芦荟大黄素对Jurkat T细胞增殖和凋亡的作用及机制

目的 研究芦荟大黄素对Jurkat T细胞的增殖抑制和凋亡诱导作用,并探讨可能的机制.方法 应用细胞计数测定增殖,Hoechst/PI双染法及DNA片断化分析细胞凋亡特征,流式细胞术检测细胞周期分布.进一步应用二氢乙锭荧光染色测量活性氧产物,Western blotting检测胞浆细胞色素C(Cyt-c)的量,比色法检测caspase-3和caspase-9活性.结果 芦荟大黄素剂量和时间依赖性地抑制Jurkat T细胞增殖.芦荟大黄素诱发细胞核皱缩,核DNA片断化,细胞周期出现亚G1期凋亡峰,且停滞于G2/M期.芦荟大黄素介导细胞内活性氧水平显著提高,线粒体释放Cyt-c量显著增加,caspase-3和caspase-9活性显著增强.结论 芦荟大黄素剂量依赖性地抑制Jurkat T细胞增殖并诱导其凋亡.诱导凋亡的机制中,包括增加活性氧产物和线粒体损伤途径.

Effects and mechanism of aloe-emodin on proliferation and apoptosis of Jurkat T cell line

Objective To explore the effects and mechanism of aloe-emodin on proliferation and apoptosis of human leukemia cell line Jurkat T.Methods The effect of aloe-emodin on the proliferation of Jur-kat T cells was studied through trypan blue exclusion.The effect of aloe-emodin on cell apoptosis was observed by Hoechst/PI dyeing and DNA fragmentation analysis.The distribution of cell cycle was examined by flow cytometry(FCM).ROS Production was analyzed using dihydroethiudium fluorescent staining.Cytosolic C(Cyt-c) was measured by Western blotting and cytosolic caspases-3 and caspase-9 activities were estimated through colorimetric assay.Results Aloe-emodin inhibited Jurkat T cells proliferation on a time-and dose-dependent manner.Aloe-emodin induced nucleus crimple of some cells and DNA fragmentation.Apoptosis peak sub G1 appeared and the cell cycle was arrested in G₂/M after aloe-emodin treatment.Aloe-emodin mediated intracellular ROS level elevating,Cyt-c release from mitochondrion enhancing and cytosolic caspase-3 and caspase-9 activity increasing.Conclusion Aloe-emodin inhibits Jurkart T cell line proliferation and induces cell apoptosis on a dose-dependent manner.The mechanism of aloe-emodin induced cell apoptosis involves increasing ROS production and mitochondrial impairment pathway.