Dysregulation of mitochondrial dynamics mediated aortic perivascular adipose tissue-associated vascular reactivity impairment under excessive fructose intake

Figures

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  Fig. 1. Coenzyme Q10 treatment reversed the HFD-induced suppression of adipose browning factors in the aortic PVAT. A Schematic illustration of the study design to evaluate high fructose diet (HFD)-induced metabolic changes. The special diet was started at 8 weeks old for 3 months. The metabolic indices were measured before and at the end of the special diet at 8 and 20 weeks of age. Coenzyme Q10 was applied to drinking water (10 mg/100 mL) for 4 weeks from 16 to 20 weeks of age. Then, B plasma adiponectin, C representative gel and densitometric analyses of intracellular UCP1, (D) the representative images of Hematoxylin & Eosin (H&E) staining, and E representative images of UCP1 (red) in aortic PVAT at the age of 20 weeks. Values are the mean ± SEM, n = 14 for each group. *P < 0.05 versus the ND group and ##P < 0.01 versus the HFD group in the post hoc Tukey’s multiple range test. ND: regular diet, HFD, high fructose diet, HFD + Q, HFD with coenzyme Q10. DAPI, 4′,6-diamidino-2-phenylindole, a blue-fluorescent nuclear stain. PVAT, aortic perivascular adipose tissue, UCP, uncoupling protein, β-actin as the internal control of Western blot analyses. Scale bars: 100 μm (Fig. 1D, upper…

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  Fig. 2. Coenzyme Q10 reversed the HFD-reduced mitochondrial mass and respiratory complexes in aortic PVAT. A Mitochondrial mass (green, stained by MitoTracker) and representative gel and densitometric analyses of B mt CPX1, C mt CPX2, D mt CPX3, E mt CPX4-1, F mt CPX4-2, and G mt CPX5 in the aortic PVAT of the ND, HFD and HFD + Q groups at the age of 20 weeks. Values are the mean ± SEM, n = 14 for each group. *P < 0.05, **P < 0.01, ***P < 0.001 versus the ND group and #P < 0.05, ##P < 0.01, ###P < 0.001 versus the HFD group in the post hoc Tukey’s multiple range test. ND, normal diet, HFD, high fructose diet, HFD + Q, HFD with coenzyme Q10. mt CPX, mitochondrial respiratory complex subunit, β-actin as the internal control of Western blot analyses. Scale bars: 10 μm

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  Fig. 3. Coenzyme Q10 reversed the HFD-downregulated mitochondrial biogenesis regulators in aortic PVAT. A The ATP content, representative gel and densitometric analyses of B PGC-1α and C TFAM as well as D mtDNA copy number in the aortic PVAT of the ND, HFD, and HFD + Q groups at the age of 20 weeks. Values are the mean ± SEM, n = 14 for each group. *P < 0.05, **P < 0.01 versus the ND group, and #P < 0.05 versus the HFD group in the post hoc Tukey’s multiple range test. ND: normal diet, HFD: high fructose diet, HFD + Q: HFD with coenzyme Q10. β-actin was used as the internal control for Western blot analyses. PGC-1α: peroxisome proliferator-activated receptor γ coactivator 1 α, TFAM: mitochondrial transcription factor A, mtDNA: mitochondrial DNA, 18S ribosomal RNA (18S) as the internal control of QPCR

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  Fig. 4. Coenzyme Q10 reversed the HFD-induced suppression of mitochondrial fusion in aortic PVAT. Representative gel and densitometric analyses of A OPA1, B MFN1, and C MFN2 in the aortic PVAT of the ND, HFD and HFD + Q groups at the age of 20 weeks. Values are the mean ± SEM, n = 9 for each group. *P < 0.05, **P < 0.01 versus the ND group and #P < 0.05 versus the HFD group in the post hoc Tukey’s multiple range test. ND, normal diet, HFD, high fructose diet, HFD + Q, HFD with coenzyme Q10. OPA1, mitochondrial Dynamin-Like GTPase, MFN, mitofusin, β-actin as the internal control

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  Fig. 5. Coenzyme Q10 reversed HFD-induced mitochondrial fission in aortic PVAT. Representative gel and densitometric analyses of A FIS, B DRP1, C p-DRP1 and D the ratio of p-DRP1/DPR1 in the aortic PVAT of the ND, HFD and HFD + Q groups at the age of 20 weeks. Values are the mean ± SEM, n = 8 for each group. *P < 0.05, **P < 0.01, ***P < 0.001 versus the ND group and #P < 0.05 versus the HFD group in the post hoc Tukey’s multiple range test. ND, normal diet, HFD, high fructose diet, HFD + Q, HFD with coenzyme Q10. FIS1, mitochondrial fission 1 protein, DRP1, dynamin-related protein 1, p-DRP1, phospho-DRP1, β-actin as the internal control

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  Fig. 6. Coenzyme Q10 treatment reversed the HFD-induced increase in fibrotic and calcification proteins in aortic PVAT. Representative gel and densitometric analyses of A α-SMA and B osteocalcin in the aortic PVAT of the ND, HFD and HFD + Q groups at the age of 20 weeks. Values are the mean ± SEM, n = 4 for each group. *P < 0.05, **P < 0.01 versus the ND group and ##P < 0.01, ###P < 0.001 versus the HFD group in the post hoc Tukey’s multiple range test. ND, normal diet, HFD, high fructose diet, HFD + Q, HFD with coenzyme Q10. α-SMA, α-smooth muscle actin, β-actin as the internal control

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  Fig. 7. Coenzyme Q10 treatment reversed the HFD-impaired vascular reactivity of the thoracic aorta. Vascular responses to A KCl, B PE, or C Ach of the aorta in the ND, HFD and HFD + Q groups at the age of 20 weeks. Values are the mean ± SEM, n = 7 for each group. *P < 0.05, **P < 0.01, ***P < 0.001 versus the ND group and #P < 0.05, ##P < 0.01 versus the HFD group in the post hoc Tukey’s multiple range test. ND, normal diet, HFD, high fructose diet, HFD + Q, HFD with coenzyme Q10. PE, phenylephrine, Ach, acetylcholine