PRMT1 promotes hyperglycemia in a FoxO1-dependent manner, affecting glucose metabolism, during hypobaric hypoxia exposure, in rat model
High-altitude (HA) environment causes changes in cellular metabolism among unacclimatized humans. Previous studies have revealed that insulin-dependent activation of protein kinase B (Akt) regulates metabolic processes via discrete transcriptional effectors. Moreover, protein arginine methyltransferase (PRMT)1-dependent arginine modification of forkhead box other (FoxO)1 protein interferes with Akt-dependent phosphorylation. The present study was undertaken to test the involvement of PRMT1 on FoxO1 activation during hypobaric hypoxia (HH) exposure in rat model.
Samples were obtained from normoxia control (NC) and HH-exposed (H) rats, subdivided according to the duration of HH exposure. To explore the specific role played by PRMT1 during HH exposure, samples from 1d pair-fed (PF) NC, 1d acute hypoxia-exposed (AH) placebo-treated, and 1d AH TC-E-5003-treated rats were investigated. Quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) was performed to determine expressions of glycolytic, gluconeogenic enzymes, and insulin response regulating genes. Immuno-blot and enzyme linked immunosorbent assay (ELISA) were used for insulin response regulating proteins. Nuclear translocation of FoxO1 was analyzed using deoxyribonucleic acid (DNA)-binding ELISA kit.
We observed HH-induced increase in glycolytic enzyme expressions in hepatic tissue unlike hypothalamic tissue. PRMT1 expression increased during HH exposure, causing insulin resistance and resulting increase in FoxO1 nuclear translocation, leading to hyperglycemia. Conversely, PRMT1 inhibitor treatment promoted inhibition of FoxO1 activity and increase in glucose uptake during HH exposure leading to reduction in blood-glucose and hepatic glycogen levels.
PRMT1 might have a potential importance as a therapeutic target for the treatment of HH-induced maladies.
KeywordsHypobaric hypoxia Insulin resistance Hyperglycemia Protein arginine methyltransferase 1 Protein kinase B Fork head box protein
We thank Dr. N.K. Sethy and Dr. K. Ray for excellent technical support.
A.J.D., P.G., S.B., S.S. were involved in acquisition of data. S.B. conceived and designed the study, analyzed and interpretated the data, and drafted the article also. All authors contributed to the revising of the manuscript, and approved the final version of the manuscript.
The work was funded by the Defence Research and Development Organization (DRDO), Government of India. S.B., D.A.J., and P.G. are thankful to DRDO, and S.S. is thankful to the University Grants Commission (UGC) for junior/senior research fellowships.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
The study protocol was approved by the Institutional Animal Ethics Committee.
- 1.N. Simler, A. Malgoyre, N. Koulmann, A. Alonso, A. Peinnequin, A.X. Bigard, Hypoxic stimulus alters hypothalamic AMP-activated protein kinase phosphorylation concomitant to hypophagia. J. Appl. Physiol. 102(6), 2135–2141 (2007). https://doi.org/10.1152/japplphysiol.01150.2006 CrossRefPubMedGoogle Scholar
- 6.C.M. Taniguchi, T. Kondo, M. Sajan, J. Luo, R. Bronson, T. Asano, R. Farese, L.C. Cantley, C.R. Kahn, Divergent regulation of hepatic glucose and lipid metabolism by phosphoinositide 3-kinase via Akt and PKClambda/zeta. Cell Metab. 3(5), 343–353 (2006). https://doi.org/10.1016/j.cmet.2006.04.005 CrossRefPubMedGoogle Scholar
