论文部分内容阅读
目的观察C反应蛋白(CRP)对体外培养人脐静脉内皮细胞(HUVEC)凋亡相关基因及蛋白Bcl-2／Bax表达的影响。方法采用酶消化法培养人脐静脉内皮细胞,将CRP(浓度为0、1、5、25 mg／L)与内皮细胞作用24 h后通过流式细胞仪测定凋亡细胞比率。选用25 mg／L CRP作用内皮细胞(0、4、8 h),采用逆转录-聚合酶链反应(RT-PCR)、Western-Blotting方法观察凋亡相关基因及蛋白Bcl-2／Bax表达的变化。结果随CRP浓度的增加诱导内皮细胞凋亡的比率逐渐增加,与对照组(5．42±0．92)％相比,25 mg／L时细胞凋亡率达(15．71±2．34)％,显著增加(P<0．01);内皮细胞在25 mg／L CRP作用4、8 h后Bax mRNA、蛋白表达逐渐增强,Bcl-2 mRNA、蛋白表达逐渐减弱,在8 h它们表达量与对照组相比差异有统计学意义(P<0．05)。结论临床相关浓度的CRP可通过调节Bcl- 2／Bax mRNA、蛋白的表达诱导内皮细胞凋亡。
Objective To investigate the effect of C-reactive protein (CRP) on the expression of Bcl-2 / Bax protein and apoptosis related genes in cultured human umbilical vein endothelial cells (HUVECs). Methods Cultured human umbilical vein endothelial cells with enzyme digestion method, the ratio of apoptotic cells was determined by flow cytometry after CRP (0,1,5,25 mg / L) was incubated with endothelial cells for 24 h. The effects of 25 mg / L CRP on endothelial cells (0, 4 and 8 h) were observed. The expressions of Bcl-2 / Bax and apoptosis related genes were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western- Variety. Results The percentage of apoptosis induced by CRP increased gradually. Compared with the control group (5.42 ± 0.92)%, the apoptosis rate reached 25.77 ± 2.34 ), And significantly increased (P <0.01). The expression of Bax mRNA and protein in endothelial cells gradually increased after treated with 25 mg / L CRP for 4 and 8 h, while the expression of Bcl-2 mRNA and protein decreased gradually Compared with the control group, the difference was statistically significant (P <0.05). Conclusion Clinically relevant concentrations of CRP can induce apoptosis of endothelial cells by regulating the expression of Bcl-2 / Bax mRNA and protein.