氟伐他汀对溶血性磷脂酰胆碱致动脉平滑肌细胞增殖和凋亡相关因子表达的影响

Effect of fluvastatin on expression of apoptosis correlation factor and proliferation induced by lysophosphatidylcholine in human umbilical artery vascular smooth muscle cells

目的:观察氟伐他汀对溶血性磷脂酰胆碱(LPC)诱导人脐动脉平滑肌细胞(HUASMC)增殖和凋亡作用的影响,探讨他汀类药物抗动脉粥样硬化的机制。方法:采用体外培养的HUASMC,待其生长到融合状态时用不同浓度(10-7～10-5mol/L)氟伐他汀和LPC(1、2.5、5mg/L)干预后用MTT法检测细胞增殖,Hoechst-33258染色法观察细胞凋亡,免疫组化染色方法和RT-PCR检测血管平滑肌凋亡相关因子Survivin、Fas的表达。结果:(1)MTT结果反映LPC作用24hHUASMC数量明显增多,增殖指数增大,氟伐他汀能抑制LPC促增殖的作用,使OD值降低,细胞数量明显减少;(2)免疫组化和RT-PCR结果显示:LPC对照组(5mg/L)和氟伐他汀处理组在作用早期(6～12h)Survivin因子表达明显,胞浆内可见大量棕黄色颗粒,随时间延长Survivin因子表达逐渐下降,72h时Survivin因子表达明显减弱,与LPC对照组比较差异有统计学意义(P<0.05);而LPC对照组和氟伐他汀处理组在早期(6～24h)Fas因子表达不明显,晚期(48～72h)可检测到Fas因子大量表达,胞浆内可见棕黄色颗粒,晚期处理组与LPC对照组比较差异有统计学意义(P<0.01)。结论:氟伐他汀抑制LPC对HUASMC的促增殖作用并诱导细胞发生凋亡,其凋亡作用主要与抑制Survivin因子和促进Fas因子表达有关。

To observe effects of fluvastatin and lysophosphatidylcholine （LPC） on the proliferation and apoptosis of human umbilical artery vascular smooth muscle cells （HUASMC） and provide a new evidence to statius in clinical application regarding antiatherosclerotic action. METHODS： Proliferation in cultured VSMC, derived from HUASMC and treated with different doses of fluvastatin and LPC, was analyzed by means of MTI＇ assay. Apoptosis of the cell was examined using dying Hoechst33258. Expression of Survivin and Fas were examined using immunocytochemistry and reverse transcription polymerase chain reaction method. RESULTS： （1） LPC （5 mg/L） could increase index of proliferation significantly. Compared with LPC group, fluvastatin could decrease proliferation significantly. （2） Immunocytochemistry and reverse transcription polymerase chain reaction method revealed that the apoptesis process was associated with inhibition of Survivin and activation of Fas. CONCLUSION： Fluvastatin inhibits proliferation of HUASMC induced by LPC. Fluvastatin can induce apoptosis of HUASMC in connection with down-regulation of Survivin and up-regulation of Fas.