CadherinmRNA和蛋白表达的影响与AG的浓度有关。结论:AG对腹膜间皮细胞向间充质细胞转分化过程有抑制作用,为其防治腹膜纤维化提供了依据。

Objective：To explore the transdifferentiation of rat peritoneal mesothelial cells（RPMCs）induced by AngiotensinⅡ,and the effect of Astragalus Injection（AG）on RPMCs transdifferentiation.Methods：Primary RPMCs were cultivated to the second generation in vitro.After synchronization for 24 hours,the cells were randomly assigned to the control group,the AngⅡ（10-7 mol/L）group,the AngⅡ＋AG（1g/mL）group,the AngⅡ＋AG（2g/mL）group.AngⅡ（10-7 mol/L）group： RPMCs was stimulated in vitro with AngⅡ（10-7 mol/L）.the AngⅡ＋AG（1g/mL）group：RPMCs was stimulated after pretreatment with AG（1g/mL）,and the AngⅡ＋AG（2g/mL）group：with AG（2g/mL）for1hour,and RPMCs was stimulated in vitro with AngⅡ （10-7mol/L）.The expression of mRNA of a-SMA and E-Cadherin were detected by real-time quantitative reverse transcriptionpolymerase chain reaction（RT-PCR）method,and the protein expression of a-SMA and E-Cadherin were detected by western blot method.Observe the changes of each group,gene level at 8h and protein level at 24h.Results：AngⅡstimulated the a-SMA mRNA and protein synthesis of RPMCs,and which were ameliorated by AG,P0.05,Not observed that the inhibited degree for a-SMA mRNA and protein were positively correlated with AG concentration.AngⅡdown-regulated E-cadherin mRNA expression and protein synthesis in RPMCs,which were ameliorated by AG,differences were significant,P0.05,not observed the expression for E-Cadherin mRNA and protein were positively correlated with AG concentration.Conclusion：AG inhibited the process of epithelial-to-mesenchymal transition to protect the peritoneum from fibrosis.