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目的研究不同价态锰化合物对神经母细胞瘤细胞(SH-SY5Y)DNA损伤作用、细胞凋亡和细胞周期的影响,以及抗氧化剂硒的保护作用,从而探讨锰的神经毒作用机制。方法将培养的SH-SY5Y细胞随机分为染毒的二价锰和三价锰化合物(0.5 mmol/L)组、空白对照组、硒干预组。培养24 h后应用单细胞凝胶电泳检测细胞DNA链断裂情况。48 h后用流式细胞仪测定细胞凋亡率、细胞周期分布。结果三价锰组DNA损伤程度比二价锰组严重。二价锰和三价锰组均出现亚二倍体的凋亡峰;细胞周期分布也发生改变,与对照组相比S期细胞明显增多,差异有统计学意义(P<0.01);三价锰比二价锰更易引起细胞凋亡(P<0.01)和细胞周期变化(P<0.05)。硒可降低锰化合物引起的细胞DNA损伤、凋亡率和细胞周期中S期细胞数。结论不同价态锰对神经细胞均有损伤作用,且三价锰大于二价锰。硒对锰致细胞DNA损伤、凋亡和细胞周期改变有一定的保护作用,提示氧化应激可能参与了锰的毒作用。
Objective To investigate the effect of different valence manganese compounds on the DNA damage, apoptosis and cell cycle of neuroblastoma cells (SH-SY5Y) and the protective effect of antioxidants on selenium, so as to explore the neurotoxic mechanism of manganese. Methods The cultured SH-SY5Y cells were randomly divided into two groups: manganous and manganese (0.5 mmol / L), blank control group and selenium intervention group. After cultured for 24 h, single cell gel electrophoresis was used to detect DNA strand breaks. After 48 h, the apoptosis rate and the cell cycle distribution were determined by flow cytometry. Results The DNA damage in the trivalent manganese group was more serious than that in the divalent manganese group. The apoptotic peak of subdiploid appeared in both manganese (Mn) and manganese (Mn) groups. The cell cycle distribution also changed. Compared with the control group, S phase cells increased significantly (P <0.01) Manganese more easily induced apoptosis (P <0.01) and cell cycle changes than Mn (P <0.05). Selenium can reduce the DNA damage caused by manganese compounds, apoptosis rate and cell cycle in S phase cells. Conclusion Different valences of manganese have a damaging effect on nerve cells, and the content of trivalent manganese is larger than that of bivalent manganese. Selenium has certain protective effect on DNA damage, apoptosis and cell cycle of manganese induced by manganese, suggesting that oxidative stress may be involved in the toxic effect of manganese.