双氢青蒿素对UVB诱导的人皮肤HaCaT细胞凋亡的保护作用

目的:明确双氢青蒿素(DHA)对UVB诱导人皮肤HaCaT细胞凋亡的保护作用。方法:将体外培养的HaCaT细胞分为空白对照组,UVB照射组和UVB+DHA(20、40、60、80μmol/L)组,采用MTT法检测细胞系的增殖情况,流式细胞仪检测细胞的凋亡率,RT-PCR法检测抗凋亡基因survivin和促凋亡基因caspease-3(激活型)mRNA水平,Western检测相关蛋白表达水平。结果:30 mJ/cm~2 UVB照射24 h后,UVB组和UVB+DHA(20、40、60、80μmol/L)组细胞的增殖率分别为空白对照组的(50.13±2.42)%,(60.23±2.30)%,(69.24±3.19)%,(79.37±2.47)%和(70.41±2.24)%。UVB组和UVB+60μmol/L DHA组中HaCaT细胞凋亡率分别为(46.7±3.2)%和(23.71±1.2)%。与UVB组比较,UVB+60μmol/L DHA组中HaCaT细胞caspase-3 mRNA及蛋白表达降低、survivin mRNA及蛋白表达升高。结论:DHA可抑制UVB所致HaCaT细胞的凋亡,其机制可能与survivin和caspase-3有关。

Protective effect of dihydroartemisinin on HaCaT cell apoptosis induced by UVB

Objective:To determine the protective effect of dihydroartemisinin ( DHA) on HaCaT cells ap-optosis induced by UVB. Methods:HaCaT cells cultured in vitro were divided into blank control group, UVB group and UVB+DHA (20, 40, 60, 80 μmol/L) group. The proliferation of HaCaT cells, cell apoptosis, mRNA levels of anti-apoptotic gene survivin and pro-apoptotic gene caspease-3 and corresponding protein levels were detected by MTT, flow cytometric analysis, qRT-PCR and Western blotting. Results: After the HaCaT cells was irradiated for 24 hs, the proliferation of the cells in the UVB group and UVB+DHA ( 20, 40, 60, 80 μmol/L) groups was (50.13±2.42)%, (60.23±2.30)%, (69.24±3.19)%, (79.37±2.47)% and (70.41±2.24)% of the blank control group. The apoptosis rate in the UVB group and UVB+60μmol/L DHA group was (46.7±3.2)% and (23.71±1.2)%. Compared with the UVB group, the mRNA and protein levels of pro-apoptotic gene caspease-3 in the UVB+60 μmol/L DHA group was decreased and the levels of anti-apoptotic gene survivin was increased. Conclusion: DHA inhibits the apoptosis of HaCaT cells induced by UVB, which may be related with caspase-3 and survivin.