雷公藤内酯醇协同地塞米松诱导蕈样肉芽肿hut-102细胞凋亡及其机制

Apoptosis of mycosis fungoides hut-102 cells induced by triptolide in combination with dexamethasone and its mechanism

目的:探讨雷公藤内酯醇(Triptolide,TP)与地塞米松联合对蕈样肉芽肿(Mycosis fungoides,MF)hut-102细胞体外增殖、凋亡是否具有协同作用及其机制。方法:MTT法检测细胞存活率;流式细胞术检测细胞凋亡率;蛋白印迹法检测糖皮质激素受体(Glucocorticoid receptor,GR)和凋亡相关蛋白Caspase-3表达。结果:①不同浓度TP(6.25、12.5、25nmol/L)和地塞米松(10-7、10-6、10-5mol/L)对hut-102细胞均有增殖抑制作用,与时间、浓度相关;12.5nmol/LTP与不同浓度地塞米松联合,细胞抑制率随地塞米松浓度升高呈现协同作用。②TP和地塞米松均能诱导hut-102细胞凋亡,联合用药凋亡率明显高于单药,具有交互效应。③10-5mol/L地塞米松作用hut-102细胞后,GR表达降低(P<0.05),联合用药(12.5nmol/LTP+10-5mol/L地塞米松)GR表达上调、Caspase-3激活增强(P<0.05)。结论:TP联合地塞米松可明显抑制hut-102细胞增殖,增强诱导凋亡,具有协同效应,其作用机制可能与GR表达上调、Caspase-3激活增强有关。

Objective：To investigate the effect of triptolide combined with dexamethasone on proliferation and apoptosis of the mycosis fungoides hut-102 cells and the possible mechanisms.Methods：The viability of hut-102 cells was measured by MTT assay;the apoptotic rate of hut-102 cells was detected by flow cytometry.The expression levels of Glucocorticoid receptor（GR）and apoptosis-related protein Caspase-3 were determined by Western blot analysis.Results：①MTT assays showed that different concentrations of triptolide or dexame thasone can effectively inhibit the proliferation of hut-102 cells in a dose and time-dependent manner.When triptolide（12.5 nmol/L）was co-administered with dexamethasone,the inhibition rate increased along with the concentration of dexamethasone.And we observed positive cooperative inhibitory effects of triptolide and dexamethasone on the proliferation of hut-102 cells.②Both triptolide and dexame thasone can independently promote apoptosis of the hut-102 cell.And the apoptotic rate in the combination group was higher than single group（P0.01）.③Compared with the significantly decreased expression of Glucocorticoid receptor（GR）in hut-102 cells in dexameth asone group,the expression of GR and activated caspase-3 increased in the combination group（P0.05）.Conclusion：The apop tosis of hut-102 cells induced by dexamethasone was enhanced by triptolide,which maybe connected with the increased expression of GR and activated Caspase-3 in hut-102 cells.