The relationship between high glucose-induced secretion of IGFs and PKC or oxidative stress in mesangial cells

Mesangial 세포에서 고포도당에 의한 IGFs 분비와 PKC 및 산화성 스트레스와의 관련성에 관한 연구

  • Park, Su-hyun (Department of Veterinary Physiology, College of Veterinary Medicine, Chonnam National University) ;
  • Heo, Jung-sun (Department of Veterinary Physiology, College of Veterinary Medicine, Chonnam National University) ;
  • Kang, Chang-won (Bio-safety Research Institute, College of Veterinary Medicine, Chonbuk National University) ;
  • Han, Ho-jae (Department of Veterinary Physiology, College of Veterinary Medicine, Chonnam National University)
  • 박수현 (전남대학교 수의과대학 생리학교실) ;
  • 허정선 (전남대학교 수의과대학 생리학교실) ;
  • 강창원 (전북대학교 수의과대학 생리학교실, 생체안전성 연구소) ;
  • 한호재 (전남대학교 수의과대학 생리학교실)
  • Accepted : 2004.11.08
  • Published : 2004.12.30


The proliferation of mesangial cells has been associated with the development of diabetic nephropathy. The cell proliferation has been regulated by diverse growth factors. Among them, insulin like growth factors(IGFs) are also involved in the pathogenesis of diabetic nephropathy. However, it is not yet known about the effect of high glucose on IGF-I and IGF-II secretion and the relationship between high glucose-induced secretion of IGFs and PKC or oxidative stress in the mesangial cells. Thus, we examined the mechanisms by which high glucose regulates secretion of IGFs in mesangial cells. High glucose(25 mM) increased IGF-I and IGF-II secretion. High glucose-induced increase of IGF-I and IGF-II secretion were blocked by taurine($2{\times}10^{-3}$ M), N-acetyl cystein(NAC, $10^{-5}M$), or GSH($10^{-5}M$) (antioxidants), suggesting the role of oxidative stress. High glucose-induced secretion of IGF-I and IGF-II were blocked by H-7, staurosporine, and bisindolylmaleimide I(protein kinase C inhibitors). On the other hand, high glucose also increased lipid peroxide (LPO) formation in a dose dependent manner. In addition, high glucoseinduced stimulation of LPO formation was blocked by PKC inhibitors. These results suggest that PKC is responsible for the increase of oxidative stress in the action of high glucose-induced secretion of IGF-I and IGF-II in mesangial cells. In conclusion, high glucose stimulates IGF-I and IGF-II secretion via PKCoxidative stress signal pathways in mesangial cells.


