中华眼镜蛇毒活性组分诱导内皮细胞凋亡  

China cobra venom active factor can induce apoptosis of endothelial cells

作　　者：刘新艳[1] 余清声[1] 余红娥[1] 朱柳[1] 王桂平[2] 袁牧[1] 楼晓华[3] 腾脉坤[3] 

机构地区：[1]广州医学院药物研究中心,广东广州510182 [2]广州医学院护理学院,广东广州510182 [3]中国科学技术大学生命科学院,安徽合肥230026

出　　处：《中国药理学通报》2008年第6期822-826,共5页Chinese Pharmacological Bulletin

基　　金：广东省自然科学基金资助项目(No980463);卫生部科技基金资助项目(No98-2-349)

摘　　要：目的观察中华眼镜蛇毒活性组分(China cobra venom active factor,CCVAF)诱导内皮细胞凋亡的作用。方法采用荧光显微镜、流式细胞仪等方法检测CCVAF对牛肺主动脉内皮细胞(bovine arteria pulm onalis vascular endothelialcells,BAVEC)凋亡形态学、DNA、细胞周期以及凋亡指数的影响。结果经AO染色、流式细胞仪分析均证实CCVAF能够诱导内皮细胞出现凋亡。荧光显微镜下观察到CCVAF各浓度(0.15625、0.3125、0.625、1.25μmol.L-1)组处理的内皮细胞,核染色质固缩或断裂成片段状,染色不均匀,出现凋亡小体,凋亡细胞的比例随浓度升高而增加,0.625μmol.L-1组视野内凋亡细胞最多;流式细胞仪分析结果表明CCVAF在浓度低于0.625μmol.L-1,细胞生长被阻滞在S期,1.25μmol.L-1时则被阻滞在G0/G1期,且细胞凋亡率随浓度升高而逐渐增加。结论CCVAF能够诱导内皮细胞出现凋亡,通过此途径抑制内皮细胞的生长增殖,这可能是其对抗血管生成的机制。Aim To discuss the apoptotic effect of bovine arteria pulmonalis vascular endothelial ceils （BAVEC） induced by China cobra venom active factor （CCVAF）. Methods Morphology of apoptotic BAVEC was observed by fluorescence microscope, and cell cycle and apoptotic index were dectected by flow cytometry technique. Results The present study indicated that CCVAF could induce effectively BAVEC to apoptosis. After BAVEC were treated with different concentrations of CCVAF （ 0. 15 625,0. 3125,0. 625 and 1.25 μmol·L^-1）, apoptotic morphology of BAVEC including nuclear chromatin pyknosis or breakage, staining uneven and apoptotic body etc,were observed under fluorescence microscope. CCVAF in-duced the apoptosis of BAVEC in a dose-dependent manner,the apoptotic peak was observed in the dose of 0. 625 μmol·L^-1. The results of flow cytometry study showed that most BAVEC were blocked in S stage of cell cycle when the dose of CCVAF was under 0. 625 μmol·L^-1, however blocked in G0/G1 stage in 1.25 μmol·L^-1 CCVAF, and the apoptotic rate increased in a dose-dependent manner. Conclusions CCVAF could induce the apoptosis of BAVEC,which might be the important mechanism that CCVAF inhibited the proliferation of BAVEC and angiogenesis.

关 键 词：眼镜蛇毒 内皮细胞 细胞凋亡 血管生成 肿瘤 

分 类 号：R-332[医药卫生] R329.25