SOCS3对IgA肾病患者IgA1刺激人肾小球系膜细胞增殖的影响及机制探讨.pdfVIP

Abstract ABSTRACT Objective： To investigate the effect of overpression SOCS3 on the proliferation of human mesangial cells stimulated by IgA1 from serum of patients with IgAN, and explore the possible mechanism. Method ： 1) Serum IgA1 from IgAN was isolated with jacalin affinity chromatography and Sephacryl S-200 HR gel filtration method then polymerized. 2) Human glomerular mesangial cell was transfected by overexpression SOCS3 adenovirus then stimulated by aIgA1. 3) Determine the time and concentration of aIgA1 and the transfection conditions, set up 4 experimental groups: a) Control group: HMC only with 10%FBS and 1% penicillin-streptomycin of 1640 culture medium for normal culture and timing change. b) IgA1 group: normal cultured HMC was stimulatedby aIgA1. c) IgA1+ Adv-EGFP group: normal cultured HMC was transfected with Adv-EGFP for 48 hours then stimulated by IgA1. d) IgA1+ Adv-SOCS3-EGFP group: normal cultured HMC was transfected with Adv-SOCS3-EGFP for 48 hours then stimulated by aIgA1. 4) MTT to detected cell proliferation, western blot and Real-time PCR to observe the expression of SOCS3, TLR4, TGF-β1protein and mRNA of each group. 5) The experimental data was expressed by x  s and T-test was performed by statistical software of IBM SPSS Statistics 20. The results in p0.05 showed that there was significant difference. Results ： 1) HMC was stimulated by aIgA1 for 12h, 24h and 48h, cell proliferation was promoted significantly after stimulated for 24h or 48h, and the difference between IV Abstract 0.25, 0.5, 1.0mg/ml group and normal group was statistically significant (p 0.05)