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目的研究不同剂量的锰(Mn)对人神经母细胞瘤细胞SH-SY5Y细胞生长增殖的影响以及诱导自噬的情况。方法体外培养SH-SY5Y细胞,以200～1 000μmol/L MnCl2处理后进行四唑盐比色实验(MTT)观察MnCl2对细胞的抑制作用,MDC染色法检测自噬小泡形成情况,Western blot检测自噬标志蛋白LC3和Bclin-1的表达和活化情况。DCFH-DA染色检测活性氧(ROS)含量。结果 MTT结果显示200～1 000μmol/L MnCl2对细胞的生长增殖有显著的抑制作用。Western blot结果显示MnCl2处理后LC3蛋白表达增加并且活化,Bcelin-1蛋白表达增加,细胞产生自噬小泡。与对照组相比,200、400和800μmol/L MnCl2组ROS含量明显升高。结论一定剂量的MnCl2对SH-SHY5Y细胞的生长增殖有明显的抑制作用,这一作用可能是通过破坏胞内氧化还原平衡导致ROS大量产生继而诱发细胞发生自噬性的死亡。
Objective To study the effect of different doses of manganese (Mn) on the growth and proliferation of human neuroblastoma SH-SY5Y cells and the induction of autophagy. Methods SH-SY5Y cells were cultured in vitro and treated with 200 ~ 1 000μmol / L MnCl2 for MTT assay. The inhibitory effect of MnCl2 on cells was detected by MTT assay. The formation of autophagic vesicles was detected by MDC staining. Expression and activation of autophagy marker proteins LC3 and Bclin-1. DCFH-DA staining was used to detect the content of reactive oxygen species (ROS). Results MTT results showed that 200 ~ 1 000μmol / L MnCl2 significantly inhibited cell growth and proliferation. Western blot results showed that the expression of LC3 protein was increased and activated after MnCl2 treatment, and the expression of Bcelin-1 protein was increased. Autophagy vesicles were produced by the cells. Compared with the control group, ROS levels in 200, 400 and 800μmol / L MnCl2 groups were significantly increased. Conclusion A certain dose of MnCl2 significantly inhibits the growth and proliferation of SH-SHY5Y cells. This effect may be due to the destruction of intracellular redox balance leading to the massive production of ROS, which in turn leads to autophagic cell death.