异丙酚抑制血管紧张素Ⅱ诱导的心肌成纤维细胞增殖的机制研究

Study on the mechanism of propofol inhibiting the proliferation of cardiac fibroblasts induced by angiotensionⅡ

目的:探讨异丙酚(Propofol)抑制血管紧张素Ⅱ(AngⅡ)诱导的大鼠心肌成纤维细胞(CFb)增殖的机制。方法:用AngⅡ建立诱导新生大鼠CFb纤维化模型。将心肌细胞分为对照组、AngⅡ组和异丙酚组。用流式细胞仪测定各组细胞的周期分布;用ELISA法测定各组细胞中Ⅰ、Ⅲ型胶原含量;用Western Blot法测定各组细胞中转化生长因子β1(TGF-β1)的蛋白表达量;用免疫细胞化学化法测定各组细胞中细胞外信号调节蛋白激酶(ERK1/2)和经典型蛋白激酶Cα亚型(cPKCα)的表达。结果:100μmol/L的异丙酚对AngⅡ诱导的大鼠CFb的增殖抑制效果最显著(P<0.01)。与对照组相比,AngⅡ组细胞G 0/G 1期细胞百分率降低(P<0.01),S期和G 2/M期细胞百分率升高(P<0.01);异丙酚组细胞的G 0/G 1期细胞百分率高于AngⅡ组(P<0.01),S期和G 2/M期细胞百分率低于AngⅡ组(P<0.01)。与对照组相比,AngⅡ组大鼠的CFbⅠ、Ⅲ型胶原含量增加(P<0.01);异丙酚组大鼠的CFbⅠ、Ⅲ型胶原含量低于AngⅡ组(P<0.01)。与对照组相比,AngⅡ组大鼠CFb中TGF-β1的蛋表达升高(P<0.01),异丙酚组大鼠CFb中TGF-β1的表达低于AngⅡ组(P<0.01)。与对照组相比,AngⅡ组大鼠CFb中ERK1/2和cPKCα的表达增加(P<0.01);异丙酚组大鼠CFb中ERK1/2和cPKCα的表达低于AngⅡ组。结论:异丙酚能够抑制AngⅡ诱导的大鼠CFb的增殖和Ⅰ、Ⅲ型胶原含量的增加,其机制与降低细胞中TGF-β1的蛋白表达和抑制ERK1/2、cPKCα通路有关。

Objective:To investigate the mechanism of propofol inhibiting angiotensin II(Ang II)-induced proliferation of rat cardiac fibroblasts(CFb).Methods:Ang II was used to establish a model of CFb fibrosis in neonatal rats.Cardiomyocytes were divided into control group,Ang II group and propofol group.The cell cycle distribution was determined by flow cytometry and the content of collagen type I and collagen type III was determined by ELISA.The protein expression of transforming growth factor beta 1(TGF-beta 1)in the cells of each group was determined by Western Blot.The expression of extracellular signal-regulated protein kinase 1/2(ERK1/2)and typical protein kinase C alpha subtype(cPKCalpha)were determined by immunocytochemistry.Results:Propofol at 100 mol/L had the most significant inhibitory effect on Ang II induced proliferation of CFb in rats(P<0.01).Compared with the control group,the percentage of G 0/G 1 phase cells in Ang II group decreased(P<0.01),and the percentage of S phase and G 2/M phase cells increased(P<0.01).The percentage of G 0/G 1 phase cells in propofol group was higher than that in Ang II group(P<0.01),and the percentage of S phase cells and G 2/M phase cells in propofol group was lower than that in Ang II group(P<0.01).Compared with the control group,the concentration of CFb type I and III collagen increased in Ang II Group(P<0.01).The concentrations of CFb type I and III collagen in the propofol group were lower than those in Ang II group(P<0.01).Compared with the control group,the expression of TGF-β1 in CFb of Ang II group increased(P<0.01),and that of TGF-β1 in CFb of propofol group was lower than that of Ang II group(P<0.01).Compared with the control group,the expression of ERK1/2 and cPKCαin CFb of Ang II group increased(P<0.01).The expression of ERK1/2 and cPKC alpha in CFb of propofol group was lower than that of Ang II group.Conclusion:Propofol could inhibit the proliferation of CFb induced by Ang II and increase the concentration of type I and III collagen.The mechanism is related to reducing the expression of TGF-beta 1 and inhibiting the ERK1/2 and cPKC alpha pathway.