五味子乙素抑制M146L细胞Aβ_(42)生成的机制研究  

γ-Schisandrin inhibits production of amyloid β-protein 42 in M146L cells

作　　者：刘薇[1] 余锐[1] 吴家华[1] 罗焕敏[1] 

机构地区：[1]暨南大学药学院神经药理研究室,广东广州510632

出　　处：《药学学报》2006年第12期1136-1140,共5页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(30472014).

摘　　要：目的研究五味子乙素抑制M146L细胞A4β2生成的机制。方法体外培养高效表达Aβ42的M146L细胞株,分别加入不同浓度的五味子乙素(1.67,5.00和15.00μg.mL-1)、β分泌酶抑制剂(S4562,100.00μg.mL-1)和γ分泌酶抑制剂(S2188,13.68μg.mL-1)。用CCK-8(cell counting k it-8)比色法检测不同处理对M146L细胞活性的影响;用ELISA法测定M146L细胞所分泌的Aβ42的变化;用W estern b lotting检测APP的β分泌酶剪切产物C99蛋白的含量变化,结合用β和γ分泌酶活性试剂盒,检测五味子乙素对这两种酶活性的影响。结果不同处理因素对M146L细胞的存活率均无影响,不具有细胞毒作用。中、高剂量的五味子乙素均不同程度地抑制M146L细胞分泌A4β2及γ分泌酶的活性,但都不改变M146L细胞C99蛋白的含量及β分泌酶的活性。结论五味子乙素可抑制γ分泌酶活性,其降低A4β2生成是通过抑制γ分泌酶的活性来实现的。Aim To investigate the inhibition of amyloidβ-protein 42 （Aβ42） production in M146L cells by γ-schisandrin. Methods M146L cells which can produce considerable Aβ42 in vitro were treated with γ-schisandrin （ 1.67, 5.00 and 15.00 μg · mL^-1 ） , β-secretase inhibitor （ S4562, 100.00 μg ·mL^-1） and γ-secretase inhibitor （S2188, 13.68 μg · mL^-1）, separately. Cell counting kit-8 （CCK-8） was used to assess cell viability. Enzyme-linked immunosorbent assay （ELISA） was carried out to determine the amount of Aβ42. Western blotting was used to examine C99, an intermediary product of APP cleaved by β-secretase. β-Secretase and γ-secretase activities were assayed by commercial kits. Results The CCK-8 assay indicated that different concentrations of γ-schisandrin had no neurotoxicity on the cultured M146L. And the ELISA test showed that the amount of Aβ42 secreted by M146L ceils treated with γschisandrin （5.00 and 15.00 μg · mL^-1） decreased obviously as compared with solvent control. The results of Western blotting test indicated that there was no change of C99 contents and β-secretase activity in β-schisandrin treated ceils, while γ-secretase activity decreased obviously. Conclusion γ-Schisandrin inhibited production of Aβ42 in M146L ceils through inhibiting γ-secretase.

关 键 词：五味子乙素 阿尔茨海默病 口淀粉样蛋白 分泌酶抑制剂 

分 类 号：R965.1[医药卫生—药理学] R971.91[医药卫生—药学]