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目的探讨茶多酚中主要活性成分没食子儿茶素没食子酸酯(EGCG)对百草枯诱导人神经母细胞瘤细胞株SK-N-SH细胞凋亡的保护作用。方法培养的SK-N-SH细胞给予400μmol·L-1百草枯诱导细胞凋亡。实验分为6组:空白对照组、百草枯模型组、维生素E(10μmol·L-1)组和3个EGCG(1、5、10μmol·L-1)剂量组。以药物处理细胞2h后,加入百草枯,72h后MTT法检测细胞活力,取培养细胞上清液测定乳酸脱氢酶(LDH)漏出量,Hoechst 33258荧光染色法和流式细胞术检测细胞凋亡情况。结果与空白对照组相比,百草枯明显降低细胞活力(P<0.01),增加LDH漏出量(P<0.01),细胞膜结构不完整,出现空泡等凋亡现象,细胞凋亡发生率达到30.5%。EGCG处理后,显著提高细胞活力,减少LDH的漏出和降低细胞凋亡率(P<0.01),其中10μmol·L-1组的作用明显高于5μmol·L-1组或1μmol·L-1组(P<0.05或P<0.01)。结论EGCG具有抑制百草枯诱导的SK-N-SH细胞凋亡作用。
Objective To investigate the protective effect of EGCG, a major active ingredient in tea polyphenols, on paraquat-induced apoptosis of human neuroblastoma SK-N-SH cells. Methods SK-N-SH cells were treated with 400 μmol·L -1 paraquat to induce apoptosis. The experiment was divided into 6 groups: blank control group, paraquat model group, vitamin E (10μmol·L-1) group and 3 EGCG (1,5,10μmol·L-1) dose groups. After treated with drug for 2 hours, paraquat was added and cell viability was measured by MTT method 72 hours later. The amount of lactate dehydrogenase (LDH) leakage was measured by culture supernatant. Hoechst 33258 fluorescence staining and flow cytometry were used to detect apoptosis Happening. Results Paraquat significantly decreased cell viability (P <0.01), increased LDH leakage (P <0.01), cell membrane structure was incomplete, vacuole and other apoptotic phenomena were observed, and the incidence of apoptosis reached 30.5 %. EGCG treatment significantly increased cell viability, decreased leakage of LDH and decreased apoptosis rate (P <0.01). The effect of 10μmol·L-1 group was significantly higher than that of 5μmol·L-1 group or 1μmol·L-1 group (P <0.05 or P <0.01). Conclusion EGCG can inhibit paraquat-induced apoptosis of SK-N-SH cells.