Insulin Inhibits the Expression of Adiponectin and AdipoR2 mRNA in Cultured Bovine Adipocytes

  • Sun, Y.G. (College of Veterinary Medicine, Northwest A& F University) ;
  • Zan, L.S. (College of Animal Science and Technology, Northwest A& F University) ;
  • Wang, H.B. (College of Animal Science and Technology, Northwest A& F University) ;
  • Guo, H.F. (College of Veterinary Medicine, Northwest A& F University) ;
  • Yang, D.P. (College of Animal Science and Technology, Northwest A& F University) ;
  • Zhao, X.L. (College of Animal Science and Technology, Northwest A& F University) ;
  • Gui, L.S. (College of Animal Science and Technology, Northwest A& F University)
  • Received : 2009.03.07
  • Accepted : 2009.06.04
  • Published : 2009.10.01


Adiponectin is an adipocyte-derived protein that has a regulatory role in energy homeostasis and influences insulin sensitivity. Its effects on glucose utilization and lipid metabolism are mediated by AdipoR1 and AdipoR2. How insulin affects adiponectin gene expression and secretion is still controversial. This study was conducted to determine the expression of adiponectin, AdipRs and $PPAR-\gamma$ during the differentiation of bovine preadipocytes and the effect of insulin on expression of these genes in bovine adipocytes. The bovine preadipocytes started to accumulate lipids three days after differentiation was induced, with increased expression of adiponectin, AdipoR2 and $PPAR-\gamma$ mRNAs. Insulin decreased the expression of adiponectin mRNA in a dose- and time-dependent fashion, and the inhibition was detectable at insulin concentrations as low as 10 nM and as early as 2 h after addition of 100 nM insulin. Insulin also inhibited the expression of AdipoR2 mRNA at concentrations from 1 to 1,000 nM or 24 h after addition of 100 nM insulin, but did not affect the expression of AdipoR1 in bovine adipocytes. Inhibition of PI3K with LY294002 reversed the inhibition of adiponectin and AdipoR2 mRNA expression by insulin. These results suggest that insulin suppresses the expression of adiponectin and AdipoR2 at least partially via the PI3K signal pathway.


