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 (original) (raw)

Abstract

Induction of protein kinase C (PKC) pathway in the vascular tissues by hyperglycemia has been associated with many of the cellular changes observed in the complications of diabetes. Recently, we have reported that the use of a novel, orally effective specific inhibitor of PKC beta isoform (LY333531) normalized many of the early retinal and renal hemodynamics in rat models of diabetes. In the present study, we have characterized a spectrum of biochemical and molecular abnormalities associated with chronic changes induced by glucose or diabetes in the cultured mesangial cells and renal glomeruli that can be prevented by LY333531. Hyperglycemia increased diacylglycerol (DAG) level in cultured mesangial cells exposed to high concentrations of glucose and activated PKC alpha and beta1 isoforms in the renal glomeruli of diabetic rats. The addition of PKC beta selective inhibitor (LY333531) to cultured mesangial cells inhibited activated PKC activities by high glucose without lowering DAG levels and LY333531 given orally in diabetic rats specifically inhibited the activation of PKC beta1 isoform without decreasing PKC alpha isoform activation. Glucose-induced increases in arachidonic acid release, prostaglandin E2 production, and inhibition of Na+-K+ ATPase activities in the cultured mesangial cells were completely prevented by the addition of LY333531. Oral feeding of LY333531 prevented the increased mRNA expression of TGF-beta1 and extracellular matrix components such as fibronectin and alpha1(IV) collagen in the glomeruli of diabetic rats in parallel with inhibition of glomerular PKC activity. These results suggest that the activation of PKC, predominately the beta isoform by hyperglycemia in the mesangial cells and glomeruli can partly contribute to early renal dysfunctions by alteration of prostaglandin production and Na+-K+ ATPase activity as well as the chronic pathological changes by the overexpression of TGF-beta1 and extracellular matrix components genes.

Full Text

The Full Text of this article is available as a PDF (410.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aiello L. P., Robinson G. S., Lin Y. W., Nishio Y., King G. L. Identification of multiple genes in bovine retinal pericytes altered by exposure to elevated levels of glucose by using mRNA differential display. Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):6231–6235. doi: 10.1073/pnas.91.13.6231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ayo S. H., Radnik R. A., Glass W. F., 2nd, Garoni J. A., Rampt E. R., Appling D. R., Kreisberg J. I. Increased extracellular matrix synthesis and mRNA in mesangial cells grown in high-glucose medium. Am J Physiol. 1991 Feb;260(2 Pt 2):F185–F191. doi: 10.1152/ajprenal.1991.260.2.F185. [DOI] [PubMed] [Google Scholar]
  3. Ayo S. H., Radnik R., Garoni J. A., Troyer D. A., Kreisberg J. I. High glucose increases diacylglycerol mass and activates protein kinase C in mesangial cell cultures. Am J Physiol. 1991 Oct;261(4 Pt 2):F571–F577. doi: 10.1152/ajprenal.1991.261.4.F571. [DOI] [PubMed] [Google Scholar]
  4. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  5. Bell R. M., Burns D. J. Lipid activation of protein kinase C. J Biol Chem. 1991 Mar 15;266(8):4661–4664. [PubMed] [Google Scholar]
  6. Bollineni J. S., Reddi A. S. Transforming growth factor-beta 1 enhances glomerular collagen synthesis in diabetic rats. Diabetes. 1993 Nov;42(11):1673–1677. doi: 10.2337/diab.42.11.1673. [DOI] [PubMed] [Google Scholar]
  7. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  8. Brownlee M., Vlassara H., Cerami A. Nonenzymatic glycosylation and the pathogenesis of diabetic complications. Ann Intern Med. 1984 Oct;101(4):527–537. doi: 10.7326/0003-4819-101-4-527. [DOI] [PubMed] [Google Scholar]
  9. Burns D. J., Bloomenthal J., Lee M. H., Bell R. M. Expression of the alpha, beta II, and gamma protein kinase C isozymes in the baculovirus-insect cell expression system. Purification and characterization of the individual isoforms. J Biol Chem. 1990 Jul 15;265(20):12044–12051. [PubMed] [Google Scholar]
