title: [Oxidative stress and autophagy in SK-N-SH cells induced by manganese chloride or 1-methyl-4-phenylpyridinium: a comparative analysis].
abstract: Objective:To investigate the effect of manganese chloride (MnCl(2)) or 1-methyl-4-phenylpyridinium (MPP (+)) on oxidative stress and autophagy in human neuroblastomaSK-N-SH cells and the mechanism of the neurotoxicity of manganese.Methods:SK-N-SH cells were treated with MnCl(2) or MPP(+) at doses of 0.062 5, 0.125, 0.25, 0.5, 1.0, and 2.0 mmol/L for 24 hours, and MTT assay was used to measure cell viability. The cells weretreated with MnCl(2) or MPP(+) at doses of 0.125, 0.25, and 0.5 mmol/L for 24 hours, and flow cytometry was used to measure the content of reactive oxygen species (ROS) in cells, a laser scanning confocal microscope was used to observe autophagosome in cells, and Western blot was used to measure the expression of autophagy-related proteins P62 and LC3-II/LC3-I.Results:Compared with the control group, the 0.0625-2.0 mmol/L MnCl(2) and 0.125-2.0 mmol/L MPP (+) treatment groups had significant reductions in the viability of SK-N-SH cells, and the 0.25-2.0 mmol/L MnCl(2) treatment groups had significantly lower viability than the groups treated with the same doses of MPP(+) (allP<0.05) . Compared with the control group, the 0.125-0.25 mmol/L MnCl(2) and 0.125-0.5 mmol/L MPP(+) treatment groups had significant increases in the content of ROS, and the 0.25-0.5 mmol/L MPP(+) treatment groups had significantly higher content of ROS than the groups treated with the same doses of MnCl(2) (allP<0.05) . Compared with the control group, the 0.25-0.5 mmol/L MnCl(2) andMPP(+) treatment groups had significant increases in autophagy-related proteins LC3-II/LC3-I and significant reductions in P62 expression; the 0.125-0.5 mmol/L MPP(+) treatment groups had significantly higher LC3-II/LC3-I than the groups treated with the same doses of MnCl(2), and the 0.125 and 0.25 mmol/L MPP (+) treatment groups had significantly lower P62 expression than the groups treated with the same doses of MnCl(2) (allP<0.05) .Conclusion:Both MnCl(2) and MPP(+) can induce oxidative stress and autophagy in SK-N-SH cells, and MPP(+) has a significantly greater inductive effect on autophagy of SK-N-SH cells than MnCl(2).
authors.affiliation: School of Public Health, Peking University, Beijing 100191, China.
authors.forename: W L
authors.initials: WL
authors.lastname: Liu
authors.forename: C S
authors.initials: CS
authors.lastname: Dou
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.forename: P
authors.initials: P
authors.lastname: Zhao
authors.forename: J L
authors.initials: JL
authors.lastname: Fu
authors.forename: B Y
authors.initials: BY
authors.lastname: Yao
authors.forename: Z C
authors.initials: ZC
authors.lastname: Zhou
mesh_terms: D015655:1-Methyl-4-phenylpyridinium / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy / Q000187:drug effects*; D045744:Cell Line, Tumor / Q000187:drug effects*; D002712:Chlorides / Q000633:toxicity*; D006801:Humans; D017895:Manganese Compounds; D009447:Neuroblastoma; D018384:Oxidative Stress / Q000187:drug effects* / Q000502:physiology
chemical_list: D002712:Chlorides; D017895:Manganese Compounds; C025340:manganese chloride; D015655:1-Methyl-4-phenylpyridinium
keywords: Autophagy; MPP(+); Manganese; Oxidative stress; SK-N-SH cells