Advertisement

Diabetologia

, Volume 61, Issue 5, pp 1180–1192 | Cite as

Follicle-stimulating hormone enhances hepatic gluconeogenesis by GRK2-mediated AMPK hyperphosphorylation at Ser485 in mice

  • Xiaoyi Qi
  • Yanjing Guo
  • Yongfeng Song
  • Chunxiao Yu
  • Lifang Zhao
  • Li Fang
  • Dehuan Kong
  • Jiajun Zhao
  • Ling Gao
Article

Abstract

Aims/hypothesis

Increased serum follicle-stimulating hormone (FSH) is correlated with fasting hyperglycaemia. However, the underlying mechanism remains unclear. Because excessive hepatic gluconeogenesis is a major cause of fasting hyperglycaemia the present study investigated whether FSH increases hepatic gluconeogenesis in mice.

Methods

Ovariectomised mice supplemented with oestradiol (E2) to maintain normal levels of serum E2 (OVX+E2 mice) were injected with low or high doses of FSH. We knocked out Crtc2, a crucial factor in gluconeogenesis, and Fshr to discern their involvement in FSH signalling. To evaluate the role of the G-protein-coupled receptor (GPCR) kinase 2 (GRK2), which could affect glucose metabolism and interact directly with non-GPCR components, a specific GRK2 inhibitor was used. The pyruvate tolerance test (PTT), quantification of PEPCK and glucose-6-phosphatase (G6Pase), key enzymes of gluconeogenesis, GRK2 and phosphorylation of AMP-activated protein kinase (AMPK) were examined to evaluate the level of gluconeogenesis in the liver. A nonphosphorylatable mutant of AMPK Ser485 (AMPK S485A) was transfected into HepG2 cells to evaluate the role of AMPK Ser485 phosphorylation.

Results

FSH increased fasting glucose (OVX+E2+high-dose FSH 8.18 ± 0.60 mmol/l vs OVX+E2 6.23 ± 1.33 mmol/l), the PTT results, and the transcription of Pepck (also known as Pck1; 2.0-fold increase) and G6pase (also known as G6pc; 2.5-fold increase) in OVX+E2 mice. FSH also enhanced the promoter luciferase activities of the two enzymes in HepG2 cells. FSH promoted the membrane translocation of GRK2, which is associated with increased AMPK Ser485 and decreased AMPK Thr172 phosphorylation, and enhanced the nuclear translocation of cyclic AMP-regulated transcriptional coactivator 2 (CRTC2). GRK2 could bind with AMPK and induce Ser485 hyperphosphorylation. Furthermore, either the GRK2 inhibitor or AMPK S485A blocked FSH-regulated AMPK Thr172 dephosphorylation and gluconeogenesis. Additionally, the deletion of Crtc2 or Fshr abolished the function of FSH in OVX+E2 mice.

Conclusions/interpretation

The results indicate that FSH enhances CRTC2-mediated gluconeogenesis dependent on AMPK Ser485 phosphorylation via GRK2 in the liver, suggesting an essential role of FSH in the pathogenesis of fasting hyperglycaemia.

Keywords

AMPK CRTC2 FSH Gluconeogenesis GRK2 Liver 

Abbreviations

AICAR

5-Aminoimidazole-4-carboxamide-1-β-d-ribofuranoside

ALT

Alanine aminotransferase

AMPK

AMP-activated protein kinase

AST

Aspartate aminotransferase

CaMKK

Calcium/calmodulin-dependent protein kinase

CRE

Cyclic AMP response element

CREB

Cyclic AMP response element-binding protein

CRTC2

Cyclic AMP-regulated transcriptional coactivator 2

E2

Oestradiol

FSH

Follicle-stimulating hormone

FSHR

Follicle-stimulating hormone receptor

G6Pase

Glucose-6-phosphatase

GPCR

G-protein-coupled receptor

GRK2

GPCR kinase 2

GSK3β

Glycogen synthase kinase 3β

H&E

Haematoxylin and eosin

H-FSH

High-dose FSH

ITT

Insulin tolerance test

Kitt

Insulin sensitivity index

L-FSH

Low-dose FSH

LKB1

Liver kinase B1

MAPK

Mitogen-activated protein kinase

OVX

Ovariectomised

PAS

Periodic acid–Schiff

PKA

Protein kinase A

PTT

Pyruvate tolerance test

Notes

Acknowledgements

We thank Y. Wang (School of Life Sciences, Tsinghua University, China) for providing mice; H. Choi (School of Biological Sciences and Technology, Chonnam National University, Republic of Korea), J. Staňková (Department of Paediatrics, University of Sherbrooke, Canada), F. Mayor (Department of Molecular Biology, Universidad Autónoma de Madrid, Spain) and J. L. Benovic (Department of Biochemistry and Molecular Biology, Thomas Jefferson University, USA) for providing plasmids.

