Figure 4.
Treatment with 2DG ameliorates cystic kidney disease in the long-term model without evident signs of toxicity. (A) Representative histologic pictures of treated cystic (Pkd1ΔC/floxTmCre) kidneys as defined in Figure 1A with 2DG or vehicle (NaCl) only reveal a marked reduction in cystic burden in the 2DG-treated kidneys. (B) Cystic index evaluated in kidneys from cystic (Pkd1ΔC/floxTmCre) animals treated with either 2DG or vehicle (NaCl) only as described in A. (C) Key inflammation genes were upregulated in untreated cystic (Pkd1ΔC/floxTmCre) compared with control kidneys. With 2DG, a trend of reduction (CD15 and CD68) or significant reduction (CD45) could be observed. Hprt expression was used as a normalizer for total cell number. (D) Immunohistochemistry confirmed a significant reduction in the number of CD45-positive cells per acquired field (350×435 μm) in 2DG–treated cystic kidneys compared with NaCl-treated tissues; ten fields were acquired in one renal medial section from three different animals. (E) 2DG treatment had no significant effect on the total body weight in either controls or mutant mice. (F) Alanine aminotransferase (ALT) and (G) aspartate aminotransferase (AST) values showed no liver toxicity induced by 2DG in any of the animals used. (H) During the standard hidden platform version of the water maze trials, 2DG-treated mice were faster to reach the hidden platform (escape latency) compared with the vehicle-treated littermates. (I) No difference was observed in the annulus crossing during the probe trial in the first day (first 30 seconds) of the reversal phase. Old goal (og) indicates the old platform position, old opposite (oo) specifies the new opposite position of the platform, and old left (ol) and old right (or) show the quadrant at the left and the right of the old goal, respectively. (J) Representative images of heart sections of NaCl- (n=6) and 2DG-treated (n=9) mice stained with Hematoxylin-Eosin showing absence of vacuolization caused by 2DG (upper panel). Representative brain coronal sections (5 µm) of NaCl (n=5) and 2DG-treated (n=6) mice stained with Hematoxylin-Eosin show absence of differences (lower panel). Cortical thickness (CC) and the hippocampal region (Hip) are indicated. NS indicates P≥0.05. Scale bars, 100 µm in J, upper panel; 1 mm in J, lower panel. *P<0.05 determined using the Mann–Whitney test for B and D, t test for C, and ANOVA for H and I; **P<0.01 determined using the Mann–Whitney test for B and D, t test for C, and ANOVA for H and I; ***P<0.001 determined using the Mann–Whitney test for B and D, t test for C, and ANOVA for H and I.