Research Paper Volume 15, Issue 22 pp 13384—13410

Figure 7. βOHB supplementation alleviates the progression of DKD. (A) Schematic diagram illustrating the animal experimental design. During the experiment, m/m mice were fed with CD, while db/db mice were fed with HFD. Eight-week-old male db/db mice were randomized to receive vehicle or βOHB, which was dissolved in drinking water at a final concentration of 100 mM. Mice were euthanized at six weeks after βOHB intervention. (B) Blood level of βOHB in mice from indicated groups (n = 5 per group). (C) Urinary ACR values of mice from indicated groups (n = 5 per group). (D) Negative correlation between serum βOHB level and ACR in db/db mice with vehicle and βOHB treatment. (E) Representative WB image showing the protein level of BDH1 in the kidneys of indicated groups. (F) Representative photomicrographs of H&E, Masson, IHC (IL-1β), and TUNEL staining showing the pathological changes in the kidneys of indicated groups. (G) Quantification of the fibrosis area in the kidneys of indicated groups (n = 4 per group). (H) Quantification of apoptosis-positive cells in the kidneys of indicated groups (n = 4 per group). All results are representative of three independent experiments. In B and C, values are presented as median (interquartile range). In E, G, and H, values are presented as mean ± standard deviation. Bar: 100 μm in F. Abbreviations: CD: control diet; HFD: high fat diet; ACR: albumin-to-creatinine ratio; BDH1: β-hydroxybutyrate dehydrogenase 1; βOHB: β-hydroxybutyrate; DKD: diabetic kidney disease; WB: western blot; H&E: hematoxylin and eosin; IHC: immunohistochemistry. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001.