积雪草酸通过改变自噬作用促进胶质瘤细胞T98G凋亡的实验研究  

Asiatic acid induces apoptosis in T98G human glioblastoma cells by changing autophagy

作　　者：李欢[1] 管福琴[1] 陈雨[1] 印敏[1] 孙浩[1] 王鸣[1] 冯煦[1] 单宇[1] 

机构地区：[1]江苏省中国科学院植物研究所南京中山植物园江苏省农业种质资源保护与利用平台,江苏南京210014

出　　处：《中国药理学通报》2015年第10期1363-1368,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81402829);江苏省自然科学基金资助项目(No BK20131338);江苏省盐土生物资源研究重点实验室开放课题(No JKLBS2013003);江苏省药用植物研究开发中心开放基金(No药201201)

摘　　要：目的研究积雪草酸对T98G细胞凋亡及自噬的影响。方法 MTT法分析积雪草酸对T98G细胞增殖的影响;Hoechst 33258染色观察细胞形态;流式细胞术检测细胞凋亡和细胞自噬;Western blot检测凋亡和自噬通路相关因子的表达。结果积雪草酸对T98G细胞增殖有抑制作用,IC50值为46.3μmol·L-1。Annexin-V/7-AAD双染和Western blot结果显示,积雪草酸可引起Akt表达下调、线粒体膜电位下降和Caspase-3激活,进而诱导T98G细胞凋亡。同时,AA可抑制T98G细胞自噬,表现为降低MDC荧光强度,抑制Beclin-1、LC3-Ⅱ、Atg表达。通过氯喹上调Beclin-1和LC3-Ⅱ的表达,积雪草酸抑制自噬作用被阻断,同时促凋亡作用也明显降低。结论积雪草酸可同时诱导T98G细胞凋亡和抑制自噬,并且抑制自噬作用能够促进细胞凋亡,进而抑制T98G细胞增殖。Aim To investigate the effect of asiatic acid on apoptosis and autophagy in human glioblastoma T98G cells. Methods MTT colorimetry was employed to assay the cellular proliferating activity. The fluores-cence microscope and Hoechst 33258 staining were used to detect the morphological changes. The cell ap-optosis and autophagy were analyzed by flow cytometry with Annexin-V/7-AAD and MDC staining respective-ly. The expressions of associated proteins were detected by Western blot to analyze the mechanism of apoptosis and autophagy. Results MTT assay showed that the growth of T 9 8 G cells was inhibited by asiatic acid ( IC50 =46. 3 μmol · L-1 ) . Annexin V/7-AAD stai-ning and Western blot revealed that asiatic acid in-duced apoptosis in T98 G cells by reducing the expres-sion of Akt, decreasing the mitochondrial membrane potential, and increasing the expression of Caspase-3. MDC staining and Western blot showed that the per-centage of MDC-positive cells was decreased and the expressions of Beclin-1 , LC3-II and Atgs were inhibi-ted by asiatic acid treatment. 5 μmol·L-1 chloroquine was used to up-regulate the expressions of LC3-Ⅱand Beclin-1 . Asiatic acid-inhibited autophagy was blocked and the total apoptotic rate was reduced remarkably. Conclusion Asiatic acid suppresses T98 G cells pro-liferation by inducing apoptosis and inhibiting cell au-tophagy, and the very role of inhibiting autophagy could promote apoptosis to a certain extent.

关 键 词：积雪草酸 T98G细胞 凋亡 自噬 机制 相互作用 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]