CMHX008,A Novel PPAR? Agonist,Improves Renal Function In HFD-induced Mice With Diabetic Renal Injury

Aim Classical thiazolidinediones,such as rosiglitazone,are commonly used in the treatment of type 2 diabetes,but accompanied by serious adverse reactions that limit their clinical use.Therefore,our previous study synthesized CMHX008 with partial PPAR? agonist ability.It was found to have insulin sensitization similar to rosiglitazone,showed better safety to heart,bone and body weight than rosiglitazone.Diabetic nephropathy is one of the main complications of diabetes.According to reports in the literature,rosiglitazone showed an anti-fibrotic effect on diabetic nephropathy,but the effect of CMHX008 on diabetic nephropathy is not clear.This study aimed to investigate the effect and mechanism of the novel PPAR? agonist CMHX008 on renal function of diabetic renal injury in high fat diet-fed mice and high glucose cultured-renal tubular epithelial HK-2 cells.Methods The urine and serum samples were collected from male C57BL/6 mice fed with HFD to detect related indexes by automatic biochemical analyzer,H&E staining was used to reveal the histomorphology of kidney,the renal collagen accumulation was assayed by Sirius red and Masson's trichome staining,the mRNA levels of ?-SMA,E-cad,collagen I and fibronectin in kidney were assessed by quantitative real time RT-PCR;HK-2 cells cultured with 30 mM/L D-(+)-Glucose,then treated with Rosi and C.Cell Counting Kit-8(CCK-8)was used todetectcells viability,immunoblotting was used to detect the expression of fibrogenic markers in HK-2 cells,quantitative real time PCR(qRT-PCR)was used to detect fibrosis-related mRNA levels,wound healing assay and migration assay were used to detect the migration and invasion ability of HK-2 cells.Results HR group and HC group showed a decrease of Upro/Cr,and HC group showed significantly decreases of Scr and serum BUN;H&E staining revealed rosiglitazone and CMHX008 could improve HFD induced obvious tubular injury;Sirius red and Masson's trichrome staining revealed less accumulation of collagen in HR and HC group than in HV group;Compared with HV group,kidney mRNA levels of HR group and HC group showed down-regulated expression of ?-SMA,collagen I,and fibronection,and up-regulated expression of E-cadherin.CCK-8 analysis showed that 3?mol/L Rosi and 3?mol/L C had no obvious cytotoxicity to HK-2 cells;immunoblotting showed that Rosi and C could reverse the up-regulation of p-PPAR?(ser273)and reduce TGF-?1,?-SMA protein levels in HK-2 cells under high glucose conditions,and E-cad protein expression levels were increased;wound healing assay and migration assay showed that Rosi and C could decrease the invaded distance and migrated cells of HG-induced HK-2 cells.Conclusion Treatment of rosiglitazone and CMHX008 can effectively improve renal function and attenuate renal fibrosis in high fat diet-induced diabetic mice,and inhibit hyperglycemia induced fibrogenesis in human renal tubular epithelial HK-2 cells,which may be possibly related to inhibition of p-PPAR?(Ser273)phosphorylation.