【摘要】 目的研究眼镜蛇毒细胞毒素13诱导人脐静脉内皮细胞凋亡作用及其对Bcl-2家族基因表达的影响。方法CCK-8法测定细胞毒素13对人脐静脉内皮细胞增殖的影响;Annexin V-FITC/PI双染法观察细胞凋亡形态学改变;流式细胞术分析DNA倍体及细胞凋亡率;逆转录聚合酶链反应检测Bcl-2、Bax和Bcl-xL基因表达的变化,Western blotting检测Bcl-2和Bax蛋白表达的变化。结果细胞毒素13对人脐静脉内皮细胞增殖具有显著的抑制作用,并可诱导其发生凋亡,其抑制作用呈剂量依赖性。细胞毒素13可上调促凋亡蛋白Bax,而对抗凋亡蛋白Bcl-2和Bcl-xL的表达无明显影响。结论细胞毒素13可能通过上调Bax表达诱导细胞凋亡,抑制人脐静脉内皮细胞增殖。更多还原

【Abstract】 Aim To investigate the effects of cobra cardiotoxin-13 (CTX-13) on the apoptosis of human umbilical vein endothelial cells (HUVEC) and the expression of Bcl-2 family.Methods The effect of CTX-13 on HUVEC proliferation was analyzed by CCK-8 assay.The morphological changes were observed with Annexin V-FITC/PI staining.The DNA ploidy analysis and apoptotic rates assay were performed by flow cytometry.The expression level of Bcl-2 gene family was measured by RT-PCR and Western blotting.Results CTX-13 caused a dose-dependent decrease of cell viability and displayed several features of apoptosis including translocation of membrane phosphatidylserine and increase of sub G1 hypodiploid cell population.The apoptotic rates of HUVEC treated with CTX-13 concentrations of 0.3,0.75,1.5 μmol/L were 20.50%±2.62%,54.20%±4.93% and 93.40%±6.45%.CTX-13 up-regulated the mRNA expression of Bax about 4-fold as well as its protein expression,but did not affect the expression of Bcl-2 and Bcl-xL.Conclusions CTX-13 suppressed HUVEC proliferation significantly and induced apoptosis in a dose-dependent manner.CTX-13 up-regulated Bax in mRNA and protein level.It was probable the CTX-13 suppressed proliferation and induced apoptosis by up-regulating Bax.更多还原