Effect Of Jieduquyuziyin-prescription On MeCP2 Gene Expression In Jurkat Cells

Objective Methyl CpG binding protein 2(MeCP2)protein was used as a molecular target.DNA methylation levels were tested by genomic DNA methylation kits.Real-time PCR was employed to test the expression of MeCP2 mRNA.Western blot and flow cytometry were performed to detect the MeCP2 protein expression in the Jurkat cell nucleus.To investigate the effect of Jieduquyuziyin prescription treated Rat Serum on the expression of MeCP2 protein and mRNA in Jurkat cells,and to explore the mechanism of Jieduquyuziyin prescription on the mechanism of SLE.Methods1.Jurkat cells(human peripheral blood leukemia T cells)are resuscitated and cultured in the logarithmic growth phase.Then drug-containing serum are prepared.MTS assay was used to screen the optimal concentration of serum-containing drugs.They are prepared for follow-up experiments.2.Jurkat cells are placed in 6-well plate,and the cells are randomly divided into five groups:①Traditional Chinese medicine(Jieduquyuziyin-prescription)serum group;②Western medicine(prednisone acetate)group;③Combination group;④Blank group(distilled water);⑤ Fetal bovine serum group.Jurkat cells are harvested after 48 hours of incubation.3.DNA methylation levels were tested by genomic DNA methylation kits in Jurkat cells.Real-time PCR is employed to test the expression of MeCP2 mRNA.Western blot and flow cytometry are performed to detect the MeCP2 protein expression in the Jurkat cell nucleus.Results1.The DNA methylation level,MeCP2 mRNA and protein expression level increased significantly in the Jieduquyuziyin prescription groups and prednisone acetate groups compared with that of the control group as detected by Real-timePCR,Western blot and FCM(P<0.05).2.The combined group’s protein expression level was statistically significant(P<0.05),while the expression level of DNA methylation and MeCP2 mRNA did not change significantly(p>0.05).Conclusion1.The level of DNA methylation,MeCP2 mRNA and protein in Jurkat cells were significantly increased by the Jieduquyuziyin prescription,and the expression of MeCP2 gene and protein were up-regulated.The effect was consistent with prednisone acetate group.2.The role of Jieduquyuziyin prescription may be as same as prednisone acetate,the regulatory mechanism mainly by increasing the expression of MeCP2 mRNA levels,activation and upregulation of MeCP2 protein,so that DNA methylation level was in a normal dormant state,effectively inhibiting the immune the occurrence of SLE.Patients with immune disease have a potential therapeutic effect.