- 10.H. Ren, I.J. Orozco, Y. Su, S. Suyama, R. Gutierrez-Juarez, T.L. Horvath, S.L. Wardlaw, L. Plum, O. Arancio, D. Accili, FoxO1 target Gpr17 activates AgRP neurons to regulate food intake. Cell 149(6), 1314–1326 (2012). https://doi.org/10.1016/j.cell.2012.04.032 CrossRefPubMedPubMedCentralGoogle Scholar
- 12.W. Zhang, S. Patil, B. Chauhan, S. Guo, D.R. Powell, J. Le, A. Klotsas, R. Matika, X. Xiao, R. Franks, K.A. Heidenreich, M.P. Sajan, R.V. Farese, D.B. Stolz, P. Tso, S.H. Koo, M. Montminy, T.G. Unterman, FoxO1 regulates multiple metabolic pathways in the liver: effects on gluconeogenic, glycolytic, and lipogenic gene expression. J. Biol. Chem. 281(15), 10105–10117 (2006). https://doi.org/10.1074/jbc.M600272200 CrossRefPubMedGoogle Scholar
- 15.M. Kobayashi, O. Kikuchi, T. Sasaki, H.J. Kim, H. Yokota-Hashimoto, Y.S. Lee, K. Amano, T. Kitazumi, V.Y. Susanti, Y.I. Kitamura, T. Kitamura, FoxO1 as a double-edged sword in the pancreas: analysis of pancreas- and beta-cell-specific FoxO1 knockout mice. Am. J. Physiol. Endocrinol. Metab. 302(5), E603–E613 (2012). https://doi.org/10.1152/ajpendo.00469.2011 CrossRefPubMedGoogle Scholar
- 17.K. Yamagata, H. Daitoku, Y. Takahashi, K. Namiki, K. Hisatake, K. Kako, H. Mukai, Y. Kasuya, A. Fukamizu, Arginine methylation of FOXO transcription factors inhibits their phosphorylation by Akt. Mol. Cell 32(2), 221–231 (2008). https://doi.org/10.1016/j.molcel.2008.09.013 CrossRefPubMedGoogle Scholar
- 20.K.A. Sikaris, The clinical biochemistry of obesity. The clinical biochemist. Reviews 25(3), 165–181 (2004)Google Scholar
- 22.C.M. Taniguchi, E.C. Finger, A.J. Krieg, C. Wu, A.N. Diep, E.L. LaGory, K. Wei, L.M. McGinnis, J. Yuan, C.J. Kuo, A.J. Giaccia, Cross-talk between hypoxia and insulin signaling through Phd3 regulates hepatic glucose and lipid metabolism and ameliorates diabetes. Nat. Med. 19(10), 1325–1330 (2013). https://doi.org/10.1038/nm.3294 CrossRefPubMedPubMedCentralGoogle Scholar
- 23.K. Wei, S.M. Piecewicz, L.M. McGinnis, C.M. Taniguchi, S.J. Wiegand, K. Anderson, C.W. Chan, K.X. Mulligan, D. Kuo, J. Yuan, M. Vallon, L.C. Morton, E. Lefai, M.C. Simon, J.J. Maher, G. Mithieux, F. Rajas, J.P. Annes, O.P. McGuinness, G. Thurston, A.J. Giaccia, C.J. Kuo, A liver HIF-2alpha-Irs2 pathway sensitizes hepatic insulin signaling and is modulated by Vegf inhibition. Nat. Med. 19(10), 1331–1337 (2013). https://doi.org/10.1038/nm.3295 CrossRefPubMedPubMedCentralGoogle Scholar
- 25.B. Faubert, G. Boily, S. Izreig, T. Griss, B. Samborska, Z. Dong, F. Dupuy, C. Chambers, B.J. Fuerth, B. Viollet, O.A. Mamer, D. Avizonis, R.J. DeBerardinis, P.M. Siegel, R.G. Jones, AMPK is a negative regulator of the Warburg effect and suppresses tumor growth in vivo. Cell Metab. 17(1), 113–124 (2013). https://doi.org/10.1016/j.cmet.2012.12.001 CrossRefPubMedGoogle Scholar
- 26.S.P. Mathupala, A. Rempel, P.L. Pedersen, Glucose catabolism in cancer cells: identification and characterization of a marked activation response of the type II hexokinase gene to hypoxic conditions. J. Biol. Chem. 276(46), 43407–43412 (2001). https://doi.org/10.1074/jbc.M108181200 CrossRefPubMedGoogle Scholar
- 27.U. Roth, K. Curth, T.G. Unterman, T. Kietzmann, The transcription factors HIF-1 and HNF-4 and the coactivator p300 are involved in insulin-regulated glucokinase gene expression via the phosphatidylinositol 3-kinase/protein kinase B pathway. J. Biol. Chem. 279(4), 2623–2631 (2004). https://doi.org/10.1074/jbc.M308391200 CrossRefPubMedGoogle Scholar
