基于Wnt/β-catenin通路探究槲皮素对腹膜透析大鼠腹膜纤维化的影响及机制OA

Objective To explore the effect and mechanism of quercetin(QCT)on peritoneal fibrosis(PF)in peritoneal dialysis(PD)rats based on the Wnt/β-catenin pathway.Methods PD rat model was constructed,and successfully modeled rats were stochastically classified into LiCl-N(LiCl,0 mg/kg),LiCl-L(LiCl,30 mg/kg)、LiCl-M(LiCl,60 mg/kg),LiCl-H(LiCl,90 mg/kg),PD,QCT-L(QCT,17.5 mg/kg),QCT-H(QCT,35 mg/kg),and QCT-H+LiCl-M(QCT,35 mg/kg+LiCl,60 mg/kg)groups.Another 12 rats served as the Ctrl group.Rats in the Ctrl and PD groups were given an equal amount of 0.9%sodium chloride solution at the same time and location by gavage once a day for 4 weeks.Peritoneal function was assessed using the peritoneal balance test.Changes in peritoneal tissue were observed by hematoxylin-eosin staining and peritoneal tissue fibrosis was detected by Masson staining.Levels of the inflammatory factors tumor necrosis factor(TNF)-α,interleukin(IL)-1β,and IL-6 were detected by enzyme-linked immunosorbent assay.α-smooth muscle actin(SMA),collagen type 1(CoL-1),and E-cadherin protein levels were detected by immunohistochemistry,and Wnt3a,low-density lipoprotein receptor-related protein 5(LRP5),lymphoid enhancer-binding factor 1(LEF-1),glycogen synthase kinase-(GSK)-3β,and β-catenin proteins were measured by Western blot.Results Compared with the LiCl-N group,rats in the LiCl-L groups showed no effects on serum creatinine(Scr)and blood urea nitrogen(BUN)levels,mass transfer of glucose(MTG),ultrafiltration volume(UF),Wnt3a,and β-catenin protein(P>0.05),Rats in the LiCl-M groups showed same effects,while Wnt3a,β-catenin prote were increased(P<0.05).Rats in the LiCl-H groups showed more Scr and BUN levels,mass transfer of glucose(MTG),and Wnt3a and β-catenin protein(P<0.05),while UF were decreased(P<0.05).Thus,60 mg/kg was selected as the LiCl concentration.Rats in the PD group shed peritoneal mesothelial cells compared with the Ctrl group's number of inflammatory cells,relative area of collagen fiber deposition,peritoneal thickness,Scr and BUN levels,MTG,peritoneal tissue TNF-α,IL-1β,IL-6,α-SMA,CoL-1,Wnt3a,LRP5,LEF-1,and β-catenin were all increased(P<0.05),while UF,E-cadherin,and GSK-3β proteins decreased(P<0.05).Compared with the PD group,rats in the QCT-L and QCT-H groups showed less mesothelial cell shedding,number of inflammatory cells,relative area of collagen fiber deposition,peritoneal thickness,SCr,BUN,MTG,peritoneal tissue TNF-α,IL-1β,and IL-6 levels,and α-SMA,CoL-1,Wnt3a,LRP5,LEF-1,and β-catenin proteins were decreased(P<0.05),while UF,E-cadherin,and GSK-3β were increased(P<0.05).Compared with the QCT-H group,rats in the QCT-H+LiCl-M group showed increased peritoneal tissue damage,increased number of inflammatory cells,relative area of collagen fiber deposition,peritoneal thickness,Scr and BUN levels,MTG,peritoneal tissue TNF-α,IL-1β,and IL-6 levels,and α-SMA,CoL-1,Wnt3a,LRP5,LEF-1,β-catenin proteins(P<0.05),and decreased UF,E-cadherin,and GSK-3β(P<0.05).Conclusions QCT alleviates PF in PD rats by inhibiting the inflammatory response and peritoneal mesothelial cell mesenchymal transition process,possibly acting via inhibition of the Wnt/β-catenin signaling pathway.