Study On The Effect Mechanism Of Qidi Gushen Tablets On Idiopathic Membranous Nephropathy Based On NF-κB/NLRP3 Signal Transduction Pathway

Objective:1.Observe the 24 h urine protein quantification,serum biochemistry and kidney pathology in idiopathic membranous nephropathy(IMN)model rats to clarify the efficacy of Qidi Gushen tablets on IMN model rats.2.Observe the effect of Qidi Gushen tablets on the activation of NF-κB/NLRP3 signal transduction pathway in renal tissue of IMN rats,to further explore the mechanism of Qidi Gushen tablets in the treatment of IMN.Methods:This experiment uses cationized bovine serum albumin(C-BSA)to replicate a rat model of idiopathic membranous nephropathy.Take 60 healthy male SD rats,and reserve9 in blank groups.After successful modeling,the rest rats were randomly divided into model group,tripterygium wilfordii polyglycoside tablets group,low-dose group of Qidi Gushen tablets(hereinafter referred to as low-dose group of traditional Chinese Medicine),medium dose group of Qidi Gushen tablets(hereinafter referred to as medium dose group of traditional Chinese Medicine)and high-dose group of Qidi Gushen tablets(hereinafter referred to as high-dose group of traditional Chinese Medicine),a total of 6 groups.Each administration group was given the corresponding drug suspension by gavage,while the blank group and model group were gavage with the same amount of normal saline for 4weeks.Observe the 24 h urine protein quantification dynamically and the levels of serum albumin,total protein,alanine aminotransferase,aspartate aminotransferase,triglycerides,total cholesterol,blood creatinine,and urea nitrogen after administration.HE,MASSON staining and electron microscope were used to observe the pathological changes of rat kidney.WB and RT-PCR were used to detect the expression of NF-κB/NLRP3 signal transduction pathway related factors NF-κB-p65,p-NF-κB-p65,IκBα,p-IκBα,NLRP3,ASC and Caspase-1 in renal tissue.Result:1.24 h urine protein quantification.There was no difference between the groups before modeling,and they were comparable;after modeling,before the administration,proteinuria of the model group and each administration group was significantly higher than that of the blank group(P<0.01);after 2 weeks of administration,each administration group proteinuria decreased but there was no statistical difference compared with the model group(P>0.05);after 4 weeks of administration,compared with the model group,the proteinuria of the tripterygium wilfordii polyglycoside tablets group and the high,middle and low dose groups of Chinese medicine were significantly reduced(P<0.01),there was a decrease in the low dose group of traditional Chinese medicine(P<0.05).2.Serum albumin and total protein.Compared with the blank group,the serum ALB and TP of the model group were significantly reduced(P<0.01);compared with the model group,the serum ALB and TP of each administration group increased with significant difference(P<0.01).3.Blood lipids in IMN rats.Compared with the blank group,the serum TC and TG of the model group were increased with significant difference(P<0.01);compared with the model group,the TC and TG of the tripterygium wilfordii polyglycosides tablets group,the Chinese medicine high and medium dose groups were decreased,there was a difference.4.Pathology of kidney tissue.Observation under light microscope: in the model group,the basement membrane of glomerulus thickened diffusely,and the deposition of subepithelial furophilic protein could be seen by MASSON staining.There were pathological changes in the glomerular basement membrane in the other four groups: after administration,compared with the model group,the pathological changes in the high and middle dose groups and tripterygium wilfordii polyglycosides tablets group were relatively mild,the glomerular basement membrane was locally thickened,and a small amount of subepithelial furophilic protein deposition could be seen by MASSON staining.Electron microscopic observation: in the model group,the basement membrane thickened diffusely,the electronic dense deposited under the epithelium,and the foot process of epithelial cells fused diffusely.There were pathological changes in the glomerular basement membrane in the other four groups: after administration,compared with the model group,the pathological changes in the high and middle dose groups and tripterygium wilfordii polyglycosides tablets group were relatively mild,the basement membrane was slightly inevenly thickened,and a small amount of electronic densification could be seen,and foot processes are fused.5.The expression of NF-κB-p65,IκBα,NLRP3 and Caspase-1 mRNA in rats kidney tissues: compared with the blank group,the expression of NF-κB-p65 mRNA in the model group was significantly increased(P<0.01),NLRP3 mRNA was increased(P<0.05),IκBαmRNA expression was decreased(P<0.05),but the expression of Caspase-1 mRNA had no significantly change(P>0.05);compared with the model group,the expression of NF-κB-p65 mRNA in the high and medium dose groups of traditional Chinese medicine decreased significantly(P<0.01),and the NF-κB-p65 mRNA in the tripterygium wilfordii polyglycosides tablets group decreased(P<0.05).At the same time,the NLRP3 mRNA in the Chinese medicine middle and high dose group and the tripterygium wilfordii polyglycosides tablets group was reduced(P<0.05),the IκBα mRNA in the Chinese medicine middle dose group was significantly increased(P<0.01).6.The expression of p-NF-κB-p65,p-IκBα,NLRP3 and ASC protein in renal tissues of rats: compared with the blank group,the expressions of p-NF-κB-p65,p-IκBα,NLRP3 and ASC protein in the model group were significantly increased(P<0.01);compared with the model group,the expression of p-NF-κB-p65,NLRP3 and ASC protein in the the Chinese medicine high and medium dose groups and tripterygium polyglycosides tablets group was significantly reduced,and the difference was significant(P<0.01),the expression of p-IκBα protein in the high and medium dose groups of Chinese medicine was significantly reduced(P<0.01),the expression of p-IκBα protein in the tripterygium wilfordii polyglycosides tablets group decreased(P<0.05).Conclusion:1.Qidi Gushen tablets can reduce 24 hU-TP,increase ALB,TP,and reduce TC and TG levels in IMN model rats.2.Qidi Gushen tablets can reduce the pathological damage of the kidney in IMN model rats,and inhibit the deposition of glomerular basement membrane immune complexes and the thickening of basement membrane in IMN model rats.3.Qidi Gushen tablets may inhibit the renal immune inflammatory response and alleviate the renal injury of IMN by inhibiting the activation of NF-κB/NLRP3 signal transduction pathway in IMN model rats.