Supported by : 한국학술진흥재단


  1. Abou-Seif, M. A. and Youssef, A. A. Oxidative stress and male IGF-1, gonadotropin and related hormones in diabetic patients. Clin. Chem. Lab. Med. 2001, 39, 618-623
  2. Agardh, C. D., Stenram, U., Torffvit, O. and Agardh, E. R. Effects of inhibition of glycation and oxidative stress on the development of diabetic nephropathy in rats. J. Diabetes Complications. 2002, 16, 395-400
  3. Babazono, T., Kapor-Drezgic, J., Dlugosz, J. A. and Whiteside, C. Altered expression and subcellular localization of diacylglycerol-sensitive protein kinase C isoforms in diabetic rat glomerular cells. Diabetes. 1998, 47, 668-676
  4. Berfield, A. K., Spicer, D. and Abrass, C. K. Insulinlike growth factor I (IGF-I) induces unique effects in the cytoskeleton of cultured rat glomerular mesangial cells. J. Histochem. Cytochem. 1997, 45, 583-593
  5. Binoux, M. The IGF system in metabolism regulation. Diabetes Metab. 1995, 21, 330-337
  6. Bowsher, R. R., Lee, W. H., Apathy, J. M., O'Brien,P. J., Ferguson, A. L. and Henry, D. P. Measurement of insulin-like growth factor-II in physiological fluids and tissues. I. An improved extraction procedure and radioimmunoassay for human and rat fluids. Endocrinology. 1991, 128, 805-814
  7. Catherwood, M. A., Powell, L. A., Anderson, P., McMaster, D., Sharpe, P. C. and Trimble, E. R. Glucose-induced oxidative stress in mesangial cells. Kidney Int. 2002, 61, 599-608
  8. Craven, P. A., Davidson, C. M. and DeRubertis, F. R. Increase in diacylglycerol mass in isolated glomeruli by glucose from de novo synthesis of glycerolipids. Diabetes. 1990, 39, 667-674
  9. Daughaday, W. H. and Rotwein, P. Insulin-like growth factors I and II. Peptide messenger ribonucleic acid and gene structure serum, and tissue concentrations. Endocr. Rev. 1989, 10, 68-91
  10. De La Puente, A., Goya, L., Ramos, S., Martin, M. A., Alvarez, C., Escriva, F. and Pascual-Leone, A. M. Effects of experimental diabetes on renal IGF/IGFBP system during neonatal period in the rat. Am. J. Physiol. Renal. Physiol. 2000, 279, F1067-F1076
  11. Derubertis, F. R. and Craven, P. A. Activation of protein kinase C in glomerular cells in diabetes. Mechanisms and potential links to the pathogenesis of diabetic glomerulopathy. Diabetes. 1994, 43, 1-8
  12. Elliot, S. J., Striker, L. J., Hattori, M., Yang, C. W., He, C. J., Peten, E. P. and Striker, G. E. Mesangial cells from diabetic NOD mice constitutively secrete increased amounts of insulin-like growth factor-I. Endocrinology. 1993, 133, 1783-1788
  13. Feld, S. and Hirschberg, R. Growth hormone, the insulin-like growth factor system, and the kidney. Endocr Rev. 1996, 17, 423-480
  14. Flyvbjerg, A., Landau, D., Domene, H., Hernandez, L., Gronbaek, H. and LeRoith, D. The role of growth hormone, insulin-like growth factors (IGFs), and IGFbinding proteins in experimental diabetic kidney disease. Metabolism. 1995, 44, 67-71
  15. Gooch, J. L., Tang, Y., Ricono, J. M. and Abboud, H. E. Insulin-like growth factor-I induces renal cell hypertrophy via a calcineurin-dependent mechanism. J. Biol. Chem. 2001, 276, 42492-42500
  16. Gronbaek, H., Nielsen, B., Frystyk, J., Flyvbjerg, A. and Orskov, H. Effect of lanreotide on local kidney IGF-I and renal growth in experimental diabetes in the rat. Exp. Nephrol. 1996, 4, 295-303
  17. Ha, H., Yu, M. R., Choi, Y. J, Kitamura, M. and Lee, H. B. Role of high glucose-induced nuclear factorkappaB activation in monocyte chemoattractant protein-1 expression by mesangial cells. J. Am. Soc. Nephrol. 2002, 13, 894-902
  18. Heo, Y. R., Kang, C. W. and Cha, Y. S. L-Carnitine changes the levels of insulin-like growth factors (IGFs) and IGF binding proteins in streptozotocin-induced diabetic rat. J. Nutr. Sci. Vitaminol. 2001, 47, 329-334
  19. Hoog, A., Sandberg-Nordqvist, A. C., Abdel-Halim, S. M,, Carlsson-Skwirut, C., Guenifi, A., Tally, M., Ostenson, C. G., Falkmer, S., Sara, V. R., Efendic, S., Schalling, M. and Grimelius, L. Increased amounts of a high molecular weight insulin-like growth factor II (IGF-II) peptide and IGF-II messenger ribonucleic acid in pancreatic islets of diabetic Goto-Kakizaki rats. Endocrinology. 1996, 137, 2415-2423
  20. Inoguchi, T., Li, P., Umeda, F., Yu, H. Y., Kakimoto, M., Imamura, M., Aoki, T., Etoh, T., Hashimoto, T., Naruse, M., Sano, H., Utsumi, H. and Nawata, H. High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase Cdependent activation of NAD(P)H oxidase in cultured vascular cells. Diabetes. 2000, 49, 1939-1945
  21. Iori, E., Marescotti, M. C., Vedovato, M., Ceolotto, G., Avogaro, A., Tiengo, A., Del Prato, S. and Trevisan, R. In situ protein Kinase C activity is increased in cultured fibroblasts from Type 1 diabetic patients with nephropathy. Diabetologia. 2003, 46, 524-530
  22. Kimura, M., Ishizawa, M., Miura, A., Itaya, S., Kanoh, Y., Yasuda, K., Uno, Y., Morita, H. and Ishizuka, T. Platelet protein kinase C isoform content in type 2 diabetes complicated with retinopathy and nephropathy. Platelets. 2001, 12, 138-143
