Disruption of endothelial Pfkfb3 ameliorates diet-induced murine insulin resistance

in Journal of Endocrinology
Authors:
Qiuhua Yang Vascular Biology Center, Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, USA

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Jiean Xu Vascular Biology Center, Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, USA

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Qian Ma Vascular Biology Center, Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, USA

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Zhiping Liu Vascular Biology Center, Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, USA

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Yaqi Zhou State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, Peking University, Shenzhen, China

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Yongfeng Cai State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, Peking University, Shenzhen, China

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Xiaoxiao Mao Vascular Biology Center, Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, USA

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David Stepp Vascular Biology Center, Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, USA

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Neal Weintraub Vascular Biology Center, Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, USA

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David J Fulton Vascular Biology Center, Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, USA

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Mei Hong State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, Peking University, Shenzhen, China

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Yuqing Huo Vascular Biology Center, Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, USA

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Correspondence should be addressed to Y Huo: YHUO@augusta.edu
DOI:
https://doi.org/10.1530/JOE-20-0524
Volume/Issue:
Volume 250: Issue 3
Article Type:
Research Article
Page Range:
93–104
Online Publication Date:
14 Jul 2021
Copyright:
© Society for Endocrinology 2021
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Overnutrition-induced endothelial inflammation plays a crucial role in high-fat diet (HFD)-induced insulin resistance in animals. Endothelial glycolysis plays a critical role in endothelial inflammation and proliferation, but its role in diet-induced endothelial inflammation and subsequent insulin resistance has not been elucidated. PFKFB3 is a critical glycolytic regulator, and its increased expression has been observed in adipose vascular endothelium of C57BL/6J mice fed with HFD in vivo, and in palmitate (PA)-treated primary human adipose microvascular endothelial cells (HAMECs) in vitro. We generated mice with Pfkfb3 deficiency selective for endothelial cells to examine the effect of endothelial Pfkfb3 in endothelial inflammation in metabolic organs and in the development of HFD-induced insulin resistance. EC Pfkfb3-deficientmice exhibited mitigated HFD-induced insulin resistance, including decreased body weight and fat mass, improved glucose clearance and insulin sensitivity, and alleviated adiposity and hepatic steatosis. Mechanistically, cultured PFKFB3 knockdown HAMECs showed decreased NF-κB activation induced by PA, and consequent suppressed adhesion molecule expression and monocyte adhesion. Taken together, these results demonstrate that increased endothelial PFKFB3 expression promotes diet-induced inflammatory responses and subsequent insulin resistance, suggesting that endothelial metabolic alteration plays an important role in the development of insulin resistance.

Supplementary Materials

    • Supplementary Figure 1. A. Body weight of Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD at the age of 16 weeks. n = 5. B. Fat and lean content of Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD. n = 5. C. Fasting glucose and insulin of Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD. D. Blood glucose levels (left) and AUC (area under the curve, right) during GTT (glucose tolerance test) in Pfkfb3WT and Pfkfb3ΔVEC mice fed with CD. n = 5. Mice were fasted for 6 h and injected with glucose (1 g/kg i.p.). E. Blood glucose levels (left) and AUC (right) during ITT (insulin tolerance test) in Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD. n = 5. Mice were fasted for 4 h and injected with insulin (1 unit/kg body weight through intraperitoneal injection). n = 5. F. Representative images of hematoxylin and eosin (H&E) and Mac2 staining of epididymal WAT sections from Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD. n = 5 mice / group, 4 areas / mice were quantified. G. Representative images of hematoxylin and eosin (H&E) and Mac2 staining of liver sections from Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD. n = 5 mice / group, 4 areas / mice were quantified. All data were expressed as mean ± SEM. Statistical significance was determined by unpaired Student’s t-test. * p < 0.05 was considered significant, ** p < 0.01, *** p < 0.001.
    • Supplementary Figure 2. A. Oxygen consumption (VO2) of Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD during 12-h light and dark cycles recorded at the second day after acclimatization, n = 4. B. Carbon dioxide consumption (VCO2) of Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD during 12-h light and dark cycles recorded at the second day after acclimatization, n = 4. C. Respiratory exchange ratio (RER) of Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD during 12-h light and dark cycles recorded at the second day after acclimatization, n = 4. D. Daily food intake and drink of Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD during light and dark cycles of animals kept on CD at room temperature (22°C), n = 4. E. Heat production of Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD during 12-h light and dark cycles, n = 4. F. Locomotor activity of Pfkfb3WT and Pfkfb3ΔVEC male mice fed with CD during 12-h light and dark cycles, n = 4. Area under the curve (AUC) was calculated during light and dark cycles for each individual animal. Black horizontal bars denote the dark period of the day (12 h). AUC, area under the curve. All data were expressed as mean ± SEM. Statistical significance was determined by unpaired Student’s t-test. *p < 0.05 was considered significant, **p < 0.01, ***p < 0.001.
    • Supplementary Figure 3. A. Schematic diagram of endothelial-specific Pfkfb3-deficient mouse generation. We crossed Pfkfb3flox/flox mice with Cdh5cre transgenic mice to generate endothelial-specific Pfkfb3-deficient mice. B. Schematic diagram of HFD-induced insulin resistance model. Pfkfb3WT and Pfkfb3ΔVEC mice were fed with HFD for 12 weeks beginning at the age of 6 weeks, body weight was measured weekly, and metabolic parameters were recorded with CLAMS system at 10 weeks of HFD, followed by glucose tolerance test (GTT), insulin tolerance test (ITT) and nuclear magnetic resonance (NMR) at 11 or 12 weeks of HFD. Mice were euthanized after 12 weeks of HFD and major metabolic organs or tissues were harvested and processed for further analyses.

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Print ISSN:
0022-0795
Online ISSN:
1479-6805
Copyright:
© Society for Endocrinology 2021
Page(s):
12
Article Type:
Research Article
Received Date:
12 May 2021
Accepted Date:
16 May 2021
Print Publication Date:
01 Sep 2021
Online Publication Date:
14 Jul 2021
Keywords:
endothelial cell; insulin resistance; PFKFB3; NF-κB; inflammation

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