Positive and Negative Feedback Regulation in the Production and Secretion of Insulin from INS-1 Cells by Testosterone

Abstract

Type 2 diabetes is often developed in genetically predisposed subjects combined with sedentary life style or environmental factors. In women with polycystic ovary syndrome, hyperandrogenism is often accompanied with hyperinsulinemia and insulin resistance. Further, some studies have found associations of hyperandrogenemia with β-cell dysfunction and type 2 diabetes. We therefore tested the impairment effect of testosterone on glucose-stimulated insulin secretion in INS-1 cells. INS-1 cells were treated with different concentrations of testosterone and examined at different time points. In contrast to control, excess testosterone treatment for 48 h could promote glucose-stimulated insulin secretion and enhance pancreatic/duodenal homeobox-1 and glucose transporter-2 mRNA expression up to 2-fold. Alternatively, long-time and high-concentration testosterone treatment significantly impaired glucose-stimulated insulin secretion and insulin mRNA levels and promoted malondialdehyde content. Androgen receptor antagonist flutamide could partly attenuate glucose-stimulated insulin secretion. These results indicate that direct in vitro exposure of INS-1 cells to testosterone had both concentration- and time-dependent effects on glucose-stimulated insulin secretion, gene expression, and oxidative stress. These findings showed to some extent that excess circulation of testosterone might impair β-cell function, and further contribute to the etiology of insulin resistance in polycystic ovary syndrome.

Key words

testosterone - INS-1 cells - GSIS

*

^* These authors contributed equally to this work.

