黄芪注射液及其有效成分对乳腺癌细胞增殖和Akt磷酸化的影响  

Effects of Astragalus injection and its ingredients on proliferation and Akt phosphorylation of breast cancer cell lines

作　　者：邓樱[1] 陈红风[1] 

机构地区：[1]上海中医药大学龙华医院乳腺科,上海200032

出　　处：《中西医结合学报》2009年第12期1174-1180,共7页Journal of Chinese Integrative Medicine

摘　　要：目的:观察黄芪注射液、黄芪甲苷、芒柄花素对 basal-like 乳腺癌细胞株 MDA-MB-468和 MDA-MB-231增殖和磷酸化 Akt(phospho-Akt(?)p-Akt)蛋白表达的影响。方法:用不同浓度的黄芪注射液、黄芪甲苷和芒柄花素对乳腺癌细胞株 MDA-MB-468和 MDA-MB-231进行干预,以不加药物干预的细胞作为空白对照。用甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)法观察药物对细胞增殖的怍用。再选取药物抑制细胞增殖的最佳浓度,用细胞内蛋白质印迹法观察对 p—Akt(Thr308)和 p-Akt(Ser473)蛋白表达水平的影响。结果:根据 MTT 比色法实验结果确定黄芪注射液、黄芪甲苷、芒柄花素和黄芪甲苷配伍芒柄花素的最佳浓度,黄芪注射液终浓度为生药含量1 g/mL,黄芪甲苷终浓度为80 μg/mL,芒柄花素终浓度为40 μg/mL,黄芪甲苷:芒柄花素为1:4.干预 MDA-MB-468细胞1d 后,各用药组 p-Akt(Thr 308)和 p-Akt(Ser 473)蛋白表达水平下调,与阴性对照组比较,差异有统计学意义(P<0.05);药物干预2 d 后,各用药组 p—Akt(Thr 308)水平显著下调(P<0.05),其中以芒柄花素组、黄芪甲苷+芒柄花素组下调最为明显;芒柄花素和黄芪甲苷+芒柄花素明显下调 p-Akt(Ser 473)蛋白表达。干预 MDA-MB-231细胞1 d 后,与阴性对照组比较,芒柄花素组和黄芪甲苷+芒柄花素组 p-Akt(Thr 308)蛋白水平有明显的下调;在药物干预2 d 后,黄芪注射液、芒柄花素和黄芪甲苷+芒柄花素组 p-Akt(Thr 308)蛋白水平都有明显下调。各用药组 p-Akt(Ser 473)蛋白水平与阴性对照组比较,差异无统计学意义。结论:黄芪注射液、黄芪甲苷、芒柄花素对乳腺癌细胞株 MDA-MB-468和 MDA-MB-231均有抑制增殖的作用,但抑制作用程度及其浓度存在差异,其抑制细胞增殖的机制可能与下调 Akt 的磷酸化有关。Objective： To observe the effects of Astragalus injection, astragaloside Ⅳ and formononetin on proliferation and Akt phosphorylation of basal-like human breast carcinoma cell lines MDA-MB-468 and MDA-MB-231. Methods： The effects of different concentrations of Astragalus injection, astragaloside Ⅳ and formononetin on proliferation of breast cancer cell lines were assayed by methyl thiazolyl tetrazolium （MTT） assay, and their effects on phospho-Akt were assayed by in-cell Western blot method. Results： The results of the MTT assay showed that the best concentrations of Astragalus injection. astragaloside Ⅳ formononetin and astragaloside Ⅳ plus formononetin were 1 g/mL, 80 μg/mL, 40 μg/mL and 10 μg/mL plus 40 μg/mL respectively. After 1- or 2-day culture, Astragalus injection, astragaloside Ⅳ,formononetin and astragaloside IV plus formononetin decreased the expressions of p-Akt （Thr 308） and p-Akt （Ser 473） in MDA-MB-468 ceils. Formononetin and astragaloside IV plus formononetin down-regulated the expression of p-Akt （Thr 308） protein in MDA-MB-231 cells after 1- and 2-day culture, but had no effects on the expression of p-Akt （Ser 473） protein in MDA-MB-231 cells. Conclusion： Astragalus injection, astragaloside Ⅳ and formononetin can inhibit proliferation of breast cancer cell lines MDA-MB-468 and MDA-MB-231, and the antiproliferation effects vary according to their concentrations. And the antiproliferation mechanisms may be related to their down-regulation effects on Akt phosphorylation.

关 键 词：黄芪注射液 黄芪甲苷 芒柄花素 乳腺癌 细胞增殖 磷酸化AKT 

分 类 号：R285[医药卫生—中药学] R737.9[医药卫生—中医学]