大鼠肝微粒体谷胱甘肽S-转移酶简易制备法对活性影响  

Augmentation of rat liver microsome glutathione S-transferase preparation

作　　者：郑英[1] 张捷[1] 楼宜嘉[1] 

机构地区：[1]浙江大学药学院,浙江杭州310031

出　　处：《浙江大学学报（医学版）》2002年第6期429-432,共4页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然基金资助项目 (30 0 70 90 4 )

摘　　要：目的 :研究大鼠肝微粒体简易制备法对微粒体谷胱甘肽巯基转移酶 (m GST)活性的影响。方法 :用聚乙二醇 6 0 0 0 (PEG 6 0 0 0 )法和钙沉淀法制备大鼠肝微粒体 ,并与巯基烷化剂 N-乙基顺丁烯二酰亚胺 (NEM)共孵育 ,分别测定不同制备法所获 m GST活性。结果 :PEG 6 0 0 0法和钙沉淀法制备的肝微粒体 ,其 m GST活性分别为0 .15 ,0 .0 8μmol· min- 1 · ml- 1 ;NEM可使其活性分别显著增加至 1.797和 2 .375倍 (P<0 .0 0 1) ;洗涤纯化后 ,分别增加至 NEM活化前的 2 .35 0和 4 .12 7倍 (P<0 .0 0 1) ;其中 NEM与微粒体共孵育最适浓度为 5～ 10 mmol/ L ,最佳激活时间为 1～ 2 min。结论 :经 PEG 6 0 0 0法和钙沉淀法高速离心制备的微粒体 ,分离制备时间短 ,方法简单易行 ;钙沉淀法制备的 mObjective: To investigate the augmentation of microsomal glutathione S transferase (mGST) preparation using simple organic compounds. Methods: Rat liver microsomes were isolated using both the polyethyleneGlyco 6000 (PEG6000) and Ca 2+ precipitation methods. Next the mGST activity was measured after incubation with the alkylating agent N ethylmaleimind (NEM). Results: The baseline mGST activity of rat microsomes was 0.15 after PEG6000 exposure and 0.082 after Ca 2+ precipitation. After NEM treatment mGST activity increased to 1.797 (2.35×) ( P <0.01) and 2.375 (4.127×) ( P <0.01) respectively followed by purified wa shing. mGST activation was stimulated maximally by 5 10 mmol/L NEM and occurred rapidly with 1 2 min of co incubation . Conclusion: For both the PEG6000 and Ca 2+ precipitation methods the mGST activity of rat microsomes can be significantly enhanced after exposure to NEM. This enhancement is more prominent with the Ca 2+ precipitation.

关 键 词：肝微粒体 酶学 谷胱甘肽转移酶 代谢 聚乙二醇6000法 钙沉淀法 N-乙基顺丁烯二酰亚胺 

分 类 号：R962[医药卫生—药理学]