  23. Koya, D., Jirousek, M. R., Lin, Y. W., Ishii, H., Kuboki, K. and King, G. L. Characterization of protein kinase C beta isoform activation on the gene expression of transforming growth factor-beta, extracellular matrix components, and prostanoids in the glomeruli of diabetic rats. J. Clin. Invest. 1997, 100, 115-126
  24. Kumar, A., Hawkins, K. S., Hannan, M. A. and Ganz, M. B. Activation of PKC-beta(I) in glomerular mesangial cells is associated with specific NF-kappaB subunit translocation. Am. J. Physiol. Renal. Physiol. 2001, 281, F613-F619
  25. Kurzawa, R., Glabowski, W., Baczkowski, T. and Brelik, P. Evaluation of mouse preimplantation embryos exposed to oxidative stress cultured with insulin-like growth factor I and II, epidermal growth factor, insulin, transferrin, and selenium. Reprod. Biol. 2002, 2, 143-162
  26. Lee, C. Y. and Henricks, D. M. Comparisions of various acidic treatments of bovine serum on insulinlike growth factor-I immunoreactive and binding activity. J. Endocrinol, 1990, 127, 139-148
  27. Lee, H. B., Yu, M. R., Yang, Y., Jiang, Z. and Ha, H. Reactive oxygen species-regulated signaling pathways in diabetic nephropathy. J. Am. Soc. Nephrol. 2003, 14, S241-S245
  28. Mathews, L. S., Norstedt, G. and Palmiter, R. D. Regulation of insulin-like growth factor I gene expression by growth hormone. Proc. Natl. Acad. Sci. USA. 1986, 83, 9343-9347
  29. Miyatake, N., Shikata, K., Wada, J., Sugimoto, H., Takahashi, S. and Makino, H. Differential distribution of insulin-like growth factor-1 and insulin-like growth factor binding proteins in experimental diabetic rat kidney. Nephron. 1999, 81, 317-323
  30. Nishikawa, T., Edelstein, D., Du, X. L., Yamagishi, S., Matsumura, T., Kaneda, Y., Yorek, M. A. Beebe, D., Oates, P. J., Hammes, H. P., Giardino, I. and Brownlee, M. Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature. 2000, 404, 787-790
  31. Obrosova, I. G., Fathallah, L., Liu, E. and Nourooz- Zadeh, J. Early oxidative stress in the diabetic kidney: effect of DL-alpha-lipoic acid. Free. Radic. Biol. Med. 2003, 34, 186-195
  32. Ohkawa, H., Ohishi, N. and Yagi, K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal. Biochem. 1979, 95, 351-358
  33. Rossert, J., Terraz-Durasnel, C. and Brideau, G. Growth factors, cytokines, and renal fibrosis during the course of diabetic nephropathy. Diabetes. Metab. 2000, 26, S16-S24
  34. Schleicher, E. D. and Olgemoller, B. Glomerular changes in diabetes mellitus. Eur. J. Clin. Chem. Clin. Biochem. 1992, 30, 635-640
  35. Schwander, J. C., Hauri, C., Zapf, J. and Froesch, E. R. Synthesis and secretion of insulin-like growth factor and its binding protein by the perfused rat liver: dependence on growth hormone status. Endocrinology. 1983, 113, 297-305
  36. Sheetz, M. J. and King, G. L. Molecular understanding of hyperglycemia’s adverse effects for diabetic complications. JAMA. 2002, 288, 2579-2588
  37. Spranger, J., Buhnen, J., Jansen, V., Krieg, M., Meyer-Schwickerath, R., Blum, W. F., Schatz, H. and Pfeiffer, A. F. Systemic levels contribute significantly to increased intraocular IGF-I, IGF-II and IGF-BP3 [correction of IFG-BP3] in proliferative diabetic retinopathy. Horm. Metab. Res. 2000, 32, 196-200
  38. Steff, M.W., Osterby, W. R., Chavers, B. and Mauer, S. M. Mesangial expansion as a central mechanism for loss of kidney function in diabetic patients. Diabetes. 1998, 38, 1077-1081
  39. Studer, R. K, Craven, P. A. and DeRubertis, F. R. Antioxidant inhibition of protein kinase C-signaled increases in transforming growth factor-beta in mesangial cells. Metabolism. 1997, 46, 918-925
  40. Tack, I., Elliot, S. J., Potier, M., Rivera, A., Striker, G. E. and Striker, L. J. Autocrine activation of the IGF-I signaling pathway in mesangial cells isolated from diabetic NOD mice. Diabetes. 2002, 51, 182-188
  41. Tuttle, K. R. and Anderson, P. W. A novel potential therapy for diabetic nephropathy and vascular complications: protein kinase C beta inhibition. Am. J. Kidney Dis. 2003, 42, 456-465
  42. Wardle, E. N. How does hyperglycaemia predispose to diabetic nephropathy? QJM. 1996, 89, 943-951
  43. Werner, H., Shen-Orr, Z., Stannard, B., Burguera, B., Roberts, C. T. Jr. and LeRoith, D. Experimental diabetes increases insulin-like growth factor I and II receptor concentration and gene expression in kidney. Diabetes. 1990, 39, 1490-1497
  44. West, I. C. Radicals and oxidative stress in diabetes. Diabet Med. 2000, 17, 171-180
  45. Yakar, S., Wu, Y., Setser, J. and Rosen, C. J. The role of circulating IGF-I: lessons from human and animal models. Endocrine. 2002, 19, 239-248
  46. Zhuang, H. X., Wuarin, L., Fei, Z. J. and Ishii, D. N. Insulin-like growth factor (IGF) gene expression is reduced in neural tissues and liver from rats with noninsulin-dependent diabetes mellitus, and IGF treatment ameliorates diabetic neuropathy. J. Pharmacol. Exp. Ther. 1997, 283, 366-374