Reducing hepatic PKD activity lowers circulating VLDL cholesterol

in Journal of Endocrinology
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  • 1 Institute for Mental and Physical Health and Clinical Translation (iMPACT) and Metabolic Research Unit, School of Medicine, Deakin University, Geelong, Victoria, Australia
  • 2 Institute for Health and Sport (iHeS), Victoria University, Melbourne, Victoria, Australia
  • 3 Baker Heart and Diabetes Institute, Melbourne, Victoria, Australia
  • 4 Institute for Physical Activity and Nutrition (IPAN) and School of Exercise and Nutrition Science and Deakin University, Geelong, Victoria, Australia

Correspondence should be addressed to S L McGee: sean.mcgee@deakin.edu.au
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Protein kinase D (PKD) is emerging as an important kinase regulating energy balance and glucose metabolism; however, whether hepatic PKD activity can be targeted to regulate these processes is currently unclear. In this study, hepatic PKD activity was reduced using adeno-associated virus vectors to express a dominant-negative (DN) version of PKD1, which impairs the action of all three PKD isoforms. In chow-fed mice, hepatic DN PKD expression increased whole-body glucose oxidation, but had only mild effects on glucose and insulin tolerance and no effects on glucose homeostasis following fasting and refeeding. However, circulating VLDL cholesterol was reduced under these conditions and was associated with hepatic fatty acid accumulation, but not lipids involved in lipoprotein synthesis. The limited effects on glucose homeostasis in DN PKD mice was despite reduced expression of gluconeogenic genes under both fasted and refed conditions, and enhanced pyruvate tolerance. The requirement for PKD for gluconeogenic capacity was supported by in vitro studies in cultured FAO hepatoma cells expressing DN PKD, which produced less glucose under basal conditions. Although these pathways are increased in obesity, the expression of DN PKD in the liver of mice fed a high-fat diet had no impact on glucose tolerance, insulin action, pyruvate tolerance or plasma VLDL. Together, these data suggest that PKD signalling in the liver regulates metabolic pathways involved in substrate redistribution under conditions of normal nutrient availability, but not under conditions of overnutrition such as in obesity.

Supplementary Materials

    • Figure S1: Determination of PKD abundance using a recombinant GST-PKD standard curve. Recombinant PKD and lysates used for PKD activity assays were run on SDS-PAGE gels and blotted for total PKD.
    • Figure S2: Whole body metabolism in GFP and DN PKD mice. (A) Energy expenditure; (B) Respiratory exchange ratio; (C) Lipid oxidation; (D) Plasma insulin throughout a glucose tolerance test. Data are mean &#x00B1; SEM, n=6/mice per group and were analysed by two-way ANOVA. # signifies main effect for genotype (p<0.05).
    • Figure S3: E1-alpha subunit of PDH phosphorylation at serine 293(A) and CREB phosphorylation at serine 133 (B) in liver from GFP and DN PKD expressing mice following a 16hr fast or a 4hr re-feeding period. Data are mean &#x00B1; SEM, n=3 biological mice/group.
    • Figure S4: Hepatic DN PKD expression in HFD mice. (A) Body weight; (B) Lean mass; (C) Fat mass; (D) Energy expenditure; (E) Respiratory exchange ratio. Data are mean &#x00B1; SEM, n=7-8/mice per group.

 

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