辛伐他汀对脓毒症大鼠血清诱导的人脐静脉内皮细胞凋亡的影响

The effects of simvastatin on the apoptosis of human umbilical vein endothelial cells induced by serum of sepsis rats

目的初步探讨辛伐他汀对脓毒症大鼠血清诱导的人脐静脉内皮细胞凋亡的影响及可能机制。方法采用改良盲肠结扎穿孔术复制脓毒症模型,收集血清备用。体外培养的人脐静脉内皮细胞株(ECV-304)分为三组(对照组、脓毒症组、辛伐他汀干预组),培养24 h,MTT法检测细胞活性,AO染色观察细胞凋亡情况,RT-PCR检测细胞Bcl-2 mRNA、Bax mRNA表达。结果 MTT:与脓毒症组相比,辛伐他汀干预组细胞活性明显提高;AO染色:对照组细胞形态基本正常,脓毒症组可见大量凋亡细胞,辛伐他汀干预组可见少量细胞凋亡;RT-PCR:与对照组相比,脓毒症组、辛伐他汀干预组,Bcl-2 mRNA、Bax mRNA高表达(P<0.01);与脓毒症组相比,辛伐他汀干预组Bcl-2 mRNA高表达(P<0.05),Bax mRNA低表达(P<0.01)。结论辛伐他汀可抑制脓毒症大鼠血清诱导的内皮细胞凋亡,上调Bcl-2和下调Bax表达可能是其机制之一。

Objective To investigate the effects of simvastatin on the apoptosis of human umbilical vein endothelial cells induced by serum of sepsis rats and to explore its potential molecular mechanism.Methods Adopting the modified cecal ligation and puncture to replicate the sepsis animal model and collect rats serum for experiments.Human umbilical vein endothelial cells（ECV-304） cultivated with DMEM medium in vitro,and were divided into 3 groups（control group,sepsis group,simvastatin intervention group）.MTT colorimetry were performed to confirm cell apoptotic ratio,acridine orange staining was used to observe the cell apoptosis morphologic changes,RT-PCR method was used to detect the expression of mRNA of Bcl-2 and Bax.Results MTT： Compared with the sepsis group,activity of ECV-304 cells in simvastatin intervention group increased.AO staining： the control group cell morphology was normal,sepsis group showed large amounts of apoptotic cells,and simvastatin intervention group with small quantities of apoptosis.RT-PCR：Compared with the control group,Bcl-2 mRNA and Bax mRNA expression levels of the sepsis and simvastatin intervention group were up-regulated（P＆lt;0.01）.Compared with the sepsis group,Bcl-2 mRNA expression levels of simvastatin intervention group were up-regulated（P＆lt;0.05）,while Bax mRNA expression levels were down-regulated（P＆lt;0.01）.Conclusion Simvastatin can reduce the number of apoptosis of ECV-304 cells induced by serum from sepsis rats,the underlying mechanism might be related to rise expression of Bcl-2 gene and downregulation of Bax expression.