  10. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  11. Craven P. A., Caines M. A., DeRubertis F. R. Sequential alterations in glomerular prostaglandin and thromboxane synthesis in diabetic rats: relationship to the hyperfiltration of early diabetes. Metabolism. 1987 Jan;36(1):95–103. doi: 10.1016/0026-0495(87)90070-9. [DOI] [PubMed] [Google Scholar]
  12. Craven P. A., Davidson C. M., DeRubertis F. R. Increase in diacylglycerol mass in isolated glomeruli by glucose from de novo synthesis of glycerolipids. Diabetes. 1990 Jun;39(6):667–674. doi: 10.2337/diab.39.6.667. [DOI] [PubMed] [Google Scholar]
  13. Craven P. A., DeRubertis F. R. Protein kinase C is activated in glomeruli from streptozotocin diabetic rats. Possible mediation by glucose. J Clin Invest. 1989 May;83(5):1667–1675. doi: 10.1172/JCI114066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Craven P. A., Patterson M. C., DeRubertis F. R. Role of enhanced arachidonate availability through phospholipase A2 pathway in mediation of increased prostaglandin synthesis by glomeruli from diabetic rats. Diabetes. 1988 Apr;37(4):429–435. doi: 10.2337/diab.37.4.429. [DOI] [PubMed] [Google Scholar]
  15. Danne T., Spiro M. J., Spiro R. G. Effect of high glucose on type IV collagen production by cultured glomerular epithelial, endothelial, and mesangial cells. Diabetes. 1993 Jan;42(1):170–177. doi: 10.2337/diab.42.1.170. [DOI] [PubMed] [Google Scholar]
  16. Derubertis F. R., 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 Jan;43(1):1–8. doi: 10.2337/diab.43.1.1. [DOI] [PubMed] [Google Scholar]
  17. Ewart H. S., Klip A. Hormonal regulation of the Na(+)-K(+)-ATPase: mechanisms underlying rapid and sustained changes in pump activity. Am J Physiol. 1995 Aug;269(2 Pt 1):C295–C311. doi: 10.1152/ajpcell.1995.269.2.C295. [DOI] [PubMed] [Google Scholar]
  18. Greene D. A., Lattimer S. A., Sima A. A. Sorbitol, phosphoinositides, and sodium-potassium-ATPase in the pathogenesis of diabetic complications. N Engl J Med. 1987 Mar 5;316(10):599–606. doi: 10.1056/NEJM198703053161007. [DOI] [PubMed] [Google Scholar]
  19. Heasley L. E., Johnson G. L. Regulation of protein kinase C by nerve growth factor, epidermal growth factor, and phorbol esters in PC12 pheochromocytoma cells. J Biol Chem. 1989 May 25;264(15):8646–8652. [PubMed] [Google Scholar]
  20. Inoguchi T., Battan R., Handler E., Sportsman J. R., Heath W., King G. L. Preferential elevation of protein kinase C isoform beta II and diacylglycerol levels in the aorta and heart of diabetic rats: differential reversibility to glycemic control by islet cell transplantation. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):11059–11063. doi: 10.1073/pnas.89.22.11059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ishii H., Jirousek M. R., Koya D., Takagi C., Xia P., Clermont A., Bursell S. E., Kern T. S., Ballas L. M., Heath W. F. Amelioration of vascular dysfunctions in diabetic rats by an oral PKC beta inhibitor. Science. 1996 May 3;272(5262):728–731. doi: 10.1126/science.272.5262.728. [DOI] [PubMed] [Google Scholar]
  22. Jirousek M. R., Gillig J. R., Gonzalez C. M., Heath W. F., McDonald J. H., 3rd, Neel D. A., Rito C. J., Singh U., Stramm L. E., Melikian-Badalian A. (S)-13-[(dimethylamino)methyl]-10,11,14,15-tetrahydro-4,9:16, 21-dimetheno-1H, 13H-dibenzo[e,k]pyrrolo[3,4-h][1,4,13]oxadiazacyclohexadecene-1,3(2H)-d ione (LY333531) and related analogues: isozyme selective inhibitors of protein kinase C beta. J Med Chem. 1996 Jul 5;39(14):2664–2671. doi: 10.1021/jm950588y. [DOI] [PubMed] [Google Scholar]
  23. Kikkawa R., Haneda M., Uzu T., Koya D., Sugimoto T., Shigeta Y. Translocation of protein kinase C alpha and zeta in rat glomerular mesangial cells cultured under high glucose conditions. Diabetologia. 1994 Aug;37(8):838–841. doi: 10.1007/BF00404342. [DOI] [PubMed] [Google Scholar]
