Effects of fasudil on inhibiting vascular smooth muscle cell proliferation and its molecular mechanism / 中华老年医学杂志

ABSTRACT

Objective To explore the effects of fasudil on inhibiting vascular smooth muscle cells (VSMCs) proliferation in vitro,increasing cell apoptosis,and inhibiting the Ras-MEK 1/2-ERK 1/2 pathway.Methods Healthy male SD rats (80～100 g) were selected.VSMCs were separated by the thoracoabdominal aortic vascular membrane dissection.Cultured VSMCs were randomly divided into 5 groupsserum-free group; serum group; serum + 1 μmol/L fasudil intervention group; serum + 10 μmol/L fasudil intervention group; serum + 100 μmol/L fasudil intervention group.The proliferation and migration of VSMCs were detected by MTT method and wound healing assay.Cell cycle and apoptosis of VSMCs were examined by flow cytometric analysis.The mRNA expressions of pro-apoptotic protein (Bax) and anti-apoptotic protein(Bcl-2) were determined by RT-PCR method,and the ratio of Bax/Bcl 2 was calculated.Western blot were performed to detect the protein expressions of Ras,MEK1/2,ERK1/2 and Akt in VSMCs.Results Fasudil inhibited rat VSMCs proliferation and migration,and blocked FBS-induced progression from the G0/G1 phase to S phase in a dose-dependent manner.Fasudil inhibited the early and late apoptosis in VSMCs,increased Bax mRNA expression and inhibited Bcl 2 mRNA expression.Fasudil significantly inhibited the protein expressions of FBS-stimulated intracellular Ras,phosphorylated MEK1/2,ERK1/2 in a dosedependent manner,but did not affect the protein expression of phosphorylated Akt.Conclusions Fasudil can attenuate VMSCs proliferation by blocking Ras-MEK1/2-ERK1/2 pathway and increasing cell apoptosis.