摘要
目的:探究不同浓度的黄芪甲苷(Astragaloside IV,As-IV)对肝癌细胞的作用及其作用机制。方法:采用不同浓度黄芪甲苷,按照不同作用时间处理肝癌细胞;采用MTT法检测不同浓度黄芪甲苷对肝癌细胞的毒性作用;运用放线菌素D(Actinomycin D,ActD)诱导细胞凋亡构建肝癌细胞凋亡模型,采用Hoechst33258染色法和流式细胞术验证肝癌细胞凋亡情况,采用流式细胞术检测一定浓度的黄芪甲苷处理放线菌素D诱导后的肝癌细胞凋亡率,采用western blot法验证黄芪甲苷对蛋白激酶α(Protein kinase Cα,PKC-α)蛋白和细胞外调节蛋白激酶(ExtracelluLar reguLated protein kinases,ERK)通路蛋白的影响。结果:浓度为40μmol/L的黄芪甲苷处理24h细胞活性为0μmol/L时的80%,不会显著影响肝癌细胞活性,且黄芪甲苷对肝癌细胞活性的影响呈剂量和时间依赖性。放线菌素D浓度为2μg/mL处理24h时肝癌细胞凋亡率为45%,可有效诱导肝癌细胞凋亡。40μmol/L黄芪甲苷处理24h可有效抑制2μg/mL放线菌素D诱导的肝癌细胞凋亡,细胞凋亡率为8%,显著低于2μg/mL放线菌素D处理组细胞凋亡率27%(P<0.05),并可有效抑制细胞膜PKC-α和p-ERK1/2表达量升高。结论:黄芪甲苷可通过抑制PKC-α活化后从细胞质向细胞膜迁移,有效抑制放线菌素D诱导的肝癌细胞凋亡。
Objective:To investigate the effect of Astragaloside IV on hepatoma carcinoma cells and its mechanism.Methods:Using MTT assay to detect cell viability after Astragaloside IV treatment at different concentration and with different time.Hoechst33258 assay and flow cytometry assay were used to detect apoptosis of hepatoma carcinoma cells after actinomycin D treatment.Western blot was used to detect the relative expression of PKC-αand ERK proteins after Astragaloside IV treatment.Results:After40μmol/L Astragaloside IV treatment for 24 h,cell viability is about 80%of normal hepatoma carcinoma cells.Cell apoptosis rate is45%after 2μg/mL Actinomycin D treatment for 24 h.40μmol/L Astragaloside IV can effectively decrease ActD-induced HepG2 cell apoptosis rate from 8%to 27%and lead the rise of relative expression of membrane-PKC-αand p-ERK1/2.Conclusion:Astragaloside IV can inhibit ActD-induced cell apoptosis via inhibiting the migration of PKC from cytoplasm to cell membrane.
作者
刘松格
谷见法
潘琼
Liu Songge;Gu Jianfa;Pan Qiong(Department of Oncology,Zhengzhou Central Hospital,Zhengzhou 467000,China)
出处
《亚太传统医药》
2021年第2期27-30,共4页
Asia-Pacific Traditional Medicine