• References

• 1 Abbott DH, Bruns CR, Barnett DK, Dunaif A, Goodfriend TL, Dumesic DA, Tarantal AF. Experimental induced gestational androgen excess disrupts glucoregulation in rhesus monkey dams and their female offspring. Am J Physiol Endocrinol Metab 2010; 299: E741-E751
• 2 Lazic M, Aird F, Levine JE, Dunaif A. Prenatal androgen treatment alters body composition and glucose homeostasis in male rats. J Endocrinol 2011; 208: 293-300
• 3 Grillo ML, Jacobus AP, Scalco R, Amaral F, Rodrigues DO, Loss ES, Wassermann GF. Testosterone rapidly stimulates insulin releases from isolated pancreatic islets through a non-genomic dependent mechanism. Horm Meta Res 2005; 37: 662-665
• 4 Porte Jr. D. Banting lecture 1990. Beta-cells in type II diabetes mellitus. Diabetes 1991; 40: 166-180
• 5 Kaneto H, Miyatsuka T, Shiraiwa T, Yamamoto K, Kato K, Fujitani Y, Matsuoka TA. Crucial role of PDX-1 in pancreas development, beta-cell differentiation, and induction of surrogate beta-cells. Curr Med Chem 2007; 14: 1745-1752
• 6 Jonsson J, Carlsson L, Edlund T, Edlund H. Insulin-promoter-factor 1 is required for pancreas development in mice. Nature 1994; 371: 606-609
• 7 Dutta S, Bonner-Weir S, Montminy M, Wright C. Regulatory factor linked to late-onset diabetes?. Nature 1998; 392: 560
• 8 Macfarlane WM, Read ML, Gilligan M, Bujalska I, Docherty K. Glucose modulates the binding activity of the beta-cell transcription factor IUF1 in a phosphorylation-dependent manner. Biochem J 1994; 303: 625-631
• 9 Marshak S, Totary H, Cerasi E, Melloul D. Purification of the beta-cell glucose-sensitive factor that transactivates the insulin gene differentially in normal and transformed islet cells. Proc Natl Acad Sci USA 1996; 93: 15057-15062
• 10 Macfarlane WM, McKinnon CM, Felton-Edkins ZA, Cragg H, James RF, Docherty K. Glucose stimulates translocation of the homeodomain transcription factor PDX1 from the cytoplasm to the nucleus in pancreatic beta-cells. J Biol Chem 1999; 274: 1011-1016
• 11 Petersen HV, Peshavaria M, Pedersen AA, Philippe J, Stein R, Madsen OD, Serup P. Glucose stimulates the activation domain potential of the PDX-1 homeodomain transcription factor. FEBS Lett 1998; 431: 362-366
• 12 Tiedge M, Lenzen S. Regulation of glucokinase and GLUT-2 glucose-transporter gene expression in pancreatic β cells. Biochem J 1991; 279: 899-901
• 13 Lawrence MC, Jivan A, Shao C, Duan L, Goad D, Zaganjor E, Osborne J, McGlynn K, Stippec S, Earnest S, Chen W, Cobb MH. The roles of MAPKs in disease. Cell Res 2008; 18: 436-442
• 14 Khoo S, Griffen SC, Xia Y, Baer RJ, German MS, Cobb MH. Regulation of insulin gene transcription by ERK1 and ERK2 in pancreatic beta cells. J Biol Chem 2003; 278: 32969-32977
• 15 Lawrence MC, McGlynn K, Park BH, Cobb MH. ERK1/2-dependent activation of transcription factors required for acute and chronic effects of glucose on the insulin gene promoter. J Biol Chem 2005; 280: 26751-26759
• 16 Benes C, Poitout V, Marie JC, Martin-Perez J, Roisin MP, Fagard R. Mode of regulation of the extracellular signal-regulated kinases in the pancreatic beta-cell line MIN6 and their implication in the regulation of insulin gene transcription. Biochem J 1999; 340: 219-225
• 17 Diaz-Sanchez V, Morimoto S, Morales A, Robles-Diaz G, Cerbon M. Androgen receptor in the rat pancreas: genetic expression and steroid regulation. Pancreas 1995; 11: 241-245
• 18 Cato AC, Peterziel H. The androgen receptor as mediator of gene expression and signal transduction pathways. Trends Endocrinol Metab 1998; 9: 150-154
• 19 Melloul D, Ben-Neriah Y, Cerasi E. Glucose modulates the binding of an islet-specific factor to a conserved sequence within the rat I and the human insulin promoters. Proc Natl Acad Sci USA 1993; 90: 3865-3869
• 20 Peers B, Leonard J, Sharma S, Teitelman G, Montminy MR. Insulin expression in pancreatic islet cells relies on cooperative interactions between the helix loop helix factor E47 and the homeobox factor STF-1. Mol Endocrinol 1994; 8: 1798-1806
• 21 Robertson RP, Harmon J, Tran PO, Tanaks Y, Takahashi H. Glucose toxicity in beta-cells: type 2 diabetes, good radicals gone bad, and the glutathione connection. Diabetes 2003; 52: 581-587
• 22 Morimoto S, Fernandez-Mejia C, Romero-Navarro G, Morales-Peza N, Díaz-Sánchez V. Testosterone effect on insulin content, messenger ribonucleic acid levels, promoter activity, and secretion in the rat. Endocrinology 2001; 142: 1442-1447
• 23 Medina MC, Souza LC, Caperuto LC, Anhe GF, Amanso AM, Teixeira VP, Bordin S, Carpinelli AR, Britto LR, Barbieri RL, Borella MI, Carvalho CR. Dehydroepiandrosterone increases beta-cell mass and improves the glucose-induced insulin secretion by pancreatic islets from aged rats. FEBS Lett 2006; 580: 285-290
• 24 Del Guerra S, Lupi R, Marselli L, Masini M, Bugliani M, Sbrana S, Torri S, Pollera M, Boggi U, Mosca F, Del Prato S, Marchetti P. Functional and molecular defects of pancreatic islets in human type 2 diabetes. Diabetes 2005; 54: 727-735
• 25 Ripple MO, Hagopian K, Oberley TD, Schatten H, Weindruch R. Androgen-induced oxidative stress in human LNCaP prostate cancer cells is associated with multiple mitochondrial modifications. Antioxid Redox Signal 1999; 1: 71-81
• 26 Pansarasa O, D’Antona G, Gualea MR, Marzani B, Pellegrino MA, Marzatico F. “Oxidative stress”: effects of mild endurance training and testosterone treatment on rat gastrocnemius muscle. Eur J Appl Physiol 2002; 87: 550-555
• 27 Gonzalez F, Rote NS, Minium J, Kirwan JP. Reactive oxygen species-induced oxidative stress in the development of insulin resistance and hyperandrogenism in polycystic ovary syndrome. J Clin Endocrinol Metab 2006; 91: 336-340
• 28 Kan WH, Hsieh CH, Schwacha MG, Choudhry MA, Raju R, Bland KI, Chaudry IH. Flutamide protects against trauma-hemorrhage-induced liver injury via attenuation of the inflammatory response, oxidative stress, and apopotosis. J Appl Physiol 2008; 105: 595-602
• 29 Pi J, Collins S. Reactive oxygen species and uncoupling protein 2 in pancreatic beta-cell function. Diabetes Obes Metab 2010; 12 (Suppl. 02) 141-148
• 30 Kawamori D, Kajimoto Y, Kaneto H, Umayahara Y, Fujitani Y, Miyatsuka T, Watada H, Leibiger IB, Yamasaki Y, Hori M. Oxidative stress induces nucleo-cytoplasmic translocation of pancreatic transcription factor PDX-1 through activation of c-Jun NH(2)-terminal kinase. Diabetes 2003; 52: 2896-2904
• 31 Chen Z, Gibson TB, Robinson F, Silvestro L, Pearson G, Xu B, Wright A, Vanderbilt C, Cobb MH. MAP kinases. Chem Rev 2001; 101: 2449-2476
• 32 Arnette D, Gibson TB, Lawrence MC, January B, Khoo S, McGlynn K, Vanderbilt CA, Cobb MH. Regulation of ERK1 and ERK2 by glucose and peptide hormones in pancreatic beta cells. J Biol Chem 2003; 278: 32517-32525
• 33 Lawrence MC, McGlynn K, Park BH, Cobb MH. ERK1/2-dependent activation of transcription factors required for acute and chronic effects of glucose on the insulin gene promoter. J Biol Chem 2005; 280: 26751-26759
• 34 Marshak S, Benshushan E, Shoshkes M, Havin L, Cerasi E, Melloul D. Functional conservation of regulatory elements in the pdx-1 gene: PDX-1 and hepatocyte nuclear factor 3 beta transcription factors mediate beta-cell-specific expression. Mol Cell Biol 2000; 20: 7583-7590
• 35 Zangen DH, Bonner-Weir S, Lee CH, Latimer JB, Miller CP, Habener JF, Weir GC. Reduced insulin, GLUT2, and IDX-1 in beta-cells after partial pancreatectomy. Diabetes 1997; 46: 258-264
• 36 Pelekanou V, Notas G, Sanidas E, Tsapis A, Castanas E, Kampa M. Testosterone membrane – initiated action in breast cancer cells: Interaction with the androgen signaling pathway and EPOR. Mol Oncol 2010; 4: 135-149