bcl—2在一氧化氮诱导血管平滑肌细胞凋亡中的作用

目的：对一氧化氮（NO）作用下血管平滑肌细胞（VSMC）中bcl-2蛋白表达的变化进行定量检测，并进一步探讨bcl-2过表达对NO诱导VSMC凋亡的影响，以确定bcl-2在NO诱导VSMC凋亡中的作用。方法以亚硝基乙酰青霉胺（SNAP）作为NO供体，将5－8代的大鼠VSMC接种于Petri培养皿中，条件培养液培养24－48h，换成含0．5mmol/LSNAP的条件培养液8－10h后，固定细胞、间接免疫荧光法检测抗原，借助粘附式细胞仪对细胞中bcl-2蛋白表达进行定量检测，通过逆转录病毒载体将bcl-2基因导入VSMC，使其过量表达，检测SNAP作用下细胞的凋亡率，并与未经转染的VSMC进行比较，结果：SNAP使得VSMC中B蛋白表达下调，而bcl-2过表达能够抑制SNAP作用下VSMC的凋亡。结论下调bcl-2表达是SNAP诱导VSMC凋亡的途径之一。

ROLE OF bcl-2 IN APOPTOSIS OF VASCULAR SMOOTH MUSCLE CELLS INDUCED BY NITRIC OXIDE

Objective To explore the effect of bcl 2 in vascular smooth muscle cells(VSMC) apoptosis induced by nitric oxide(NO),we investigated the effect of NO on Bcl 2 protein expression in cultured vascular smooth muscle cells(VSMCs),and effect of Bcl 2 over expression on apoptosis is(VSMCs) induced by NO. Methods Cultured rat VSMCs at passage 5 to 8 were used for experiments,S nitroso N acetypenicillamine(SNAP) was used as NO donor.Cells were preincubated for 24-48 hours in special culture dish with conditioned medium to reach quiescent,and then incubated in conditioned medium containing 0 5 mmol/L SNAP.After 8 to 10 hours,the cells were fixed and Bcl 2 expression was analyzed by ACAS 570 through fluorescent immunochemistry.Vascular smooth muscle cells were transfected with bcl 2 gene through retrovirus to allow overexpression of Bcl 2 in VSMCs.Finally,we observed the apoptosis in normal VSMCs and VSMCs infected with bcl 2. Results SNAP downregulated Bcl 2 expression in VSMCs.PCR and Southern hybridization confirmed the integration of the bcl 2 retrovirus vector into vascular smooth muscle cells genomic DNA.And fluorescence immunohistochemistry confirmed that Bcl 2 protein was overexpressed in VSMCs infected with bcl 2.And ratio of apoptosis become lower in VSMCs infected with bcl 2. Conclusion bcl 2 can inhibit apoptosis of VSMCs induced by SNAP.It is suggested that downregulation of Bcl 2 protein is one of the ways in which SNAP induces VSMCs apoptosis,and apoptosis in VSMCs induced by SNAP is regulated by bcl 2.;