瑞舒伐他汀抑制血管平滑肌细胞表型转化的机制研究

摘要：目的  研究瑞舒伐他汀是否可以抑制大鼠主动脉血管平滑肌细胞（VSMCs）表型转化及其机制。方法  SD大鼠主动脉VSMCs原代细胞培养鉴定，取4～7代细胞用于实验。用不同浓度的瑞舒伐他汀干预VSMCs，Western blot检测各组标本中骨骼肌肌动蛋白（SM-actin）、平滑肌22α（SM-22α）、滑肌肌球蛋白重链（SM-MHC）、骨桥蛋白（OPN）的表达，噻唑蓝（MTT）法检测细胞增殖情况，划痕试验观察细胞迁移情况。通过免疫组织化学法（ICC）检测SM-actin的表达从而观察VSMC的形态。转染过表达KLF-4或空白对照质粒至离体培养的VSMCs中，用瑞舒伐他汀干预，并采用上述实验方法检测VSMCs表型转化情况。结果  MTT结果显示，与对照组比较，瑞舒伐他汀组VSMCs增殖能力降低，差异有统计学意义（P <0.05）。划痕试验结果显示，与对照组比较，他汀组VSMCs迁移减少，差异有统计学意义（P <0.05）。Western blot检测显示，与对照组比较，他汀组VSMCs中SM-22α、SM-MHC、OPN表达增多，差异有统计学意义（P <0.05）。ICC结果显示，与对照组VSMCs的细胞形态比较，他汀组VSMCs形态变细、变长。转染KLF-4过表达质粒后，与对照组比较，转染KLF-4过表达质粒可以增加VSMCs的增殖和迁移，差异有统计学意义（P <0.05），逆转他汀对VSMCs表型转化有抑制作用。结论  瑞舒伐他汀通过下调KLF-4，抑制大鼠主动脉VSMCs表型转化。

Rosuvastatin inhibits phonotype transformation of vascular smooth muscle cells by down-regulation of KLF-4

Abstract: Objective To verify whether Rosuvastatin could inhibit vascular smooth muscle cell (VSMC) phonotype switch by down-regulation of KLF-4. Methods The primary culture and identification of SD rat VSMCs were conducted, VSMCs in the 3rd to 5th passages were used for the following experiments. After treatment with different concentrations of Rosuvastatin, MTT was used to investigate the proliferative ability of VSMCs. Transwell chambers and wound healing were employed to test the migration ability of VSMCs. IHC was used to detect the expression of SM-actin and the morphological structure of VSMCs. Western blot was used to investigate the expressions of SM-actin, SM-MHC, SM-22α, osteopontin (OPN) and KLF-4 in VSMCs. After KLF-4 was transfected into VSMCs, the metheods were used again to test the phonotype changes of the VSMCs. Results Compared with the control group, the migration and proliferation ability of the VSMCs were decreased in the Rosuvastatin group. The expressions of SM-actin, SM-22α and SM-MHC in the Rosuvastatin group were significantly increased (P < 0.05), and the expression of OPN also increased (P < 0.05). IHC showed the VSMCs in the Rosuvastatin group became thinner and longer than those of the control group. After KLF-4 was transfected into VSMCs, the proliferation and migration ability of the VSMCs increased (P < 0.05). Overexpression of KLF-4 reversed the effect of Rosuvsatatin on VSMCs phonotype transformation. Conclusions Rosuvastatin inhibits VSMCs phonotype switch by down-regulation of KLF-4.