阿托伐他汀抑制内皮细胞Rh0/Rho激酶信号通路改善细胞骨架

Atorvastatin inhibits Rho/Rho kinase signal pathway and improves endothelial cytoskeleton

目的探讨阿托伐他汀对血管紧张素Ⅱ(AngⅡ)诱导的人脐静脉内皮细胞(HUVEC)Rho/Rho激酶信号通路中Rho激酶和磷酸化肌球蛋白轻链(p-MLC)表达的影响。方法体外培养HUVEC,分4组:对照组;AngⅡ组;阻断剂组(Rho激酶阻断剂Y-27632);阿托伐他汀组。硝酸还原酶法检测NO含量;免疫细胞化学法观察p-MLC蛋白定位表达,Western blot法测定Rho激酶和p-MLC蛋白定量表达。结果与对照组比较,AngⅡ组NO含量显著减少(P<0.01);阻断剂组和阿托伐他汀组NO含量较AngⅡ组显著升高(P<0.01)。AngⅡ组细胞质内出现大量棕色颗粒积聚、浓染;阻断剂组和阿托伐他汀组细胞质中仅有少量棕色淡染颗。与AngⅡ组比较,阻断剂组和阿托伐他汀组Rho激酶及p-MLC蛋白表达显著降低(P<0.01),但仍高于对照组(P<0.01);2组间蛋白表达仍有明显差异(P<0.01)。结论阿托伐他汀对AngⅡ诱导的HUVEC保护作用,是通过抑制Rho/Rho激酶信号通路中Rho激酶及其下游的p-MLC蛋白表达,减弱AngⅡ对内皮细胞骨架的损伤。

Ojective To study the effect of atorvastatin on expression of Rho kinase and phospho- rylated myosin light chain（p-MLC） in angiotensin Ⅱ （Ang Ⅱ ）-induced Rho/Rho kinase signal pathway of human umbilical vein endothelial cells （HUVEC）. Methods HUVEC, cultured in vitro,were divided into normal control group, Ang Ⅱ group, Rho kinase blocker Y-27632 group, and atorvastatin group. Nitric oxide（NO） level in HUVEC was measured by nitrate reduction test. Expression of p-MLC protein and Rho kinase in HUVEC was detected by immunocytochem- istry and Western blot, respectively. Results The NO level was significantly lower in Ang Ⅱ group than in control group and significantly higher in Rho kinase blocker Y-27632 group than in atorvastatin group（P〈0.01）. A large number of brown and hyperchromatic granules were accu- mulated in cytoplasm of Ang Ⅱ group while a small number of brown and hypochromatic granules were accumulated in cytoplasm of Rho kinase blocker Y-27632 group and atorvastatin group. Al- though the expression level of Rho kinase and p-MLC protein decreased significantly in Rho ki- nase blocker Y-27632 group and Ang Ⅱ group, it was still higher than that in control group（P〈 0. 01）. Conclusion Atorvastatin can protect Ang H-induced HUVEC by controlling the expression of Rho kinase and its downstream p-MLC protein in Rho/Rho kinase signaling pathway and weakening the Ang Ⅱ-induced endothelial cytoskeleton injury.