Expression and significance of MeCP2 and TH protein in normal SH-SY5Y cells and Parkinson's disease cell model / 国际检验医学杂志

ABSTRACT

Objective To investigate the expression and significance of TH and MeCP2 protein in normal SH-SY5Y cells and Parkinson's disease cell model.Methods The experiment was divided into 3 groupsblank control group,non transfection;model control group;pEGFP-N1-MeCP2 group,transfection solution added to the recombinant pEGFP-N1-MeCP2 plasmid.In addition to the blank group,the remaining two groups were added 50.0 μmol/L 6-OHDA treatment for 24 hours.CCK-8 assay and flow cytometry were used to detect cell activity and apoptosis in normal SH-SY5Y cells and the SH-SY5Y cell with up-regulation of MeCP2 induced by 6-OHDA.Immunofluorescence and Western blot analysis were used to detect the changes of MeCP2 protein and TH protein expression in each group of SH-SY5Y cells.Results As for the cell survival and apoptosis of SH-SY5Y cells detected by CCK-8 assay and induced 6-OHDA of flow cytometry and SH-SY5Y cells with up-regulation of MeCP2 in model control group,compared to those in pEGFP-N1-MeCP2 group and the thank control group,the difference was statistically significant (P＜0.05).The expression of MeCP2 and TH in SH-SY5Y induced 6-OHDA cells was detected by double immunofluorescence,it was found that in model control group,with 6-OHDA (50.0 μmol/L) inducing for 24 hours,and the expression of MeCP2 and TH decreased significantly (P＜0.05).After Western blot was used to detect the up-regulation of MeCP2 expression,in 6-OHDA induced SH-SY5Y cells of MeCP2 and TH expression,the expression of MeCP2 and TH protein in pEGFP-N1-MeCP2 group and blank control group was statistically different from that in model control group (P＜0.01).Conclusion The transfection of pEGFP-N1-MeCP2 recombinant plasmid could inhibit the apoptosis of SH-SY5Y cells induced by 6-OHDA,improve the expression of TH and increase the cell survival rate.