目的探讨广东海风藤多糖对M146L细胞分泌的β淀粉样蛋白的影响。方法体外培养稳定转染人类阿尔茨海默病(AD)β淀粉样前体蛋白(APP)基因及突变型早老素1(PS1)基因的CHO细胞系M146L,使之高效产生β淀粉样蛋白42(Aβ42),建立Aβ42过度表达的细胞模型。加入待筛选的药物广东海风藤多糖,用四唑盐(MTT)比色法检测不同浓度的广东海风藤多糖(0.3125、0.625、1.25μg/ml)对M146L细胞的毒性作用,应用酶联免疫法(ELISA)观察细胞分泌的Aβ42的变化。结果不同浓度的广东海风藤多糖对M146L存活率没有影响,不具有细胞毒作用。0.625、1.25μg/ml的广东海风藤多糖对M146L细胞分泌Aβ42有抑制作用,呈剂量依赖性。结论一定剂量的广东海风藤多糖对M146L细胞分泌的Aβ42有明显的抑制作用,其机制有待进一步研究。
Objective To study the inhibition of amyloid β-protein production in M146L cell by Kadsura heteroclita polysaccharide. Methods Aβ42 overexpressing cell model in vitro was established by culturing Chinese Hamster Ovary Cells stably expressing amyloid beta-protein precursor and mutant presenilin M146L. The MTT assay was used to test cytotoxicity of Kadsura heteroelita polysaccharide, and ELISA was carried out to analyze Aβ42 production. Results The MTF assay showed that Kadsura heteroclita polysaccharide at different concentrations didn't have any toxic effect on M146L cell survival. Kadsura heteroclita polysaccharide at the concentration of 0. 625 and 1.25 μg/ml significantly inhibited the production of Aβ42. Conclusions The suitable dose of Kadsura heteroclita polysaccharide can inhibite the product of Aβ42 in M146L cell, and its mechanism needs further study.