Contribution statement

JZ and LG designed and supervised the project. JZ, LG, CY, YS, YG and XQ designed the experiments. XQ, YG and DK performed the experiments. XQ analysed the data and wrote the manuscript. CY, DK, LF and LZ participated in analysis and interpretation of data. All authors revised the manuscript critically and approved the final version for publication. LG is the guarantor of this work.

Funding

This work was supported by the National Natural Science Foundation (81670796) and the National Key R&D Programme of China (2017YFC1309800 and 0909600).

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2018_4562_MOESM1_ESM.pdf (626 kb)
ESM (PDF 626 kb)

References

  1. 1.
    Heianza Y, Arase Y, Kodama S et al (2013) Effect of postmenopausal status and age at menopause on type 2 diabetes and prediabetes in Japanese individuals: Toranomon Hospital Health Management Center Study 17 (TOPICS 17). Diabetes Care 36:4007–4014CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Mauvais-Jarvis F, Clegg DJ, Hevener AL (2013) The role of estrogens in control of energy balance and glucose homeostasis. Endocr Rev 34:309–338CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Kulaksizoglu M, Ipekci SH, Kebapcilar L et al (2013) Risk factors for diabetes mellitus in women with primary ovarian insufficiency. Biol Trace Elem Res 154:313–320CrossRefPubMedGoogle Scholar
  4. 4.
    Banerjee AA, Mahale SD (2015) Role of the extracellular and intracellular loops of follicle-stimulating hormone receptor in its function. Front Endocrinol 6:110CrossRefGoogle Scholar
  5. 5.
    Liu XM, Chan HC, Ding GL et al (2015) FSH regulates fat accumulation and redistribution in aging through the Galphai/Ca(2+)/CREB pathway. Aging Cell 14:409–420CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Onori P, Mancinelli R, Franchitto A et al (2013) Role of follicle-stimulating hormone on biliary cyst growth in autosomal dominant polycystic kidney disease. Liver Int 33:914–925CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Song Y, Wang ES, Xing LL et al (2016) Follicle-stimulating hormone induces postmenopausal dyslipidemia through inhibiting hepatic cholesterol metabolism. J Clin Endocrinol Metab 101:254–263CrossRefPubMedGoogle Scholar
  8. 8.
    Sun L, Peng Y, Sharrow AC et al (2006) FSH directly regulates bone mass. Cell 125:247–260CrossRefPubMedGoogle Scholar
  9. 9.
    Vila-Bedmar R, Cruces-Sande M, Lucas E et al (2015) Reversal of diet-induced obesity and insulin resistance by inducible genetic ablation of GRK2. Sci Signal 8:ra73CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Taguchi K (2015) The role of GRK2 and its potential as a new therapeutic target in diabetic vascular complications. J Pharm Soc Jpn 135:961–967CrossRefGoogle Scholar
  11. 11.
    Gurevich EV, Tesmer JJ, Mushegian A, Gurevich VV (2012) G protein-coupled receptor kinases: more than just kinases and not only for GPCRs. Pharmacol Ther 133:40–69CrossRefPubMedGoogle Scholar
  12. 12.
    Sharabi K, Tavares CD, Rines AK, Puigserver P (2015) Molecular pathophysiology of hepatic glucose production. Mol Asp Med 46:21–33CrossRefGoogle Scholar
  13. 13.
    Jing Y, Liu W, Cao H et al (2015) Hepatic p38alpha regulates gluconeogenesis by suppressing AMPK. J Hepatol 62:1319–1327CrossRefPubMedGoogle Scholar
  14. 14.
    Rui L (2014) Energy metabolism in the liver. Compr Physiol 4:177–197CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Jitrapakdee S (2012) Transcription factors and coactivators controlling nutrient and hormonal regulation of hepatic gluconeogenesis. Int J Biochem Cell Biol 44:33–45CrossRefPubMedGoogle Scholar
  16. 16.
    Koo SH, Flechner L, Qi L et al (2005) The CREB coactivator TORC2 is a key regulator of fasting glucose metabolism. Nature 437:1109–1111CrossRefPubMedGoogle Scholar
  17. 17.
    Ning J (2011) Suppression of AMPK activation via S485 phosphorylation by IGF-I during hyperglycemia is mediated by AKT activation in vascular smooth muscle cells. Endocrinology 152:3143–3154CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Cao J, Meng S, Chang E et al (2014) Low concentrations of metformin suppress glucose production in hepatocytes through AMP-activated protein kinase (AMPK). J Biol Chem 289:20,435–20,446CrossRefGoogle Scholar