  1. Bodles, A., A. Banga, N. Rasouli, F. Ono, P. A. Kern and R. J. Owens. 2006, Pioglitazone increases secretion of high molecular weight adiponectin from adipocytes, Am. J. Physiol. Endocrinol. Metab 291:E1100-E1105
  2. Fernyhough, M. E., E. Okine, G. Hausman, J. L. Vierck and M. V. Dodson. 2007, PPAR-$\alpha$ and GLUT-4 expression as developmental regulators/markers for preadipocyte differentiation into an adipocyte, Domest. Anim. Endocrinol33:367-378
  3. Fruebis, J., T. S. Tsao, S. Javorschi, D. Ebbets-Reed, M. R. Erickson and F. T. Yen. 2001, Proteolytic cleavage product of 30-kDa adipocyte complement-related protein increases fatty acid oxidation in muscle and causes weight loss in mice, Proc. Natl. Acad. Sci. USA. 98:2005-2010
  4. Maeda, K., K. Okubo, I. Shimomura, T. Funahashi, Y. Matsuzawa and K. Matsubara. 1996, cDNA cloning and expression of a novel adipose specific collagen-like factor, apM1 (AdiPose Most abundant Gene transcript 1), Biochem. Biophys. Res. Commun 221:286-289
  5. Maeda, N., M. Takahashi, T. Funahashi, S. Kihara, H. Nishizawa, K. Kishida, H. Nagaretani, M. Matsuda, R. Komuro, N. Ouchi, H. Kuriyama, K. Hotta, T. Nakamura, I. Shimomura and Y. Matsuzawa. 2001, PPAR gamma ligands increase expression and plasma concentrations of adiponectin, an adipose-derived protein, Diabetes 50:2094-2099
  6. Virkamaki, A., K. Ueki and C. R. Kahn. 1999, Protein-protein interaction in insulin signaling and the molecular mechanisms of insulin resistance, J. Clin. Invest. 103:931-943
  7. Wang, C., X. Mao, L. Wang, M. Liu, M. D. Wetzel and K. L. Guan. 2007, Adiponectin sensitizes insulin signaling by reducing p70S6 kinasemediated serine phosphorylation of IRS-1, J. Biol. Chem 282:7991-7996
  8. Chinetti, G., C. Zawadski, J. C. Fruchart and B. Staels. 2004, Expression of adiponectin receptors in human macrophages and regulation by agonists of the nuclear receptors PPARalpha, PPARgamma, and LXR, Biochem. Biophys. Res. Commun314:151-158
  9. Scherer, P. E., S. Williams, M. Fogliano, G. Baldini and H. F. Lodish. 1995, A novel serum protein similar to C1q, produced exclusively in adipocytes, J. Biol. Chem 270:26746-26749
  10. Yamauchi, T., J. Kamon, H. Waki, Y. Imai, N. Shimozawa, K. Hioki, S. Uchida, Y. Ito, K. Takakuwa, J. Matsui, M. Takata, K. Eto, Y. Terauchi, K. Komeda, M. Tsunoda, K. Murakami, Y. Ohnishi, T. Naitoh, K. Yamamura, Y. Ueyama, P. Froguel, S. Kimura, R. Nagai and T. Kadowaki. 2003, Globular adiponectin protected ob/ob mice from diabetes and ApoEdeficient mice from atherosclerosis, J. Biol. Chem 278:2461- 2468
  11. Fasshauer, M., S. Kralisch, M. Klier, U. Lossner, M. Bluher and J. Klein. 2003, Adiponectin gene expression and secretion is inhibited by interleukin-6 in 3T3-L1 adipocyte, Biochem. Biophys. Res. Commun 301:1045-1050
  12. Ritchie, H. D., S. R. Rust, A. Merkel and W. G. Bergen. 1993, Getting rid of excess fat is not an easy task, Feedstuffs 13:279
  13. Bauche, I. B., E. Ait, S. Mkadem, R. Rezsohazy, T. Funahashi, N. Maeda and L. M. Miranda. 2006, Adiponectin downregulates its own production and the expression of its AdipoR2 in transgenic mice, Biochem. Biophys. Res. Commun 345:1414-1424
  14. Fasshauer, M., J. Klein, S. Neumann, M. Eszlinger and R. Paschke. 2002, Hormonal regulation of adiponectin gene expression in 3T3-L1 adipocytes, Biochem. Biophys. Res. Commun290:1084-1089
  15. Seo, J. B., H. M. Moon, M. J. Noh, Y. S. Lee, H. W. Jeong and E. J. Yoo. 2004, Adipocyte determination-and differentiationdependent factor 1/sterol regulatory element-binding protein 1c regulates mouse adiponectin expression, J. Biol. Chem 279:22108-22117
  16. Yamauchi, T., J. Kamon, H. Waki, Y. Terauchi, N. Kubota and K. Hara. 2001, The fat-derived hormone adiponectin reverses insulin resistance associated with both lipoatrophy and obesity,Nat. Med 7:941-946
  17. Yamauchi, T., J. Kamon, Y. Ito, A. Tsuchida, T. Yokomizo and S. Kita. 2003, Cloning of adiponectin receptors that mediate antidiabetic metabolic effects, Nature 423:762-769
  18. Fu, Y., N. Luo, R. L. Klein and W. T. Garvey. 2005, Adiponectin promotes adipocyte differentiation, insulin sensitivity, and lipid accumulation, J. Lipid Res 46:1369-1379
  19. Sato, C., Z. Yasukawa, N. Honda, T. Matsuda and K. Kitajima. 2001, Identification and adipocyte differentiation-dependent expression of the unique disialic acid residue in an adipose tissue-specific glycoprotein, adipoQ. J. Biol Chem276:28849-28856
