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. 2019 Mar 19;10:271. doi: 10.3389/fphys.2019.00271

FIGURE 6.

FIGURE 6

Empagliflozin increases V2R expression but decreases AQP2 expression in diabetic rat kidneys. (A) Representative immunoblot reacting with anti-V2R, anti-AQP2 and anti-p261-AQP2. (B) Densitometric analysis shows that renal expression of V2R protein in the empagliflozin-treated OLETF group was significantly increased compared with untreated OLETF or lixisenatide-treated OLETF rats. ∗P < 0.001 vs. other groups. (C) qRT-PCR shows that mRNA level of V2R was significantly increased in all OLETF rats than in LETO rats. Empagliflozin further increased V2R mRNA level compared with untreated OLETF control. ∗P = 0.001 vs. OLETF_C; p < 0.001 vs. OLETF_C, OLETF_L and OLETF_V. †P = 0.041 vs. OLETF_C. (D) Protein expression of whole renal AQP2 was more decreased in empagliflozin-treated diabetic rats than in untreated or lixisenatide-treated diabetic rats. ∗P = 0.026 vs. OLETF_C; P = 0.045 vs. OLETF_L. (E) Medullary AQP2 expression was significantly decreased in empagliflozin-treated OLETF rats compared with control or lixisenatide-treated OLETF rats. ∗P = 0.035 vs. OLETF_C; P = 0.014 vs. OLETF_L. (F) mRNA level of renal AQP2 was significantly higher in untreated OLETF control and lixisenatide-treated OLETF rats. ∗P < 0.001 vs. LETO; P = 0.003 vs. OLETF_E. †P < 0.001 vs. LETO and OLETF_E; P = 0.040 vs. OLETF_V. (G) The renal expression of pS261-AQP2 was significantly increased with empagliflozin treatment compared with no treatment or lixisenatide treatment in diabetic rats. ∗P < 0.001 vs. OLETF_C and OLETF_L. †P = 0.004 vs. OLETF_C; p = 0.007 vs. OLETF_L. (H) Representative renal sections immunostained with anti-AQP2. (I) In quantitative analysis of results for AQP2, empagliflozin-tread OLETF rats have lower distribution of AQP2 protein in apical membranes of kidneys. ∗P < 0.001 vs. other groups. †P < 0.001 vs. LETO and OLETF_E; P = 0.002 vs. OLETF_V. Magnification, ×200. (J) Representative immunofluorescence images show that the distribution of AQP2 in medullary area was relatively lower in empagliflozin- or voglibose-treated OLETF group than in other groups. Magnification, ×400. n = 8 per each group.