Cui Qiao, Tashiro Shin-ichi, Onodera Satoshi, Ikejima Takashi
China-Japan Research Institute of Medical Pharmaceutical Sciences, Shenyang Pharmaceutical University, Shenyang 110016, China.
Yao Xue Xue Bao. 2007 Jan;42(1):35-9.
To study the mechanism of downregulation of apoptosis by autophagy induced by oridonin in HeLa cells, the cell viability was measured by MTT method. DNA fragmentation was assayed by agarose gel electrophoresis. Autophagic and apoptotic ratio was determined by flowcytometric analysis. Protein expression was detected by Western blotting analysis. Oridonin induced both apoptosis and autophagy in HeLa cells. Apoptosis was upregulated by introduction of the inhibitor of autophagy, 3-methyladenine (3-MA). Addition of oridonin increased Bax/Bcl-2 expression ratio and cytochrome c, whereas the expression of SIRT-1 was decreased, and 3-MA pre-application enhanced these changes. Oridonin-induced autophagy antagonized apoptosis in HeLa cells through mitochondrial pathway.
为研究冬凌草甲素诱导HeLa细胞自噬从而下调细胞凋亡的机制,采用MTT法检测细胞活力。通过琼脂糖凝胶电泳检测DNA片段化。采用流式细胞术分析自噬和凋亡比例。通过蛋白质印迹分析检测蛋白质表达。冬凌草甲素可诱导HeLa细胞凋亡和自噬。自噬抑制剂3-甲基腺嘌呤(3-MA)的加入上调了细胞凋亡。加入冬凌草甲素可增加Bax/Bcl-2表达比例和细胞色素c,而沉默信息调节因子1(SIRT-1)的表达降低,预先应用3-MA可增强这些变化。冬凌草甲素诱导的自噬通过线粒体途径拮抗HeLa细胞凋亡。