  24. Kikkawa R., Umemura K., Haneda M., Arimura T., Ebata K., Shigeta Y. Evidence for existence of polyol pathway in cultured rat mesangial cells. Diabetes. 1987 Feb;36(2):240–243. doi: 10.2337/diab.36.2.240. [DOI] [PubMed] [Google Scholar]
  25. Koya D., Kikkawa R., Haneda M., Uzu T., Sawada M., Kajiwara N., Sakamoto K., Sugimoto T., Shigeta Y. Nipradilol inhibits rat mesangial cell mitogenesis through the activation of soluble guanylate cyclase. Eur J Pharmacol. 1993 Mar 15;245(1):79–82. doi: 10.1016/0922-4106(93)90173-7. [DOI] [PubMed] [Google Scholar]
  26. Kraft A. S., Anderson W. B. Phorbol esters increase the amount of Ca2+, phospholipid-dependent protein kinase associated with plasma membrane. Nature. 1983 Feb 17;301(5901):621–623. doi: 10.1038/301621a0. [DOI] [PubMed] [Google Scholar]
  27. Kurkinen M., Condon M. R., Blumberg B., Barlow D. P., Quinones S., Saus J., Pihlajaniemi T. Extensive homology between the carboxyl-terminal peptides of mouse alpha 1(IV) and alpha 2(IV) collagen. J Biol Chem. 1987 Jun 25;262(18):8496–8499. [PubMed] [Google Scholar]
  28. Mauer S. M., Steffes M. W., Ellis E. N., Sutherland D. E., Brown D. M., Goetz F. C. Structural-functional relationships in diabetic nephropathy. J Clin Invest. 1984 Oct;74(4):1143–1155. doi: 10.1172/JCI111523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mitchell F. E., Marais R. M., Parker P. J. The phosphorylation of protein kinase C as a potential measure of activation. Biochem J. 1989 Jul 1;261(1):131–136. doi: 10.1042/bj2610131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Negrete H., Studer R. K., Craven P. A., DeRubertis F. R. Role for transforming growth factor beta in thromboxane-induced increases in mesangial cell fibronectin synthesis. Diabetes. 1995 Mar;44(3):335–339. doi: 10.2337/diab.44.3.335. [DOI] [PubMed] [Google Scholar]
  31. Newton A. C. Protein kinase C: structure, function, and regulation. J Biol Chem. 1995 Dec 1;270(48):28495–28498. doi: 10.1074/jbc.270.48.28495. [DOI] [PubMed] [Google Scholar]
  32. Nishizuka Y. The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature. 1988 Aug 25;334(6184):661–665. doi: 10.1038/334661a0. [DOI] [PubMed] [Google Scholar]
  33. Ohno S., Konno Y., Akita Y., Yano A., Suzuki K. A point mutation at the putative ATP-binding site of protein kinase C alpha abolishes the kinase activity and renders it down-regulation-insensitive. A molecular link between autophosphorylation and down-regulation. J Biol Chem. 1990 Apr 15;265(11):6296–6300. [PubMed] [Google Scholar]
  34. Preiss J. E., Loomis C. R., Bell R. M., Niedel J. E. Quantitative measurement of sn-1,2-diacylglycerols. Methods Enzymol. 1987;141:294–300. doi: 10.1016/0076-6879(87)41077-x. [DOI] [PubMed] [Google Scholar]
  35. Pugliese G., Pricci F., Pugliese F., Mene P., Lenti L., Andreani D., Galli G., Casini A., Bianchi S., Rotella C. M. Mechanisms of glucose-enhanced extracellular matrix accumulation in rat glomerular mesangial cells. Diabetes. 1994 Mar;43(3):478–490. doi: 10.2337/diab.43.3.478. [DOI] [PubMed] [Google Scholar]
  36. Schambelan M., Blake S., Sraer J., Bens M., Nivez M. P., Wahbe F. Increased prostaglandin production by glomeruli isolated from rats with streptozotocin-induced diabetes mellitus. J Clin Invest. 1985 Feb;75(2):404–412. doi: 10.1172/JCI111714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shankland S. J., Scholey J. W., Ly H., Thai K. Expression of transforming growth factor-beta 1 during diabetic renal hypertrophy. Kidney Int. 1994 Aug;46(2):430–442. doi: 10.1038/ki.1994.291. [DOI] [PubMed] [Google Scholar]
  38. Sharma K., Jin Y., Guo J., Ziyadeh F. N. Neutralization of TGF-beta by anti-TGF-beta antibody attenuates kidney hypertrophy and the enhanced extracellular matrix gene expression in STZ-induced diabetic mice. Diabetes. 1996 Apr;45(4):522–530. doi: 10.2337/diab.45.4.522. [DOI] [PubMed] [Google Scholar]
  39. Sharma K., Ziyadeh F. N. Hyperglycemia and diabetic kidney disease. The case for transforming growth factor-beta as a key mediator. Diabetes. 1995 Oct;44(10):1139–1146. doi: 10.2337/diab.44.10.1139. [DOI] [PubMed] [Google Scholar]
