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. 2020 Feb 18;11(2):211. doi: 10.3390/genes11020211

Figure 1.

Figure 1

Inhibition of ACE suppresses DPP-4 and associated TGFβ signaling in diabetic kidneys (A) Quantitative analysis of DPP-4 mRNA expression by real time PCR using specific primers in the kidney of control, DM, DM+ACEi, DM+combination and DM+ARB treated mice. N = 6 were analyzed in each group. 18S was used as internal control to normalize the expression data. (B) Western blot analysis of DPP-4, TGFβR1, p-smad3, smad3, FSP-1, αSMA, Colla1a and fibronectin (FN) in the kidney of control, DM, DM + ACEi, DM + combination treatment and DM + ARB treated diabetic mice. Representative blots are shown. Quantification of DPP-4, TGFβR1, smad3 phosphorylation, FSP-1, αSMA, Colla1a and FN by densitometry. The data were normalized by β-actin. N = 5 were analyzed in each group. (C) Co-immunofloroscence analysis of DPP-4/CD31 and DPP-4/ αSMA in the kidney of control, DM, DM + ACEi, DM + ACEi + AcSDKP and DM + ARB, the representative pictures are shown. Scale bar 50 µm. DPP-4 FITC (green) labeled whereas, CD31 and αSMA are rhodamine labeled and DAPI blue. N = 5 were analyzed in each group. (D) DPP activity analysis by fluorimeter in kidney homogenate of control, DM, DM + ACEi, DM + combination and DM + ARB treated mice. N = 6 were analyzed in each group. (E) DPP activity analysis in the plasma of control, DM, DM + ACEi, DM + combination and DM + ARB treated mice. N = 6 were analyzed in each group. Data in the graph are presented as mean ± SEM. One-way Anova Tukey test was performed for calculation of statistical significance. C = control (non-diabetic), DM = diabetic group, combination = (ACEi + AcSDKP), Colla1 = collagen I, FN = fibronectin.