眼镜蛇毒金属蛋白酶atrase A诱导人内皮细胞释放炎症介质及凋亡  

Inflammatory mediators releasing and apoptosis of endothelial cell induced by cobra venom metalloproteinase atrase A

作　　者：叶巧玲[1] 孙黔云[1] 李敏[2] 

机构地区：[1]贵州省中国科学院天然产物化学重点实验室 [2]贵州省人民医院呼吸内科,贵州贵阳550002

出　　处：《中国药理学通报》2009年第8期1001-1006,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30560035);贵州省科技攻关项目(No2002-1075);贵州省优秀青年科技人才项目(No2005-0510);贵州省优秀科技教育人才省长专项资金资助项目

摘　　要：目的研究眼镜蛇毒金属蛋白酶atrase A对人内皮细胞的作用。方法采用MTT法、SRB法、形态学观察分别检测atrase A对人微血管内皮细胞生长的影响;内皮细胞经at-rase A处理后,ELISA法检测内皮细胞释放IL-8、ICAM-1、MCP-1、E-selectin的变化,萤光发光法检测各组caspase-8、caspase-3/7的表达情况。经尾静脉给予大鼠atrase A后,检测大鼠血清中ICAM-1、IL-8、TNF-α的变化情况。结果At-rase A(400、40 mg.L-1)对内皮细胞的生长具有明显的抑制作用。而MTT法结果显示,atrase A对高接种密度的内皮细胞基本上没有影响;镜检结果显示,atrase A(400、40 mg.L-1)可使贴壁生长的内皮细胞解离悬浮。而4 mg.L-1的atrase A对内皮细胞的生长没有影响。高剂量atrase A(400mg.L-1)可诱导内皮细胞IL-8、ICAM-1、MCP-1释放增加,而低剂量atrase A(4 mg.L-1)对各炎症介质的表达没有影响。Atrase A还可诱导内皮细胞caspase-8、caspase-3/7的表达,12 h达峰值。经尾静脉分别给予2 240μg.kg-1和400μg.kg-1的atrase A后,高剂量组SD大鼠血清中ICAM-1、IL-8、TNF-α表达增加;而低剂量组中没有明显变化。结论眼镜蛇毒金属蛋白酶atrase A可抑制内皮细胞生长,诱导内皮细胞释放炎症介质和凋亡。Aim To investigate the effect of cobra venom metalloproteinase atrase A on human endothelial cell. Methods The effects of atrase A on the growth of HMEC were measured by MTT, SRB and morphological observation methods, respectively. After exposure to atrase A, IL-8, ICAM-1, MCP-1 and E-selectin released by HMEC were detected. Caspase-8 and caspase-3/7 were detected by fluorescent luminescence method. After intravenous injected atrase A to rats, the concentrations of ICAM-1, IL-8 and TNF-α in serum were measured. Results The results showed that atrase A （400, 40 mg·L^-1） significantly inhibited the growth of HMEC in low cell density. The microscopic examination revealed that atrase A detached the adherent HMEC. After exposure to atrase A, IL-8, ICAM-1 and MCP-1 released by HMEC were increased. Atrase A induced HMEC to express caspase-3/7 and caspase-8. After the administration of atrase A to rats, the concentrations of ICAM-1, IL-8 and TNF-α in serum were increased. There was no difference between the low-dose group and control group in our experiments. Conclusion Cobra venom metalloproteinase atrase A inhibits the growth of HMEC, and induces HMEC releasing inflammation mediators and apoptosis.

关 键 词：蛇毒金属蛋白酶 内皮细胞 炎症 凋亡 炎症因子 细胞粘附分子 眼镜蛇毒 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]