Effect Of Fluvastatin On The Proliferation And Expression Of Apoptosis Related-Gene On Vascular Smooth Muscle Cells Induced By Lysophosphatidylcholine

Objective:The aim of this study was to clarify the effects of fluvastatin on the proliferation and expression of apoptosis related-gene on human umbilical artery vascular smooth muscle cells induced by lysophosphatidylcholine(LPC)and its mechanisms by using different experimental methods.This study can provide a evidence to statins in clinical application.Methods:The HUASMC from Embryo Artery vein was primary cultured and identified withα-actin.The HUASMC were divided into 4 groups:Control group(no stimulation),LPC(5 mg·L^-1)group,fluvastatin group:HUASMC were incubated with fluvastatin of different concentration(10^-5to 10^-7mol·L^-1)for 1 hour before stimulated with lysophosphatidylcholine for 24 hours.Fluvastatin time groups:HUASMC were incubated with the same dose of fluvastatin for 1 hour before incubated with lysophosphatidylcholine for 6 h,12 h,24 h,48 h,72 h. The cell proliferation evaluated by cell counting and MTT assay,the cell cycle examined with Flow Cytometry,the apoptosis of HUASMC examined with dying Hoechst33258,the expression of Survivin and Fas detected in immunohistochemical methods,the mRNA expression of Survivin and Fas determined by reverse transcription polymerase chain reaction method(RT-PCR).Results:(1)The proliferation and cell counting of vascular smooth muscle cells in LPC group was significantly increased compared with the control group,fluvastatin could inhibit the proliferation of LPC-induced HUASMC in a dose dependent manner.(2)Flow Cytometric DNA revealed that more vascular smooth muscle cells in LPC group got into the S or G2/M phase,and the apoptosis rate was makedly decreased as compared with the control.Fluvastatin could induce significantly enhancement at G0/G1 phase and arrenuation at S phase vascular smooth cell compared with LPC group.In a dose dependent manner the apoptosis rate increased makedly.(3)Hoechst-33258 revealed that apoptosis cells was increased with the time elapsing and the dose of fluvastatin increasing.(4)The LPC could significantly stimulate the mRNA expression of Survivin compared with the control group. Immunocytochemistry revealed that there were many brown particles in cells.Fluvastatin could inhibit the mRNA expression of Survivin in the early period,on the contrary,with the time elapsing,the mRNA expression of Fas become more and more strongly,at 72 h,the mRNA expression of Fas reach the peak.Conclusion:Fluvastatin could inhibit proliferation on HUASMC induced by lysophosphatidylcholine(LPC).Fluvastatin could induce apoptosis associated with induction of Survivin,Fas and play a significant role down-regulation of Survivin and up-regulation of Fas on HUASMC.