- 30.A. Minchenko, I. Leshchinsky, I. Opentanova, N. Sang, V. Srinivas, V. Armstead, J. Caro, Hypoxia-inducible factor-1-mediated expression of the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 (PFKFB3) gene. Its possible role in the Warburg effect. J. Biol. Chem. 277(8), 6183–6187 (2002). https://doi.org/10.1074/jbc.M110978200 CrossRefPubMedGoogle Scholar
- 31.S. Guo, G. Rena, S. Cichy, X. He, P. Cohen, T. Unterman, Phosphorylation of serine 256 by protein kinase B disrupts transactivation by FKHR and mediates effects of insulin on insulin-like growth factor-binding protein-1 promoter activity through a conserved insulin response sequence. J. Biol. Chem. 274(24), 17184–17192 (1999)CrossRefPubMedGoogle Scholar
- 32.T. Porstmann, B. Griffiths, Y.L. Chung, O. Delpuech, J.R. Griffiths, J. Downward, A. Schulze, PKB/Akt induces transcription of enzymes involved in cholesterol and fatty acid biosynthesis via activation of SREBP. Oncogene 24(43), 6465–6481 (2005). https://doi.org/10.1038/sj.onc.1208802 CrossRefPubMedGoogle Scholar
- 34.M. Milkiewicz, E. Roudier, J.L. Doyle, A. Trifonova, O. Birot, T.L. Haas, Identification of a mechanism underlying regulation of the anti-angiogenic forkhead transcription factor FoxO1 in cultured endothelial cells and ischemic muscle. Am. J. Pathol. 178(2), 935–944 (2011). https://doi.org/10.1016/j.ajpath.2010.10.042 CrossRefPubMedPubMedCentralGoogle Scholar
- 35.S. Zhang, Y. Zhao, M. Xu, L. Yu, Y. Zhao, J. Chen, Y. Yuan, Q. Zheng, X. Niu, FoxO3a modulates hypoxia stress induced oxidative stress and apoptosis in cardiac microvascular endothelial cells. PLoS ONE 8(11), e80342 (2013). https://doi.org/10.1371/journal.pone.0080342 CrossRefPubMedPubMedCentralGoogle Scholar
- 37.S.K. Lim, Y.W. Jeong, D.I. Kim, M.J. Park, J.H. Choi, S.U. Kim, S.S. Kang, H.J. Han, S.H. Park, Activation of PRMT1 and PRMT5 mediates hypoxia- and ischemia-induced apoptosis in human lung epithelial cells and the lung of miniature pigs: the role of p38 and JNK mitogen-activated protein kinases. Biochem. Biophys. Res. Commun. 440(4), 707–713 (2013). https://doi.org/10.1016/j.bbrc.2013.09.136 CrossRefPubMedGoogle Scholar
- 38.K. Hirota, J. Sakamaki, J. Ishida, Y. Shimamoto, S. Nishihara, N. Kodama, K. Ohta, M. Yamamoto, K. Tanimoto, A. Fukamizu, A combination of HNF-4 and Foxo1 is required for reciprocal transcriptional regulation of glucokinase and glucose-6-phosphatase genes in response to fasting and feeding. J. Biol. Chem. 283(47), 32432–32441 (2008). https://doi.org/10.1074/jbc.M806179200 CrossRefPubMedGoogle Scholar
- 39.D.T. Duong, M.E. Waltner-Law, R. Sears, L. Sealy, D.K. Granner, Insulin inhibits hepatocellular glucose production by utilizing liver-enriched transcriptional inhibitory protein to disrupt the association of CREB-binding protein and RNA polymerase II with the phosphoenol pyruvate carboxykinase gene promoter. J. Biol. Chem. 277(35), 32234–32242 (2002). https://doi.org/10.1074/jbc.M204873200 CrossRefPubMedGoogle Scholar
- 42.H. Yamashita, M. Takenoshita, M. Sakurai, R.K. Bruick, W.J. Henzel, W. Shillinglaw, D. Arnot, K. Uyeda, A glucose-responsive transcription factor that regulates carbohydrate metabolism in the liver. Proc. Natl. Acad. Sci. USA 98(16), 9116–9121 (2001). https://doi.org/10.1073/pnas.161284298 CrossRefPubMedPubMedCentralGoogle Scholar
- 44.E.M. Bissinger, R. Heinke, A. Spannhoff, A. Eberlin, E. Metzger, V. Cura, P. Hassenboehler, J. Cavarelli, R. Schule, M.T. Bedford, W. Sippl, M. Jung, Acyl derivatives of p-aminosulfonamides and dapsone as new inhibitors of the arginine methyltransferase hPRMT1. Bioorg. Med. Chem. 19(12), 3717–3731 (2011). https://doi.org/10.1016/j.bmc.2011.02.032 CrossRefPubMedGoogle Scholar