  19. 19.
    Chen Z, Gaudreau R, Le Gouill C, Rola-Pleszczynski M, Stankova J (2004) Agonist-induced internalization of leukotriene B(4) receptor 1 requires G-protein-coupled receptor kinase 2 but not arrestins. Mol Pharmacol 66:377–386CrossRefPubMedGoogle Scholar
  20. 20.
    Kim YD, Park KG, Lee YS et al (2008) Metformin inhibits hepatic gluconeogenesis through AMP-activated protein kinase-dependent regulation of the orphan nuclear receptor SHP. Diabetes 57:306–314CrossRefPubMedGoogle Scholar
  21. 21.
    Miyake K, Ogawa W, Matsumoto M, Nakamura T, Sakaue H, Kasuga M (2002) Hyperinsulinemia, glucose intolerance, and dyslipidemia induced by acute inhibition of phosphoinositide 3-kinase signaling in the liver. J Clin Invest 110:1483–1491CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Tao W, Wu J, Xie BX et al (2015) Lipid-induced muscle insulin resistance is mediated by GGPPS via modulation of the RhoA/Rho kinase signaling pathway. J Biol Chem 290:20,086–20,097CrossRefGoogle Scholar
  23. 23.
    Liu NC, Lin WJ, Kim E et al (2007) Loss of TR4 orphan nuclear receptor reduces phosphoenolpyruvate carboxykinase-mediated gluconeogenesis. Diabetes 56:2901–2909CrossRefPubMedGoogle Scholar
  24. 24.
    Gao D, Nong S, Huang X et al (2010) The effects of palmitate on hepatic insulin resistance are mediated by NADPH oxidase 3-derived reactive oxygen species through JNK and p38MAPK pathways. J Biol Chem 285:29,965–29,973CrossRefGoogle Scholar
  25. 25.
    Malla R, Wang Y, Chan WK, Tiwari AK, Faridi JS (2015) Genetic ablation of PRAS40 improves glucose homeostasis via linking the AKT and mTOR pathways. Biochem Pharmacol 96:65CrossRefPubMedGoogle Scholar
  26. 26.
    Zhang X, Xie X, Heckmann BL, Saarinen AM, Czyzyk TA, Liu J (2014) Targeted disruption of G0/G1 switch gene 2 enhances adipose lipolysis, alters hepatic energy balance, and alleviates high-fat diet-induced liver steatosis. Diabetes 63:934–946CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Gerich JE (2010) Role of the kidney in normal glucose homeostasis and in the hyperglycaemia of diabetes mellitus: therapeutic implications. Diabetic Med 27:136–142CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Lin HV, Accili D (2011) Hormonal regulation of hepatic glucose production in health and disease. Cell Metab 14:9–19CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Rice S, Elia A, Jawad Z, Pellatt L, Mason HD (2013) Metformin inhibits follicle-stimulating hormone (FSH) action in human granulosa cells: relevance to polycystic ovary syndrome. J Clin Endocrinol Metab 98:E1491–E1500CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Chen W, Sang JY, Liu DJ et al (2013) Desensitization of G-protein-coupled receptors induces vascular hypocontractility in response to norepinephrine in the mesenteric arteries of cirrhotic patients and rats. Hepatobiliary Pancreat Dis Int 12:295–304CrossRefPubMedGoogle Scholar
  31. 31.
    Taguchi K, Matsumoto T, Kamata K, Kobayashi T (2012) Inhibitor of G protein-coupled receptor kinase 2 normalizes vascular endothelial function in type 2 diabetic mice by improving beta-arrestin 2 translocation and ameliorating Akt/eNOS signal dysfunction. Endocrinology 153:2985–2996CrossRefPubMedGoogle Scholar
  32. 32.
    Viollet B, Foretz M, Guigas B et al (2006) Activation of AMP-activated protein kinase in the liver: a new strategy for the management of metabolic hepatic disorders. J Physiol 574:41–53CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Park JS, Cho MH, Ahn CW, Kim KR, Huh KB (2012) The association of insulin resistance and carotid atherosclerosis with thigh and calf circumference in patients with type 2 diabetes. Cardiovasc Diabetol 11:62CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Penela P, Murga C, Ribas C, Lafarga V, Mayor F Jr (2010) The complex G protein-coupled receptor kinase 2 (GRK2) interactome unveils new physiopathological targets. Br J Pharmacol 160:821–832CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Penela P, Ribas C, Mayor F Jr (2003) Mechanisms of regulation of the expression and function of G protein-coupled receptor kinases. Cell Signal 15:973–981CrossRefPubMedGoogle Scholar