  20. Liu, B. H., Y. C. Wang, S. C. Wu, H. J. Mersmanna, W. T. Cheng and S. T. Ding. 2008, Insulin regulates the expression of adiponectin and adiponectin receptors in porcine adipocytes, Domest. Anim. Endocrinol 34:352-359
  21. Nakano, Y., T. Tobe, N. H. Choi-Miura, T. Mazda and M. Tomita. 1996, Isolation and characterization of GBP28, a novel gelatin binding protein, J. Biol. Chem. 120:803-812
  22. Saltiel, A. R. and C. R. Kahn. 2001, Insulin signalling and the regulation of glucose and lipid metabolism, Nature 414:799-806
  23. Ohyama, M., K. Matsuda, S. Torii, T. Matsui, H. Yano and T. Kawada. 1998, The interaction between vitamin A and thiazolidinedion on bovine adipocyte differentiation in primary culture, J. Anim. Sci 76:61-65
  24. Combs, T. P., A. H. Berg, S. Obici, P. E. Scherer and L. Rossetti. 2001, Endogenous glucose production is inhibited by the adiposederived protein Acrp30, J. Clin. Invest 108:1875-1881
  25. Bluher, M., M. D. Michael, O. D. Peroni, K. Ueki, N. Carter and B. B. Kahn. 2002, Adipose tissue selective insulin receptor knockout protects against obesity and obesity-related glucose intolerance, Dev. Cell 3:25-38
  26. Hu, E., P. Liang and B. M. Spiegelman. 1999, AdipoQ is a novel adipose-specific gene dysregulated in obesity, J. Biol. Chem 271:10697-10703
  27. Pajvani, U. B., Hawkins, M., Combs, T. P., Rajala, M. W., Doebber, T. and Berger, J. P. 2004, Complex distribution, not absolute amount of adiponectin, correlates with thiazolidinedionemediated improvement in insulin sensitivity, J. Biol. Chem279:12152-12162
  28. Kovacic, S., C. L. Soltys, A. J. Barr, I. Shiojima, K. Walsh and J. R. Dyck. 2003, Akt activity negatively regulates phosphorylation of AMP-activated protein kinase in the heart, J. Biol. Chem 278:39422-39427
  29. Halleux, C. M., M. Takahashi, M. L. Delporte, R. Detry, T. Funahashi and Y. Matsuzawa. 2001, Secretion of adiponectin and regulation of apM1 gene expression in human visceral adipose tissue, Biochem. Biophys. Res Commun 288:1102-1107
  30. Kadowaki, T., T. Yamauchi, N. Kubota, K. Hara, K. Ueki and K. Tobe. 2006, Adiponectin and adiponectin receptors in insulin resistance, diabetes, and the metabolic syndrome, J. Clin. Invest 116:1784-1792
  31. Pajvani, U. B., X. Du, T. P. Combs, A. H. Berg, M. W. Rajala and T. Schulthess. 2003, Structure–function studies of the adipocyte-secreted hormone acrp30/adiponectin. Implications for metabolic regulation and bioactivity, J. Biol. Chem.278:9073–9085
  32. Yamauchi, T., J. Kamon, Y. Minokoshi, Y. Ito, H. Waki and S. Uchida. 2002, Adiponectin stimulates glucose utilization and fatty acid oxidation by activating AMP-activated protein kinase. Nat. Med 8:1288-1295
  33. Mohamed, S., K. Kazuhiro, A. Mohamed, Y. Daisuke, M. Yukiko, O. Yuko, M. Kennedy and S. Masayuki. 2007, Inverse regulation of leptin mRNA expression by short- and long-chain fatty acids in cultured bovine adipocytes, Domest. Anim.Endocrinol 33:400-407
  34. Arita, Y., S. Kihara, N. Ouchi, M. Takahashi, K. Maeda and K. Miyagawa. 1999, Paradoxical decrease of an adipose-specific protein, adiponectin, in obesity, Biochem. Biophys. Res. Commun 257:79-83
  35. Bensaid, M., M. Gary-Bobo, A. Esclangon, J. P. Maffrand, G. Le- Fur and F. Oury-Donat. 2003, The cannabinoid CB1 receptor antagonist SR141716 increases Acrp30 mRNA expression in adipose tissue of obese fa/fa rats and in cultured adipocyte cells, Mol. Pharmacol 63:908-914
  36. Jacobi, S. K., K. M. Ajuwon, T. E. Weber, J. L. Kuske, C. J. Dyer and M. E. Spurlock. 2004, Cloning and expression of porcine adiponectin, and its relationship to adiposity, lipogenesis and the acute phase response, J. Endocrinol 182:133-144
  37. Motoshima, H., X. Wu, M. K. Sinha, V. E. Hardy, E. L. Rosato and D. J. Barbot. 2002, Differential regulation of adiponectin secretion from cultured human omental and subcutaneous adipocytes: effects of insulin and rosiglitazone, J. Clin. Endocrinol. Metab 87:5662-5667

Cited by

  1. Changes in circulating adiponectin and metabolic hormone concentrations during periparturient and lactation periods in Holstein dairy cows vol.83, pp.12, 2012,
  2. Abundance of adiponectin system and G-protein coupled receptor GPR109A mRNA in adipose tissue and liver of F2 offspring cows of Charolais × German Holstein crosses that differ in body fat accumulation vol.96, pp.1, 2013,