Effects of miR-199a in regulation of Sirt1 on oxidative damage of SH-SY5Y nerve cells
Objective To investigate the effects of miR-199a in regulation of Sirt1 on oxidative damage of SH-SY5Y nerve cells.Methods Cultured SH-SY5Y nerve cells were stimulated with H2O2(1 200 μmol/L)for 3 h,6 h,9 h,12 h and 15 h respectively.Cell viability was detected by MTT assay and intracellular ROS levels were de-tected by DCF-DA probe.SH-SY5Y nerve cells were transfected with miR-199a mimics and inhibitors for 48 h,the expression of Sirt1 protein was detected by Western blot.SH-SY5Y nerve cells were transfected with miR-199a mimics and inhibitors for 48 h respectively and stimulated with H2O2(1 200 μmol/L)for 12 h.The activity of su-peroxide dismutase(SOD)and the content of malondialdehyde(MDA)were detected by enzyme biochemical meth-od.Result When SH-SY5Y nerve cells were stimulated by H2O2 at different time,the cell viability and intracellular ROS content increased in a time-dependent manner.The miR-199a mimicon inhibited Sirt1 protein expression in SH-SY5Y neurons,the miR-199a inhibitor increased Sirt1 protein expression in SH-SY5Y neurons.When SH-SY5Y nerve cells were in a state of oxidative damage,miR-199a mimics decreased SOD content and increased MDA content,while miR-199a inhibitor increased cell vitality,decreased intracellular ROS content,increased SOD content and decreased MDA content.Conclusion When SH-SY5Y nerve cells are in the state of oxidative damage,the regulation of Sirt1 by miR-199a further aggravated the oxidative damage of SH-SY5Y nerve cells.
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