  36. 36.
    Jurado-Pueyo M, Campos PM, Mayor F, Murga C (2008) GRK2-dependent desensitization downstream of G proteins. J Recept Signal Transduct Res 28:59–70CrossRefPubMedGoogle Scholar
  37. 37.
    Evron T, Daigle TL, Caron MG (2012) GRK2: multiple roles beyond G protein-coupled receptor desensitization. Trends Pharmacol Sci 33:154–164CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Liu S, Premont RT, Kontos CD, Zhu S, Rockey DC (2005) A crucial role for GRK2 in regulation of endothelial cell nitric oxide synthase function in portal hypertension. Nat Med 11:952–958CrossRefPubMedGoogle Scholar
  39. 39.
    Peregrin S, Jurado-Pueyo M, Campos PM et al (2006) Phosphorylation of p38 by GRK2 at the docking groove unveils a novel mechanism for inactivating p38MAPK. Curr Biol 16:2042–2047CrossRefPubMedGoogle Scholar
  40. 40.
    Copps KD, White MF (2012) Regulation of insulin sensitivity by serine/threonine phosphorylation of insulin receptor substrate proteins IRS1 and IRS2. Diabetologia 55:2565–2582CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Viollet B, Guigas B, Sanz GN, Leclerc J, Foretz M, Andreelli F (2012) Cellular and molecular mechanisms of metformin: an overview. Clin Sci 122:253–270CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Horman S, Vertommen D, Heath R et al (2006) Insulin antagonizes ischemia-induced Thr172 phosphorylation of AMP-activated protein kinase alpha-subunits in heart via hierarchical phosphorylation of Ser485/491. J Biol Chem 281:5335–5340CrossRefPubMedGoogle Scholar
  43. 43.
    Woods A, Johnstone SR, Dickerson K et al (2003) LKB1 is the upstream kinase in the AMP-activated protein kinase cascade. Curr Biol 13:2004–2008CrossRefPubMedGoogle Scholar
  44. 44.
    Anderson KA, Means RL, Huang QH et al (1998) Components of a calmodulin-dependent protein kinase cascade. Molecular cloning, functional characterization and cellular localization of Ca2+/calmodulin-dependent protein kinase kinase beta. J Biol Chem 273:31,880–31,889CrossRefGoogle Scholar
  45. 45.
    Hasenour CM, Berglund ED, Wasserman DH (2013) Emerging role of AMP-activated protein kinase in endocrine control of metabolism in the liver. Mol Cell Endocrinol 366:152–162CrossRefPubMedGoogle Scholar
  46. 46.
    Altarejos JY, Montminy M (2011) CREB and the CRTC co-activators: sensors for hormonal and metabolic signals. Nat Rev Mol Cell Biol 12:141–151CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Wang Y, Inoue H, Ravnskjaer K et al (2010) Targeted disruption of the CREB coactivator Crtc2 increases insulin sensitivity. Proc Natl Acad Sci U S A 107:3087–3092CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Law NC, White MF, Hunzicker-Dunn ME (2016) G protein-coupled receptors (GPCRs) that signal via protein kinase A (PKA) cross-talk at insulin receptor substrate 1 (IRS1) to activate the phosphatidylinositol 3-kinase (PI3K)/Akt pathway. J Biol Chem 291:27,160–27,169CrossRefGoogle Scholar
  49. 49.
    Landomiel F, Gallay N, Jegot G et al (2014) Biased signalling in follicle stimulating hormone action. Mol Cell Endocrinol 382:452–459CrossRefPubMedGoogle Scholar
  50. 50.
    Saltiel AR, Kahn CR (2001) Insulin signalling and the regulation of glucose and lipid metabolism. Nature 414:799–806CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Xiaoyi Qi
    • 1
    • 2
    • 3
  • Yanjing Guo
    • 1
    • 2
    • 3
  • Yongfeng Song
    • 1
    • 2
    • 3
  • Chunxiao Yu
    • 1
    • 2
    • 3
  • Lifang Zhao
    • 1
    • 2
    • 3
  • Li Fang
    • 1
    • 2
    • 3
  • Dehuan Kong
    • 4
  • Jiajun Zhao
    • 1
    • 2
    • 3
  • Ling Gao
    • 5
  1. 1.Department of EndocrinologyShandong Provincial Hospital affiliated to Shandong UniversityJinanPeople’s Republic of China
  2. 2.Shandong Provincial Key Laboratory of Endocrinology and Lipid MetabolismJinanPeople’s Republic of China
  3. 3.Institute of Endocrinology and MetabolismShandong Academy of Clinical MedicineJinanPeople’s Republic of China
  4. 4.Department of GeriatricsTai’an City Central HospitalTai’anPeople’s Republic of China
  5. 5.Scientific Centre, Shandong Provincial Hospital affiliated to Shandong UniversityJinanPeople’s Republic of China

Personalised recommendations