Renoprotective effects of brown adipose tissue activation in diabetic mice

Figures

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  Figure 1. Effects of CL316,243 on A, blood glucose and B, serum lipids in diabetic mice. Data are the mean ± SEM (n = 7‐9 per group). *P < 0.05, **P < 0.01, ***P < 0.001 compared with the control group; †† P < 0.01, ††† P < 0.001 compared with the diabetic control (DM‐Con) group. DM + CL, diabetic mice treated with CL316,243; HDL‐C, high‐density lipoprotein cholesterol; LDL‐C, low‐density lipoprotein cholesterol; TG, triglycerides

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  Figure 2. Effects of CL316,243 on 24‐h urinary A, albumin excretion and B, 8‐hydroxydeoxyguanosine (8‐OHdG) concentrations in diabetic mice. Data are the mean ± SEM (n = 7‐9 per group). *P < 0.05, **P < 0.01, ***P < 0.001 compared with the control group; † P < 0.05, †† P < 0.01, ††† P < 0.001 compared with the diabetic control (DM‐Con) group. DM + CL, diabetic mice treated with CL316,243

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  Figure 3. Effects of CL316,243 on relative mRNA expression of A, mitochondrial and thermogenic genes in epididymal adipose tissues (EP) and B, Ucp1 in interscapular brown adipose tissue (BAT) in diabetic mice. Control, non‐diabetic mice; DC‐Con, diabetic control mice; DM + CL, diabetic mice treated with CL316,243; Pgc1α, peroxisome proliferator‐activated receptor‐γ coactivator‐1α; Atgl, adipose triglyceride lipase; Cox2, cytochrome oxidase subunit 2; Cd36, cluster of differentiation 36. C Representative western blot for Ucp1 protein in interscapular BAT and D, densitometric analysis. Data are the mean ± SEM (n = 7‐9 per group). *P < 0.05, **P < 0.01 compared with Control; † P < 0.05, †† P < 0.01 compared with the DM‐Con group. a P = 0.074, b P = 0.072 compared with Control

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  Figure 4. Effects of CL316,243 on serum adipokines (adiponectin and fibroblast growth factor [FGF] 21) and renal expression of Fgf21 components in diabetic mice. A, Serum adiponectin and B, serum Fgf21 concentrations; C, Fgf21 protein and D, relative expression of β‐klotho (Klb) and FGF receptor 1c (Fgfr1c) mRNA in renal cortical tissues. Data are the mean ± SEM (n = 6 per group). *P < 0.05, **P < 0.01, ***P < 0.001 compared with control (non‐diabetic) mice; † P < 0.05, †† P < 0.01 compared with the diabetic control (DM‐Con) group. DM + CL, HFD combined with STZ‐induced diabetic mice treated with CL316,243

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  Figure 5. Effects of CL316,243 on serum microRNAs (miR) and renal expression of target genes in diabetic mice. A, Relative expression of miR‐26a, miR‐30b, miR‐30c, miR‐99a, miR‐100 and miR‐181a extracted from the serum of non‐diabetic (Control), streptozotocin‐induced diabetic (DM‐Con), and CL316,243‐treated diabetic (DM + CL) mice. B, Relative mRNA expression of connective tissue growth factor (Ctgf), Snail1, Nox4, and early growth response factor 1 (Egr1) in renal cortical tissues. Data are the mean ± SEM (n = 6 per group). *P < 0.05, **P < 0.01, ***P < 0.001 compared with Control; † P < 0.05, †† P < 0.01 compared with the DM‐Con group

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  Figure 6. Fibrosis, inflammation, and oxidative stress‐related molecules in the renal cortex of non‐diabetic (Control), streptozotocin‐induced diabetic (DM‐Con), and CL316,243‐treated diabetic (DM + CL) mice. A, Relative expression of the mRNA of renal fibrotic and inflammatory molecules. Tgf‐β1, transforming growth factor‐β1; Col1, collagen 1; Fn, fibronectin; Tnfα, tumor necrosis factor‐α; IL‐1β, interleukin‐1β. B, Representative western blot showing Fn, Col1, Tgf‐β1, Tnf‐α, NADPH oxidase 1 (Nox1), NAD(P)H dehydrogenase 1 (Nqo1), superoxide dismutase 1 (Sod1), and β‐actin. C‐I, Densitometric analysis of western blotting results. Data are the mean ± SEM (n = 6 per group). *P < 0.05, **P < 0.01, ***P < 0.001 compared with Control; † P < 0.05, †† P < 0.01 compared with the DM‐Con group. a P = 0.089 compared with the DM‐Con group

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  Figure 7. Renal histological changes in non‐diabetic (Control), streptozotocin‐induced diabetic (DM‐Con), and CL316,243‐treated diabetic (DM + CL) mice. A,B, Representative images of periodic acid‐Schiff (PAS) and Masson's trichrome (Masson) staining of renal sections from Control, DM‐Con, and DM + CL mice. Bars, 10 μm. C, Mesangial expansion and D, collagen accumulation in PAS‐ and Masson‐stained slices, respectively, as quantified using Image‐Pro Plus 6.0 software (Media Cybernetics, Silver Springs, MD). Data are the mean ± SEM (n = 6 per group). *P < 0.05, **P < 0.01, ***P < 0.001 compared with Control; ††† P < 0.001 compared with the DM‐Con group

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  Figure 8. Phosphorylated (p‐) AMP‐activated protein kinase (AMPK) (Thr172), total AMPK, sirtuin 1 (Sirt1) and peroxisome proliferator‐activated receptor‐γ coactivator‐1α (Pgc1α) levels in the renal cortex of non‐diabetic (Control), streptozotocin‐induced diabetic (DM‐Con), and CL316,243‐treated diabetic (DM + CL) mice. A, Representative western blots for p‐AMPK (Thr172), total AMPK, Sirt1, Pgc1α, and β‐actin in renal cortical tissues. B‐D, Densitometric analysis of western blotting results. Data are the mean ± SEM (n = 6 per group). *P < 0.05, **P < 0.01 compared with Control; † P < 0.05 compared with DM‐Con