Research Paper Volume 15, Issue 22 pp 13384—13410

Figure 2. AAV9-mediated BDH1 ectopic expression in kidneys alleviates the progression of DKD. (A) Schematic diagram illustrating the animal experimental design. During the experiment, m/m mice were fed a control diet (CD), while db/db mice were fed an HFD. Six-week-old male db/db mice were randomized to receive tail vein injection of AAV9-Bdh1-GFP (3.40E+12 vg/mL) or vector (1.90E+13 vg/mL). Mice were euthanized at 11 weeks after AAV9 injection. (B) Urinary ACR values of mice in the indicated groups (n = 5 in m/m group, n = 4 in vector, and AAV-Bdh1-GFP injected db/db group). (C) Representative renal fluorescent images of the mice 11 weeks after caudal vein delivery of AAV-Bdh1-GFP revealing GFP expression in the kidneys. (D) Representative WB image showing the protein level of BDH1 in the kidneys of indicated groups. (E) Representative photomicrographs of H&E, Masson, IHC (IL-1β), and TUNEL staining showing the pathological changes in the kidneys of indicated groups. (F) Quantification of the fibrosis area in the kidneys of indicated groups (n = 4 per group). (G) Quantification of apoptosis-positive cells in the kidneys of indicated groups (n = 4 per group). All results are representative of three independent experiments. Values are presented as mean ± standard deviation. Bar: 100 μm in C and E. Abbreviations: CD: control diet; HFD: high fat diet; ACR: albumin-to-creatinine ratio; AAV9: adeno-associated virus 9; BDH1: β-hydroxybutyrate dehydrogenase 1; DKD: diabetic kidney disease; WB: western blot; H&E: hematoxylin and eosin; IHC: immunohistochemistry. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001.