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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 investigate the apoptosis of mycosis fungoides cell line Hut102 induced by triptolide (TP) and its mechanism. Methods Hut102 cells cultured in vitro were incubated with 12.5,25,50,100 nmol / L triptolide. The proliferation inhibition rate of Hut102 cells was detected by MTT assay at 24, 48 and 72 h respectively. The Annexin V-FITC / PI double staining flow cytometry Cell apoptosis was analyzed by Western blot. The expression of phospho-p38 mitogen-activated protein kinase (p-p38) and caspase-3 protein in Hut102 cells was detected by Western blot. Results The inhibitory rates of Hut102 cells treated with different concentrations of triptolide (12.5, 25, 50, 100 nmol / L) for 24 h were 8.67%, 30.02%, 52.54% and 59.62%, respectively. The apoptotic rates of Hut102 cells treated with 25, 50, 100 nmol / L triptolide for 24 h were 9.19%, 21.49%, 26.34% respectively by Annexin V-FITC / PI double staining flow cytometry. Alcohol group pre-p38MAPK specific inhibitor SB203580 intervention, Hut102 apoptosis rate decreased to 14.35%. The expression of p38 and caspase-3 protein in Hut102 cells treated with different concentrations of triptolide (50,100nmol / L) was also activated. Similarly, the triptolide group with 50nmol / L of triptolide was also pretreated with SB203580, a specific inhibitor of p38MAPK. The results showed that p The expression of -p38 is reduced and caspase-3 activation is inhibited. Conclusion Triptolide can inhibit the proliferation of Hut102 cells and its mechanism may be through the activation of p38 MAPK signaling pathway in some Hut102 cells.