  40. Sharma K., Ziyadeh F. N. Renal hypertrophy is associated with upregulation of TGF-beta 1 gene expression in diabetic BB rat and NOD mouse. Am J Physiol. 1994 Dec;267(6 Pt 2):F1094–F1001. doi: 10.1152/ajprenal.1994.267.6.F1094. [DOI] [PubMed] [Google Scholar]
  41. Shiba T., Inoguchi T., Sportsman J. R., Heath W. F., Bursell S., King G. L. Correlation of diacylglycerol level and protein kinase C activity in rat retina to retinal circulation. Am J Physiol. 1993 Nov;265(5 Pt 1):E783–E793. doi: 10.1152/ajpendo.1993.265.5.E783. [DOI] [PubMed] [Google Scholar]
  42. Steffes M. W., Bilous R. W., Sutherland D. E., Mauer S. M. Cell and matrix components of the glomerular mesangium in type I diabetes. Diabetes. 1992 Jun;41(6):679–684. doi: 10.2337/diab.41.6.679. [DOI] [PubMed] [Google Scholar]
  43. Studer R. K., Craven P. A., DeRubertis F. R. Role for protein kinase C in the mediation of increased fibronectin accumulation by mesangial cells grown in high-glucose medium. Diabetes. 1993 Jan;42(1):118–126. doi: 10.2337/diab.42.1.118. [DOI] [PubMed] [Google Scholar]
  44. Vasilets L. A., Schwarz W. Structure-function relationships of cation binding in the Na+/K(+)-ATPase. Biochim Biophys Acta. 1993 Oct 29;1154(2):201–222. doi: 10.1016/0304-4157(93)90012-d. [DOI] [PubMed] [Google Scholar]
  45. Wakisaka M., Spiro M. J., Spiro R. G. Synthesis of type VI collagen by cultured glomerular cells and comparison of its regulation by glucose and other factors with that of type IV collagen. Diabetes. 1994 Jan;43(1):95–103. doi: 10.2337/diab.43.1.95. [DOI] [PubMed] [Google Scholar]
  46. Williams B., Schrier R. W. Characterization of glucose-induced in situ protein kinase C activity in cultured vascular smooth muscle cells. Diabetes. 1992 Nov;41(11):1464–1472. doi: 10.2337/diab.41.11.1464. [DOI] [PubMed] [Google Scholar]
  47. Williams B., Schrier R. W. Glucose-induced protein kinase C activity regulates arachidonic acid release and eicosanoid production by cultured glomerular mesangial cells. J Clin Invest. 1993 Dec;92(6):2889–2896. doi: 10.1172/JCI116911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Williamson J. R., Chang K., Frangos M., Hasan K. S., Ido Y., Kawamura T., Nyengaard J. R., van den Enden M., Kilo C., Tilton R. G. Hyperglycemic pseudohypoxia and diabetic complications. Diabetes. 1993 Jun;42(6):801–813. doi: 10.2337/diab.42.6.801. [DOI] [PubMed] [Google Scholar]
  49. Xia P., Kramer R. M., King G. L. Identification of the mechanism for the inhibition of Na+,K(+)-adenosine triphosphatase by hyperglycemia involving activation of protein kinase C and cytosolic phospholipase A2. J Clin Invest. 1995 Aug;96(2):733–740. doi: 10.1172/JCI118117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Yamamoto T., Nakamura T., Noble N. A., Ruoslahti E., Border W. A. Expression of transforming growth factor beta is elevated in human and experimental diabetic nephropathy. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1814–1818. doi: 10.1073/pnas.90.5.1814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Yasuda I., Kishimoto A., Tanaka S., Tominaga M., Sakurai A., Nishizuka Y. A synthetic peptide substrate for selective assay of protein kinase C. Biochem Biophys Res Commun. 1990 Feb 14;166(3):1220–1227. doi: 10.1016/0006-291x(90)90996-z. [DOI] [PubMed] [Google Scholar]
  52. Zhang J., Wang L., Petrin J., Bishop W. R., Bond R. W. Characterization of site-specific mutants altered at protein kinase C beta 1 isozyme autophosphorylation sites. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6130–6134. doi: 10.1073/pnas.90.13.6130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Ziyadeh F. N., Sharma K., Ericksen M., Wolf G. Stimulation of collagen gene expression and protein synthesis in murine mesangial cells by high glucose is mediated by autocrine activation of transforming growth factor-beta. J Clin Invest. 1994 Feb;93(2):536–542. doi: 10.1172/JCI117004. [DOI] [PMC free article] [PubMed] [Google Scholar]