Genistein alleviates renal injury in rats with membranous nephropathy by inhibiting TLR4/NF-κB signaling pathway
Objective It is to investigate the effect of genistein on renal injury in membranous nephropathy(MN)rats and toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway.Methods Eight of forty-five SD rats were randomly selected as normal group,and the other 37 rats were injected with cationized bovine serum albumin(C-BSA)by tail vein to establish MN models.32 successfully modeled rats were randomly divided into model group,genistein group,Resatorvid group and genistein +Resatorvid group,with8 rats in each group.The genistein group was given genistein30 mg/kg intraperitoneally,the Resatorvid group was given Resatorvid 10 mg/kg intraperitoneally,and genistein + Resatorvid group was given genistein 30 mg/kg and Resatorvid 10 mg/kg intraperitoneally,all once daily,and intervened for 4 weeks.The rats in each group was given intraperitoneal injection once a day for 4 weeks.The renal function indexes[24 h urinary protein(24 h UTP),blood urea nitrogen(BUN),serum creatinine(SCr),albumin(ALB)]were detected;the pathologic mor-phology,collagen deposition and cellular ultrastructure of renal tissues were observed by HE staining,Masson staining and transmission electron microscopy;immunoglobulin G(IgG)deposition in renal tissues was observed by immunofluorescence staining;the levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in renal tissues were detected by ELISA;the expressions of TLR4,NF-κB p65,and transforming growth factor-β1(TGF-β1)mRNA were detected by RT-PCR,and the expressions of proteins related to TLR4/NF-κB signaling pathway in renal tissues were detec-ted by Western blot.Results Compared with the normal group,the levels of 24 h UTP,BUN,SCr of rats were significantly higher,while the levels of ALB was significantly lower in the model group(all P＜0.05);there were pathological changes such as glomerular volume increase,tubular cell vacuolation,interstitial inflammatory cell infiltration,mass collagen depo-sition in interstitial region and glomerular,and the collagen volume fraction(CVF)was significantly increased(P＜0.05);there were ultrastructural changes such as glomerular cells swollen,foot process fused or disappeared,basement membrane irregularly thickened,mass electron dense deposition under the epithelial cells,mass IgG deposition;the levels of IL-1β,IL-6,and TNF-α,and the relative expressions of TLR4,NF-κB p65,and TGF-β1 mRNA and the relative ex-pressions of TLR4,NF-κB p65,TGF-β1,collagen type Ⅰ(Col-Ⅰ),type Ⅲ collagen(Col-Ⅲ),and p-Smad2/3 pro-teins in renal tissues were significantly increased(all P＜0.05),and the relative expression of Smad7 protein in renal tis-sue was significantly decreased(P＜0.05).Compared with the model group,the level of 24 h UTP,BUN,SCr were sig-nificantly decreased,while the levels of ALB were significantly increased in genistein group,Resatorvid group and genistein+ Resatorvid group(all P＜0.05);the pathological changes in renal tissue and ultrastructural changes in cells were signifi-cantly improved,CVF and IgG deposition were significantly decreased(all P＜0.05);the levels of IL-1β,IL-6,TNF-α and the relative expressions of TLR4,NF-κB p65,TGF-β1 mRNA and the relative expressions of TLR4,NF-κB p65,TGF-β1,Col-Ⅰ,Col-Ⅲ,and p-Smad2/3 proteins in renal tissues were all significantly reduced(all P＜0.05),and the relative expressions of Smad7 protein were all significantly increased(all P＜0.05).Compared with the genistein group and Resatorvid group,the renal injury was milder and the improvements of every indexes were more significant in the genistein +Resatorvid group(all P＜0.05).Conclusion Genistein can obviously improve improve renal function and alle-viate kidney injury in membranous nephropathy rats,its mechanism may be related to inhibiting TLR4/NF-κB signaling pathway,alleviating inflammatory response and renal fibrosis.
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