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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 2 4～48h,换成含 0 .5 m mol/ L SNAP的条件培养液 ,8～ 10 h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 Bcl- 2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。
Objective To quantitatively detect the expression of Bcl-2 protein in vascular smooth muscle cells (VSMC) by nitric oxide (NO), and further investigate the effect of bcl-2 overexpression on the apoptosis of VSMC induced by nitric oxide (NO) Role in NO-induced VSMC Apoptosis. Methods SNAP was used as NO donor. Rat VSMCs from 5 to 8 passages were inoculated into Petri dishes. The conditioned medium was incubated for 24-48 h and replaced with 0.5-5 mol / L SNAP conditioned medium. After 8 to 10 hours, the cells were fixed and the antigens were detected by indirect immunofluorescence. The expression of Bcl-2 protein in the cells was quantitatively detected by the adherent cytometer. The bcl-2 gene was introduced into VSMC by retroviral vector and overexpressed. The apoptosis rate of SNAP-treated cells was detected and compared with untransfected VSMC. Results SNAP decreased the expression of Bcl-2 protein in VSMC, while overexpression of bcl-2 inhibited the apoptosis of VSMC induced by SNAP. Conclusion Down-regulation of bcl-2 expression is one of the ways of SNAP-induced VSMC apoptosis.