MeCP2、TH蛋白在正常SH-SY5Y细胞和帕金森病细胞模型中的表达及其意义研究  

Expression and significance of MeCP2 and TH protein in normal SH-SY5Y cells and Parkinson′s disease cell model

作　　者：潘燕 

机构地区：[1]湖北省十堰市东风总医院神经内科3病区,湖北十堰442000

出　　处：《国际检验医学杂志》2018年第5期526-529,共4页International Journal of Laboratory Medicine

摘　　要：目的探讨MeCP2、TH蛋白在正常SH-SY5Y细胞和帕金森病细胞模型中的表达及其意义。方法实验分为3组:空白对照组,未转染;模型对照组;pEGFP-N1-MeCP2组,转染液加入重组pEGFP-N1-MeCP2质粒。除空白对照组外,其余二组均加入6-羟多巴胺(6-OHDA)50.0μmol/L处理24h。用CCK-8法、流式细胞仪检测正常SH-SY5Y细胞、上调MeCP2表达的SH-SY5Y细胞在6-OHDA诱导下细胞活性及细胞凋亡的变化;免疫荧光技术和Western blot法检测各组SH-SY5Y细胞中MeCP2蛋白、TH蛋白表达的变化。结果 CCK-8法检测及流式细胞仪6-OHDA诱导的SH-SY5Y细胞、上调MeCP2的SH-SY5Y细胞中细胞存活和凋亡情况发现,模型对照组与pEGFP-N1-MeCP2组、空白对照组相比,差异具有统计学意义(P<0.05)。双重免疫荧光检测6-OHDA诱导的SH-SY5Y细胞中的MeCP2、TH表达情况发现,模型对照组,6-OHDA(50.0μmol/L)诱导24h,MeCP2和TH的蛋白表达显著下降(P<0.05)。Western blot检测上调MeCP2表达后,6-OHDA诱导的SH-SY5Y细胞中的MeCP2、TH表达情况发现,pEGFP-N1-MeCP2组上调组和空白对照组中MeCP2、TH蛋白表达与模型对照组相比,差异具有统计学意义(P<0.01)。结论转染pEGFP-N1-MeCP2重组质粒可抑制6-OHDA诱导的SH-SY5Y细胞凋亡,提高TH的表达,提高细胞存活率。Objective To investigate the expression and significance of TH and MeCP2 protein in normal SH-SY5 Ycells and Parkinson′s disease cell model.Methods The experiment was divided into 3 groups：blank 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 50.0μmol/L 6-OHDA treatment for 24 hours.CCK-8 assay and flow cytometry were used to detect cell activity and apoptosis in normal SH-SY5 Ycells and the SH-SY5 Ycell 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-SY5 Ycells.Results As for the cell survival and apoptosis of SH-SY5 Ycells detected by CCK-8 assay and induced 6-OHDA of flow cytometry and SHSY5 Ycells 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-SY5 Yinduced 6-OHDA cells was detected by double immunofluorescence,it was found that in model control group,with 6-OHDA（50.0μmol/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-SY5 Ycells induced by 6-OHDA,improve the expression of TH and increase the cell survival rate.

关 键 词：帕金森病 MECP2 TH蛋白 SH-SY5Y细胞 6-OHDA 

分 类 号：R74[医药卫生—神经病学与精神病学] R392.1[医药卫生—临床医学]