CAJ | 학술논문

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West-ern 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。
목적：연구적설초산대 T98G 세포조망급자서적영향。방법 MTT법분석적설초산대T98G세포증식적영향；Hoechst 33258염색관찰세포형태；류식세포술검측세포조망화세포자서；Western blot검측조망화자서통로상관인자적표체。결과적설초산대 T98G 세포증식유억제작용, IC50치위46．3μmol · L-1。 Annexin-V/7-AAD 쌍염화West-ern blot결과현시,적설초산가인기Akt표체하조、선립체막전위하강화Caspase-3격활,진이유도T98G세포조망。동시,AA가억제T98G세포자서,표현위강저MDC형광강도,억제Beclin-1、LC3-Ⅱ、Atg표체。통과록규상조Beclin-1화LC3-Ⅱ적표체,적설초산억제자서작용피조단,동시촉조망작용야명현강저。결론적설초산가동시유도T98G세포조망화억제자서,병차억제자서작용능구촉진세포조망,진이억제T98G세포증식。
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.