摘要
目的探讨雷公藤内酯醇(triptolide,TP)诱导皮肤T细胞淋巴瘤蕈样肉芽肿细胞株Hut102的凋亡及其机制。方法体外培养的Hut102细胞与12.5、25、50、100nmol/L雷公藤内酯醇共同孵育,分别在24、48、72h用MTT法检测Hut102细胞的增殖抑制率,AnnexinV-FITC/PI双染流式细胞术分析细胞凋亡,Westernblot检测Hut102细胞中磷酸化p38丝裂原活化蛋白激酶(p-p38)、caspase-3蛋白的表达水平。结果不同浓度雷公藤内酯醇(12.5、25、50、100nmol/L)作用Hut102细胞24h后的增殖抑制率分别为8.67%、30.02%、52.54%、59.62%。AnnexinV-FITC/PI双染流式细胞术检测经25、50、100nmol/L雷公藤内酯醇处理24h后Hut102细胞凋亡率分别为9.19%、21.49%、26.34%,当50nmol/L雷公藤内酯醇组预先用p38MAPK特异性抑制剂SB203580干预后,Hut102细胞凋亡率下降为14.35%。不同浓度雷公藤内酯醇(50、100nmol/L)处理Hut102细胞后p38、caspase-3蛋白的表达被激活,同样50nmol/L雷公藤内酯醇组也预先用p38MAPK特异性抑制剂SB203580干预,结果p-p38的表达降低,caspase-3激活被抑制。结论雷公藤内酯醇可以抑制Hut102细胞的增殖,其作用机制可能是通过激活p38MAPK信号通路使部分Hut102细胞产生凋亡。
Objective To study the effect of triptolide on apoptosis of human cutaneous T-cell lymphoma Hut102 cells and its mechanism.Methods Cutaneous T-cell lymphoma cell line Hut102 was cocultured with triptolide solution at the terminal concentration of 12.5,25,50 and 100 nmol/L,respectively,in 96-well plates.At 24,48,and 72 h after co-culture,inhibitory effect of triptolide on Hut102 cells was detected by MTT assay,apoptosis rate of Hut102 cells was assessed by Annexin V-FITC/PI,and expression of phosphorylated p38 mitogen-activated protein kinase（p-p38MAPK） and caspase-3 was determined by Western blot analysis.Results Triptolide inhibited the proliferation of Hut102 cells in a concentration-and time-dependent manner.Annexin V-FITC/PI showed that the apoptotic rate of Hut102 cells was 9.19%,21.49% and 26.34%,respectively,in 24 h after they were treated with triptolide at the concentration of 25,50 and 100 nmol/L,respectively.However,the apoptosis rate of Hut102 cells was decreased by 14.35% after they were interfered with the p38MAPK inhibitor（SB203580）.The expression of p-p38MAPK and caspase-3 was activated in Hut102 cells by different concentrations of triptolide（50 and 100 nmol/L）.However,the expression of phosphorylated p38MAPK was decreased and the activity of caspase-3 was inhibited by triptolide at the concentration of 50 nmol/L interfered with SB203580.Conclusion Triptolide can inhibit the proliferation of Hut102 cells by activating the p38MAPK signal pathway,thus inducing apoptosis of certain Hut102 cells.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第19期2109-2111,共3页
Journal of Third Military Medical University
作者简介
[通信作者]郝进，电话：（023）63693411，E-mail：haojin_cq@163．com
