C反应蛋白调节bcl-2/bax蛋白表达诱导人脐静脉内皮细胞凋亡

REVULSIVE EFFECT OF C-REACTIVE PROTEIN ON THE APOPTOSIS OF HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS THROUGH REGULATING THE EXPRESSION OF BCL-2/BAX PROTEIN

目的观察不同浓度C反应蛋白对体外培养的人脐静脉内皮细胞(Human umbilical vein endothelial cells,hUVEC)凋亡的影响及其分子机制。方法体外培养人脐静脉内皮细胞,用吖啶橙/溴化已啶荧光染色观察凋亡细胞形态。四甲基偶氮唑蓝比色法(MTT)分别测定与不同浓度C反应蛋白(5mg/L、10mg/L、20mg/L)作用12、24、48h后hUVEC的增殖率,流式细胞仪和Western blotting分别检测与不同浓度C反应蛋白作用24h后的细胞早期凋亡率及bcl-2/bax蛋白表达水平。结果随着C反应蛋白浓度的增加,hUVEC的MTT值逐渐降低,而细胞早期凋亡率则逐渐升高。与对照组相比,24h后高C反应蛋白组MTT值明显下降(P<0.05),而细胞早期凋亡率显著增加(P<0.01)。内皮细胞在高C反应蛋白作用24h后,bcl-2蛋白表达减弱,bax蛋白表达增强,它们的表达量与对照组相比差异有统计学意义(P<0.05)。结论 C反应蛋白可能通过调节bcl-2/bax蛋白表达诱导人内皮细胞凋亡。

Objective To study the effects of C-reactive protein（CRP） on the apoptosis of human umbilical vein endothelial cells（hUVEC） and to explore its potential molecular mechanism.Methods The hUVECs were cultured in vitro.Apoptosis of HUVEC were observed by acridine orange/ethidium bromide fluorescence staining.The survival rates of hUVECs were detected by MTT assay after hUVECs were incubated with different CRP concentrations（5mg/L、10mg/L、20mg/L）for 12、24、48 hours.The early apoptosis rates and the bcl-2/bax protein expression were detected respectively by flow cytometry and western blotting after hUVECs were incubated with different CRP concentrations for 24 hours.Results With the increase in CRP concentration,the value of MTT was significantly reduced,while the early apoptosis rates were gradually increased.High CRP concentrations inhibited the expression of bcl-2 protein and promoted the expression of bax protein.Conclusion High glucose may induce the apoptosis of hUVEC through regulating the expression of bcl-2/bax protein.