眼镜蛇毒L-氨基酸氧化酶分离纯化及对血管内皮细胞的抑制作用  

Purification of L-amino acid oxidase from Naja atra venom and its effects on endothelial cells

作　　者：陈洲[1] 黄剑钧[1] 薛玲[2] 许云禄[1] 林丽珊[3] 

机构地区：[1]福建医科大学药学院,福建福州350004 [2]福建医科大学基础医学院,福建福州350004 [3]莆田学院药学系,福建莆田351100

出　　处：《中国药理学通报》2009年第8期1024-1028,共5页Chinese Pharmacological Bulletin

基　　金：福建省科技厅资助省属高校资助项目(No2006F5049);莆田市科技计划资助项目(No2005S07)

摘　　要：目的从眼镜蛇毒中分离纯化L-氨基酸氧化酶(L-a-mino acid oxidase of Naja atra venom,NAV-LAAO),观察其对血管内皮细胞是否有抑制作用。方法应用阳离子交换层析和亲和层析方法分离纯化NAV-LAAO;采用MTT法、Western blot测定caspase及细胞小管形成实验观察NAV-LAAO对人脐静脉内皮细胞(HUVEC)生长和血管生成的影响。结果眼镜蛇毒粗毒经两步层析法得到的LAAO,分子量约为58 ku。NAV-LAAO抑制HUVEC细胞的生长和细胞小管形成,其抑制细胞生长的IC50为21.42 mg.L-1。与对照组相比,LAAO组caspase-3、caspase-8升高。结论应用两步层析法从眼镜蛇毒中分离出的NAV-LAAO具有剂量依赖性的抑制HUVEC细胞的生长和影响细胞小管形成的作用。Aim To purify L-amino acid oxidase （LAAO） from the venom of Naja atra and study its effect on endothelial cells. Methods The NAV-LAAO was purified by ion-exchange chromatography and affinity chromatography. The MTT assay and Western blot were used to detect the viability and apoptosis of HUVEC. The tubule-forming was used to study the angiogenesis of cells. Results The NAV-LAAO was purified successfully from the venom of Naja atra. The molecular weight of NAV-LAAO was determined to be 58 ku by SDS-PAGE. NAV-LAAO effectively inhibited the growth and tubule-forming of HUVEC, and the 50% inhibitory concentration （IC50） was 21.42 mg·L^-1. Compared with control, the levels of caspase-3 and caspase-8 increased in HUVEC treated with NAV- LAAO. Conclusions The NAV-LAAO is purified successfully from Naja atra venom by ionexchange chromatography and affinity chromatography. The NAV- LAAO inhibits the growth and tubule-forming capacity of HUVEC in a dose-dependent manner.

关 键 词：眼镜蛇毒 L氨-基酸氧化酶 分离纯化 血管内皮细胞 内皮/细胞小管形成 亲和层析 

分 类 号：R322.123[医药卫生—人体解剖和组织胚胎学] R329.2[医药卫生—基础医学]