[关键词]
[摘要]
目的 探讨白头翁总皂苷碱水解产物(PAHS)的抗胶质瘤作用及机制。方法 采用MTT法、集落形成实验分别检测细胞存活率和集落形成率,同时采用Giemsa染色观察PAHS对肿瘤细胞形态学变化;采用hoechest33342染色观察人脑胶质母细胞瘤细胞株U251凋亡细胞形态变化;用流式细胞仪检测细胞的凋亡;用Western Blot法检测Bcl-2和Caspase-3蛋白的表达变化。结果 PAHS可以剂量依赖性的抑制U251细胞的增殖,抑制U251细胞集落的形成;PAHS(6.25,9.375,12.5 μg·mL-1)可以诱导U251细胞凋亡,抑制率分别为(6.00±2.05) %、(46.19±0.24) %、(78.26±2.10) %。PAHS可以上调Caspase-3、Bcl-2等蛋白的表达。结论 PAHS具有良好的抗胶质瘤作用,其机制可能与调节Bcl-2和Caspase-3蛋白的表达有关。
[Key word]
[Abstract]
To investigate antitumor effect and mechanism of alkali hydrolysate of total saponins from Pulsatilla chinensis on human glioma U251 cells. Methods MTT assay and colony formation test were used to examine the growth of human glioma cells and colony formation rate. Meanwhile,Giemsa staining was used to observe the morphology of tumor cells,flow cytometry was used to detect the apoptosis of tumor cells,Hoechst 33342 assay kits were adopted to observe the morphology of apoptotic cells,and the expression levels of Caspase-3 and Bcl-2 protein were evaluated by Western blot assay. Results Alkali hydrolysate of total saponins from Pulsatilla chinensis exhibited an inhibition effect on the proliferation and colony formation of U251 cells in dose-dependent manner. The alkali hydrolysate at the dosage of 6.25,9.375,12.5 μg·mL-1 could induce apoptosis of U251 cells,and the apoptotic rates were(6.00±2.05)%,(46.19±0.24)% and(78.26±2.10)%,respectively. Western blotting results showed that the alkali hydrolysate could up-regulate the expression levels of Caspase-3 and Bcl-2. Conclusions Alkali hydrolysate of total saponins from Pulsatilla chinensis exerts satisfactory anti-glioma action,and the mechanism might be related with the regulation of Bcl-2 and Caspase-3 protein expression.
[中图分类号]
R285.5
[基金项目]
国家自然科学基金项目(81273402);重大新药创制国家科技重大专项(2011ZX11102-001-19);江苏省自然科学基金(BK2012620);2013年苏州大学大学生创新创业训练计划(2013xj060)。