Atorvastatin inhibits vascular smooth muscle cell phenotype transformation in cerebral hemorrhage rats by down regulating KLF-5

Objective: To investigate whether atorvastatin (ATV) could inhibit vascular smooth muscle cell (VSMCs) phenotype transformation by down regulating KLF-5 in cerebral hemorrhage (CH) rats. Methods: Cerebral hemorrhage animal model was established successfully, VSMCs were isolated by adhesion method, followed by primary cultured and identified. After 3-5 generations, cells were used for the subsequent experiment. The expressions of mature VSMCs marker proteins, including SM-actin (SMA), SM-22 alpha, SM-MHC and OPN were detected by Western blotting. The proliferation and migration of VSMCs were detected by MTT assay and wound healing method, respectively. The morphology of VSMCs was observed by detecting the expression of SMA by using cell immune fluorescence. Then KLF-5 over-expression or blank control plasmid were transfected into VSMCs in-vitro, phenotypic transformation of VSMCs were detected with ATV treatment by using the above-mentioned experimental methods. Results: MTT and wound healing results showed that compared with control group, VSMCs proliferation and migration were significantly reduced in ATV groups. Western blotting results showed that the expression of SMA and SM-MHC in ATV groups was increased, and the expression of OPN and KLF-5 was decreased significantly compared with control group. Cell immune fluorescence results showed that the morphology of VSMCs became thinner and longer in ATV groups when compared with control group. However, after transfection of KLF-5 over-expression plasmid, the proliferation and migration of VSMCs were significantly inhibited, cell morphology and the phenotype-associated proteins concentration were changed. Conclusion: ATV inhibited VSMCs phenotype transformation in CH rats by down regulating KLF-5 expression, which providing a new idea for gene therapy in the development of CH.