Ruboxistaurin attenuates diabetic nephropathy via modulation of TGF-[beta]1/Smad and GRAP pathways

Objective To investigate whether ruboxistaurin (a selective PKC-[beta] inhibitor) mediates renoprotective effect via interference with TGF-[beta]1/Smad-GRAP cross-signalling. Method Diabetes was induced in rats by a single intraperitoneal injection of streptozotocin (55 mg/kg). Then, the diabetic rats were treated with ruboxistaurin (10 mg/kg, p.o) for 6 weeks. Valsartan (15 mg/kg, p.o) was used as a positive control. After 6 weeks of treatment, diabetic nephropathy biomarkers were assessed. TGF-[beta]1, Smad2, and Smad3 mRNA and protein levels were detected using qPCR and western blot analysis. Key findings Data showed that serum creatinine, kidney/body weight ratio and urinary albumin excretion significantly increased in diabetic rats. These changes were significantly attenuated by treatment with ruboxistaurin. A significant up-regulation of TGF-[beta]1, Smad2 and Smad3 mRNA expression was observed in diabetic rats, which was alleviated by administration of ruboxistaurin. Furthermore, immunoblotting showed a significant improvement in protein levels of TGF-[beta]1 (P < 0.01), Smad2/3 (P < 0.01) and p-Smad3 (P < 0.001) in diabetic rats treated with ruboxistaurin compared to untreated. Importantly, the reduction in GRAP protein expression in diabetic kidney was prevented by treatment with ruboxistaurin. Conclusion These data suggest that the renoprotective effect of ruboxistaurin is possibly due to down-regulation of TGF-[beta]1/Smad pathway and normalization of GRAP protein expression.