Increase the expression of methyl cytosine binding protein-2 in SH-SY5Y cell line suppression apoptosis which after 6-hydroxydopamine-induced

Abstract

Objective Observed apoptosis in SH-SYSY cells and protein expression of tyrosine hydroxylase and methyl cytosine binding protein-2 (MeCP2),and explore the role of MeCP2 in parkinson' s disease.Methods In this study,to get the up-regulation of MeCP2 in SH-SY5Y cell lines,the eukaryotic expression vector construction,the cells transfected technology was used.Recombinant plasmid transfected to SH-SY5Y cells,analysis of the MeCP2 protein and endogenous tyrosine hydroxylase (TH) protein expression by Immunofluorescence and Western blotting.Detected cell activity and apoptosis in normal SH-SY5Y cells and the SH-SY5Y cell that up-regulation of MeCP2 which pretreated with 6-hydroxydopamine (6-OHDA) by cell counting kit-8 (CCK-8) assay,flow cytometrv.Detected the expression of MeCP2 protein and TH protein in each group of SH-SY5Y cells by Immunofluorescence and Western blotting analysis.Results The plasmid of pEGFP-N1-MeCP2 target the MeCP2 was transfected with SH-SY5Y cells to raise the expression of MeCP2,and then treated with 6-OHDA,the expression of MeCP2 and TH was not decline by the detection of immunofluorescence and Western blotting,the ratio of MeCP2/β-actin and TH/β-actin were 0.8865 ± 0.0486 and 0.4639 ± 0.0266 respectively.The result of flow cytometry showed that apoptosis of SH-SY5Y cells was approximately (0.40 ±0.00) ％ and CCK-8 detection showed that cell survival rate was approximately (96.25 ± 5.28).Conclusion MeCP2 plays an important role in the pathogenesis of Parkinson' s disease.The present data indicate that MeCP2 upregulation can reduce 6-OHDAinduced apoptosis,and increase the protein levels of TH in SH-SY5Y cells,suggesting that MeCP2 may he a potential therapeutic target for the treatment of PD. Key words: Parkinson's disease; Methyl cytosine binding protein-2; Tyrosine hydroxylase; 6-hydroxydopamine