Effect Of Shenyuan Granules On Calcium-phosphorus Metabolism By MiR-199b-5p/klotho/FGF23 Signaling Pathway In Diabetic Nephropathy

Objective:Research the effect of Shenyuan granules on calcium-phosphorus metabolism by mi R-199b-5p/Klotho/FGF23 signaling pathway in diabetic nephropathy.Method:In vivo study,STZ-induced mice models were randomly divided into Shenyuan granules group [6.17/(kg·d)],Irbesartan group [0.03 g/(kg· d] and model control group,with the blank control group(n=10)established independently.Corresponding drugs were given by gavage for 8 weeks.And fasting blood glucose(FBG),24-hour microalbuminuria(24h m ALB),blood urea nitrogen(BUN),serum creatinine(SCr),calcium(Ca),phosphor(P)were detected.Moreover,the RT-q PCR was used to detect mi R-199b-5p and the m RNAs of Klotho,FGFR-1 and EGR-1.The Western-blot and Immunohisto--chemistry was used to detect the protein of Klotho,FGFR-1,EGR-1 and Na Pi2 c.Result:In vivo study,microarray-based analyze screened mi R-199b-5p as the potential research object.In the biochemical examinations,levels of the FBG,serum P,SCr and 24 h m ALB in model control group,Shenyuan granules group and irbesartan group were all higher than that in blank control group(P<0.05).And the serum P and SCr in Shenyuan granules group and irbesartan group were lower than that in model control group(P<0.05).P and SCr in Shenyuan granules group were lower than that in irbesartan group(P < 0.05).There were no significant differences in serum urea nitrogen in each group(P>0.05).In the result of RT-q PCR,compared with the blank control group,the expressions of mi R-199b-5p and EGR-1 were up-regulated in model control group,Shenyuan granules group and irbesartan group(P<0.05).The expressions of mi R-199b-5p and EGR-1 in Shenyuan group and irbesartan group were lower than that in model control group(P<0.05).The expressions of Klotho and FGFR-1 in model control group,Shenyuan granules group and model control group were lower than that in blank control group(P<0.05).And the levels of Klotho and FGFR-1 in Shenyuan granules group and irbesartan group were higher than that in the control group(P<0.05).The expression levels of Klotho and FGFR-1 in Shenyuan granules group were higher than that in irbesartan group(P<0.05).In the result of western blot and immunohistochemistry,compared with the blank control group,the expressions of Na Pi2 c and EGR-1 were up-regulated in model control group,Shenyuan granules group and Irbesartan group(P<0.05).The expressions of Na Pi2 c and EGR-1 in Shenyuan group and irbesartan group were lower than that in model control group(P<0.05).The expression level of Na Pi2 c in Shenyuan granules group was lower than that in irbesartan group(P<0.05).The expressions of Klotho and FGFR-1 in model control group,Shenyuan granules group and model control group were lower than that in blank control group(P < 0.05).And the expressions of Klotho and FGFR-1 in Shenyuan granules group and irbesartan group were higher than that of the model control group(P<0.05).The expression levels of Klotho and FGFR-1 in Shenyuan granules group were higher than that in irbesartan group(P<0.05).In the result of ELISA,compared with the blank control group,the expressions of FGF23 were up-regulated in model control group,irbesartan group and Shenyuan granules group(P<0.05).And the expressions of FGF23 in model control group was higher than that in Shenyuan granules group and irbesartan group(P<0.05).There were no significant differences in FGF23 between Shenyuan granules group and irbesartan group(P>0.05).Conclusion:1.Shenyuan granules can effect renal function and calcium--phosphorus metabolism abnormality in DN model.And the effect of Shenyuan granule on serum phosphorus and serum creatinine in model group is superior to that of irbesartan.2.Shenyuan granules can affect the RNA expression of mir-199b-5p /Klotho/FGF23 pathway in the DN model.And the effect of Shenyuan granule on Klotho and FGFR-1 in DN model is superior to that of irbesartan.3.Shenyuan granules can affect the protein expressions of mir-199b-5p /Klotho/FGF23 pathway in the DN model.And the effect of Shenyuan granules on the expression of Klotho,FGFR-1 and Na Pi2 c is superior to that of irbesartan.