中华眼镜蛇毒活性组分诱导内皮细胞凋亡的机制  

The mechanism of China cobra venom active factor for inducing apoptosis in endothelial cells

作　　者：刘新艳[1] 余清声[1] 王桂平[2] 余红娥[3] 朱柳[1] 袁牧[1] 楼晓华[4] 腾脉坤[4] 

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]广东药学院中山校区,广东中山528458 [4]中国科学技术大学生命科学院,安徽合肥230026

出　　处：《中国生化药物杂志》2011年第4期257-260,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：广州市教育局市属高校科技计划项目(NO2004-1016)

摘　　要：目的通过检测内皮细胞Bcl-2和caspase3的表达水平,探讨中华眼镜蛇毒活性组分(CCVAF)诱导内皮细胞凋亡的可能机制。方法采用血管内消化法原代培养牛肺主动脉血管内皮细胞(BAVEC)。实验分为不同浓度的CCVAF组(1.25,2.5,5,10 mg/L)和对照组(等体积的培养基)。采用流式细胞仪检测CCVAF作用24 h后各组BAVEC的Bcl-2表达水平;采用比色法测定caspase3的活性。结果经CCVAF作用后,流式细胞仪测定各组均未见到有阳性峰出现在M1,抗凋亡蛋白Bcl-2表达并没有受到药物的影响。而与对照组相比,capase3活性在1.25mg/L组即出现升高,此后随浓度的增加而升高,呈剂量依赖性(r=0.929,P<0.05)。各浓度组加入capase3活性抑制剂(Ac-DEVD-CHO)后,capase3活性明显降低,但仍然高于对照组(P<0.05)。结论 CCVAF不影响内皮细胞Bcl-2的表达,而能够上调capase3的活性。其可能不经Bcl-2影响的线粒体所在的凋亡通路调节细胞的凋亡。Purpose To investigate the mechanism of China cobra venom active factor（CCVAF） for inducing apoptosis in endothelial cells by monitoring the expression level of Bcl-2 and caspase3 in endothelial cells.Methods Bovine arteria pulmonalis vascular endothelial cells（BAVEC） were cultured in primary.The cells were divided into different concentrations of CCVAF group（1.25,2.5,5,10 mg/L） and control group（isovolumic medium）.The expression of correlated protein Bcl-2 of BAVEC in each group was detected with flow cytometer and caspase3 activity was detected by chromatometry after CCVAF treatment for 4 h.Results Four hours after treatment with CCVAF,it was confirmed that no effect was found that on the expression of protein Bcl-2 at a series of concentrations（1.25,2.5,5,10 mg/L）and in control group.M1 positive peak didn′t appear in all groups.On the other hand,caspase3 activity was elevated at 1.25 mg/L group,and showed a dose-dependent manner（r=0.929,P0.05）.Compared with control group there was significant difference（P0.05）.When a specific inhibitor of caspase3 activity（Ac-DEVD-CHO） was included in the different groups,the protease activity significantly depressed but increased compared with control group（P0.05）.Conclusion CCVAF can up-regulate activity of caspase3,but there is no significant relationship with expression of anti-apoptotic protein Bcl-2.These findings indicate that CCVAF does not lead to apoptosis of endothelial cells by regulating apoptosis pathway including chondrosome of Bcl-2 influence.

关 键 词：眼镜蛇毒 内皮细胞 BCL-2 capase3 凋亡 血管生成 肿瘤 

分 类 号：R285.5[医药卫生—中药学]