Protective Effect And Mechanical Study Of Tacrolimus On Adriamycin-induced Nephropathy In Rat

Objective: This study investigated the protective effect of tacrolimus(FK506)on the kidneys of adriamycin-induced nephropathic rats and the effects on the Notch signalling pathway,also the protective effects of tacrolimus and triptolide(TP)on kidneys were compared.Methods : Adriamycin-induced nephropathy model was established in rats,which were randomly divided into an adriamycin nephropathy group(ADR group,ADR 6.5 mg/kg),triptolide-treated ADR group(TP group,ADR 6.5 mg/kg+TP 200 mg/kg/d),tacrolimus-treated ADR group(FK506 group,ADR 6.5 mg/kg+FK506 0.5 mg/kg/d)and Control group(N group).Rats in the ADR group,TP group and FK506 group were fixed respectively.The nephropathy model was induced via two tail vein injections of Adriamycin.After one week,the rats were collected 24-hour urine with metabolic cages respectively to test twenty-four-hour urinary protein.Compared with the N group,twenty-four-hour urinary protein in the ADR group,the TP group and the FK506 group was statistically significant as a successful model.The TP and FK506 groups received drug interventions for 8 weeks.Twenty-four-hour urinary protein was measured on weeks 4,6 and 8.Rats were sacrificed on the 8th week.Blood biochemistry was measured using an automatic biochemical analyser,and renal histology was observed under light and electron microscopes.Expression of Fkbp12,Notch1 and Hes1 in renal tissue was detected using real-time PCR and Western blot.Results :(1)Twenty-four-hour urinary protein increased significantly in ADR rats compared to control rats.The 24-hour urinary protein and blood biochemistries of TP rats and FK506 rats were lower than those of ADR rats.The 24-hour urinary protein and total cholesterol(TC)of FK506 rats were lower than those of TP rats.(2)The histological changes in ADR rats included hyperplasia of the glomerular mesangial and endothelial cells,increased matrix deposition,and an expanded mesangial area.The histological changes in TP and FK506 rats were more subtle than those in ADR rats.(3)Notch1 and hes1 mRNA and protein expression in ADR rats were higher than those in control rats(P <0.05).Notch 1 and hes1 mRNA and protein expression in TP and FK506 rats were lower than those in ADR rats(P <0.05).Notch 1 and hes1 mRNA and protein expression in FK506 rats were lower than those in TP rats(P<0.05).There was no statistical difference in the expression of Fkbp12 in each group.Conclusion:(1)This study demonstrated that adriamycin-induced nephropathy was associated with activation of the Notch signalling pathway.(2)After the intervention of triptolide and tacrolimus,the expression of Notch1 and Hes1 was relatively decreased,suggesting that Triptolide and tacrolimus may play the role of renal protection by inhibiting abnormal expression of Notch signals.(3)Compared with the TP group,the inhibitory effect of tacrolimus on the Notch pathway was more obvious,and the 24 h urine protein and cholesterol(TC)were lower than those in the TP group,which showed that tacrolimus exhibited superior efficacy compared to triptolide in the treatment of adriamycin-induced nephropathy.