Effect of miR-185 overexpression on MPP+-induced cell damage in Parkinson's disease model
Objective To investigate the effect of miR-185 overexpression on 1-methyl-4-phenylpyridine ion(MPP+)-induced cell damage in Parkinson's disease(PD)models and its mechanism.Methods PC12 cells were pas-saged in vitro.PC12 cells of the third generation in the logarithmic growth phase were randomly divided into the control group,the model group,NC mimics group,and miR-185 mimics group,respectively.Cells in the model group,the NC mimics group,and the miR-185 mimics group were induced with 1 mmol/L MPP+ to construct the PD cell models,while cells in the control group were not induced with MPP+.After 24 h of modeling,cells in the NC mimics group and miR-185 mimics group were transfected with NC mimics and miR-185 mimics,respectively.Cells from each group were collected.The expression of miR-185 was detected using RT-qPCR,the proliferation ability of cells cultured for 24,48,and 72 h was detected by CCK-8,the apoptosis rate was detected using flow cytometry,the intracellular glutathione peroxidase(GSH),superoxide dismutase(SOD),and malondialdehyde(MDA)were detected by ELISA,and the expression levels of Bax and Bcl-2 proteins were detected by Western blotting.Results Compared with the control group,the relative expression level of miR-185 and the proliferation activity of cells in the model group decreased after 24,48,and 72 h of culture,while the apoptosis rate increased(all P<0.05);the intracellular GSH and SOD content decreased,while the MDA content increased(both P<0.05);the relative expression level of Bcl-2 protein decreased,while the relative expres-sion level of Bax protein increased(both P<0.05).Compared with the NC mimics group,the relative expression level of miR-185 and cell proliferation activity after 24,48,and 72 h of culture increased,and apoptosis rate decreased in the miR-185 mimics group(all P<0.05);the intracellular GSH and SOD content increased,while the MDA content decreased(both P<0.05);the relative expression level of Bcl-2 protein increased,while the relative expression level of Bax protein decreased(both P<0.05).However,there were no statistically significant differences in the above indicators between the model group and the NC mimics group(all P>0.05).Conclusion Overexpression of miR-185 can promote the prolifera-tion and inhibit apoptosis of PD cells,and has a significant protective effect;alleviating oxidative stress damage,inhibit-ing Bax protein expression,and promoting Bcl-2 protein expression may be one of the mechanisms.