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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 investigate the effects of angiotensin Ⅱ (Ⅱ) on the transdifferentiation of peritoneal mesothelial cells (RPMCs) and the second generation of peritoneal mesothelial cells pretreated with astragalus injection (AG). Methods: The primary cultured RPMCs of SD rats were cultured in vitro for 2 generations. After being kept for 24 hours, they were randomly divided into 4 groups: normal control group, AngⅡ (1 × 10-7mol / L) group, AngⅡ + AG AngⅡ + AG (2g / mL) group. In Ang Ⅱ (1 × 10-7mol / L) group, RPMCs were stimulated with Ang (1 × 10-7mol / L) in vitro. After being pretreated with AG (1 g / mL) and AG (2 g / mL) for 1 h in AngⅡ + AG and AngⅡ + AG groups, Ang Ⅱ (1 × 10-7 mol / L ) Stimulate RPMCs in vitro. The expression of α-SMA and E-Cadherin mRNA was detected by real-time quantitative RT-PCR. The expression of α-SMA and E-Cadherin mRNA was detected by Western blotting. Gene expression in 8h, protein expression in 24h, respectively, the above groups of genes and protein expression levels were observed. Results: Ang Ⅱ induced up-regulated mRNA and protein expression of translocation marker-α-SMA in peritoneal mesothelial cells, both P <0.05. AG pretreatment inhibited Ang-induced α-SMA expression and decreased α-SMA mRNA and protein levels, both P <0.05. No inhibition of α-SMA mRNA and protein level by AG was observed The concentration of a relationship. AngⅡinduced the down-regulation of E-Cadherin mRNA and protein, AG significantly reversed the downregulation of this expression (P <0.05, P <0.01), and did not observe the effect of AG on E-Cadherin mRNA and protein expression The effect is related to the concentration of AG. Conclusion: AG can inhibit the transdifferentiation of peritoneal mesothelial cells to mesenchymal cells, which provides a basis for its prevention and treatment of peritoneal fibrosis.