| Objective: By establishing acute renal ischemia-reperfusion injury rat model,setting total panax notoginseng saponins(PNS)as intervention drug,and comparing with wnt pathway agonist or inhibitor,serum creatinine,urea nitrogen,SOD and MDA in renal tissue homogenates,the expression of wnt/?-catenin pathway related proteins in renal pathology and tissues are observed at different time points,in order to investigate the possible mechanism of PNS in renal protection.Method: 90 male Sprague-Dawley(SD)rats were randomly and equally divided into five groups,namely control group,ischemia-reperfusion(IR)group,panax notoginseng saponins(PNS)group,wnt agonist(agonist)group and wnt inhibitor(inhibitor)group.Experimental intervention: 1.Control group: daily tail intravenous injection of normal saline in the same amount to PNS group 3d before experiment,right kidney excision after anesthesia,abdomen-closing after 30 min exposure of left kidney;2.IR group: the same preparation with control group before anesthesia,right kidney excision,model established by restoring blood flow 30 min after left renal artery clipping;3.PNS group: daily tail intravenous injection with PNS(150mg·kg)3d before experiment,other procedures the same with IR group;4.Agonist group:daily q12 h tail intravenous injection of wnt agonist recombinant DKK2 protein(DKK2-C2,20ug·kg-1)3d before experiment,other procedures the same with IR group;5.Inhibitor group: daily q12 h tail intravenous injection of wnt inhibitor recombinant endostatin(Endostar,1.5mg·kg-1)3d before experiment,other procedures the same with IR group.The Scr,Bun,SOD and MDA in in renal tissue homogenates were extracted from 6 rats in respective group at 24 h,48h and 72 h after restoring blood perfusion.Tubular injury scoring was calculated by observing renal pathological changes under microscope in HE staining,and the expressions of wnt4 and ?-catenin in renal tissues were detected via immunohistochemical method andWestern blot.Results:1.Scr and BUN: except control group,IR group,PNS group peaked at 24 h,and inhibitor group at 48h;PNS group and agonist group were lower than IR group at different time points(P<0.05);inhibitor group was higher than IR group at different time points(P<0.05).2.Renal HE staining and tubular injury scoring: except control group,renal pathological damages in various degrees were observed in these groups.The tubular damage scores in IR group,PNS group and agonist group peaked at 24 h,and PNS group was lower than IR group in all different time points(P<0.05);The tubular damage scores in inhibitor group peaked at 48 h,which was significantly higher than that of IR group in all time points(P<0.05).3.Oxidative stress indicators of MDA and SOD: except the control group,MDA peaked in all groups at 24 h,but SOD declined the lowest value.Comparing with IR group,the MDA of PNS group and agonist group was significantly reduced at 24 h and 48h(P<0.05),while SOD was significantly increased(P<0.05);the MDA in inhibitor group was significantly higher than that of IR group at all time points(P<0.05),while SOD was noticeably lower than that of IR group(P<0.05).4.Wnt4 detection result: Immunohistochemistry showed that Wnt4 was no significant expression in the control group,the sites with expression were the same in other groups,that is,wnt4 was positive in renal tubular epithelial cell cytoplasm,glomerulus no significant expression,and the tendency was consistent with Western blot results.Except for control group,wnt4 expressions in all groups peaked at 24h;the expression in PNS group and agonist group was higher than IR group at all time points(P<0.05);the expression in inhibitor group was lower than IR group at all time points(P<0.05).5.?-catenin detection result: Immunohistochemistry showed that ?-catenin was positive in renal tubular epithelial cell membrane in the control group and ?-catenin was positive in the cytoplasm and membrane of renal tubular epithelial cells in othergroups,glomerulus no significant expression,and the tendency was consistent with Western blot results.Except for control group,?-catenin expressions in all groups peaked at 24h;the expression in IR group,PNS group and inhibitor group peaked at72h;comparing to IR group,the expression in PNS group and agonist group was higher at all time points(P<0.05);the expression in inhibitor group peaked at 48 h,which was significantly lower than in IR group at all time points(P<0.05).Conclusion:1.The renal protective mechanism of panax notoginseng total saponins may be related to up regulation of wnt/?-catenin pathway proteins.As for wnt agonist,it also promote the recovery in ischemia-reperfusion injuried kidneys.2.Wnt/?-catenin signaling pathway is involved in repair process after renal ischemia-reperfusion.Renal injury is aggravated via wnt inhibitor. |
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