To Explore The Effect Of SOCS3 On The Renal Injury Of IgA Nephropathy Mouse And Relative Mechanism

Objective:To investigate the expression of cytokine signal transduction inhibitor 3(SOCS3) and Toll like receptor 4(TLR4) in IgA nephropathy(IgAN) mice,and explore the possible roles and mechanisms of SOCS3 in renal injury of Ig AN mice through SOCS3-lentivirus in vivo transfections.1. BALB / c mice as the research object, adopting of the immunity-inducing method: bovine serum albumin(BSA) + lipopolysaccharide(LPS) + carbon tetrachloride(CCL4) to establish IgA nephropathy mouse model.2. Establish SOCS3-overexpression mouse model through the lentivirus transfection technique in vivo, and set up four experimental groups: normal control group, IgAN group, IgAN + negative virus solution, IgAN + SOCS3 lentivirus group.3. Two weeks after in vivo transfection, we tested the blood serum creatinine, serum albumin, alanine aminotransferase, and kept the mice kidney for HE and IgA immunofluorescence.we assessed the damage of renal pathology by Katafuch scoring standard.4. Immunohistochemical detection of mice kidney SOCS3, TLR4, MCP-1, TGF-?1 expression and localization, Western blot detection of renal cortex SOCS3, TLR4, MCP-1, TGF-?1 expression, real-time fluorescence quantitative PCR detection the mRNA expression of renal cortex SOCS3, TLR4, MCP-1, TGF-?1.5. Detecting the level of blood serum IgA though ELISA.Result:1. In the modeling process of normal mice in good spirits, while the mice in model group appeared coarse hair, loss of appetite, listlessness and other symptoms. In blood biochemical tests,we find IgAN plasma albumin was significantly lower than the normal control group(P <0.05), IgAN + SOCS3 group compared to IgAN group, IgAN + negative viruses were significantly higher(P <0.05). IgAN group blood alanine aminotransferase and creatinine was significantly higher than the normal control group(P <0.05), IgAN + SOCS3 group compared to IgAN group, IgAN + virus negative group was significantly decreased(P <0.05).2. The normal control group showed normal glomerular structure in HE,while the IgAN group and IgAN + virus negative group showed obvious mesangial cell proliferation, inflammatory cell infiltration, tubular atrophy, interstitial fibrosis. Compared with normal control group, IgAN group pathology score was significantly increased, the difference was statistically significant(P <0.05). IgAN + SOCS3 group compared with IgAN group, the pathological score was significantly decreased, the difference was statistically significant(P <0.05).3.IgA immunofluorescence of normal control group was negetive, while in the IgAN group, IgAN + virus negative group, IgAN + SOCS3 group were seen grainy and lumpy IgA deposits in glomerular mesangial area. Immunofluorescence brightness was no significant difference among the three groups.4. The results of immunohistochemistry: SOCS3, TLR4 is mainly expressed in the renal tubules, little expression in glomeruli, MCP-1, TGF-?1 mainly expressed in glomeruli, little expression in renal tubules. The expression of SOCS3, TLR4, MCP-1, TGF-?1 in IgAN group was significantly higher than the control group, The expression of SOCS3 in IgAN + SOCS3 group was significantly increased when compared with IgAN group, while the expression of TLR4 was decreased,and the expression of MCP-1, TGF-?1 was no significant difference(P> 0.05).5. The mRNA and protein expression of SOCS3,TLR4, MCP-1, TGF-?1 in normal control group was significantly lower than IgAN group. The mRNA and protein expression of SOCS3 in IgAN + SOCS3 group was significantly increased compared with IgAN group, while the amount of TLR4 protein and mRNA expression was significantly decreased, and the protein and mRNA expression of MCP-1 was no significant difference.the protein expression of TGF-?1 decreased significantly(P <0.05), but the mRNA expression levels had no significant difference.6. ELISA:We find serum IgA concentration in IgAN group was significantly higher than the control group(P <0.05), IgAN + SOCS3 group compared to IgAN group, IgAN + negative virus was significantly decreased(P <0.05).Conclusion:1. Adoption of immune induction method successfully constructed a mouse model of IgA nephropathy.2. There are low expression of SOCS3, TLR4, MCP-1 and TGF-?1 in kidney tissue of the control group, while SOCS3, TLR4, MCP-1 and TGF-?1 expression are increased in the IgAN model group.3. Though SOCS3-lentivirus transfection in vivo, the expression of SOCS3 is increased in IgAN mice renal tissue, and may reduce renal pathological damage by down-regulating the expression of TLR4, thus play a protective role in Ig AN renal damage.