Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1993 Jun 1;90(11):4947–4951. doi: 10.1073/pnas.90.11.4947

Elevated glucose and angiotensin II increase 12-lipoxygenase activity and expression in porcine aortic smooth muscle cells.

R Natarajan 1, J L Gu 1, J Rossi 1, N Gonzales 1, L Lanting 1, L Xu 1, J Nadler 1
PMCID: PMC46630  PMID: 8506339

Abstract

The lipoxygenase (LO) pathway of arachidonate metabolism has been suggested to play a key role in atherosclerosis and in mediating several actions of angiotensin II (AII). However, the relationship between LO activation and factors linked to accelerated diabetic vascular disease such as hyperglycemia and AII is not known. We have investigated the effect of high glucose (HG; 25 mM) and AII on LO activity as well as LO protein and mRNA expression in porcine aortic vascular smooth muscle cells (PVSMCs). We observed that cells cultured in HG had significantly higher levels of the cell-associated LO products 12- and 15-hydroxyeicosatetraenoic acids (HETEs). AII added to cells grown in HG specifically further increased only cell-associated 12-HETE levels. Using immunoblot analysis and reverse transcriptase PCRs, we demonstrated the presence in PVSMCs of porcine leukocyte-type 12-LO protein and mRNA, respectively. Furthermore, the levels of both were markedly upregulated by AII as well as by HG. These studies suggest that enhanced 12-LO activity and expression are mechanisms for accelerated vascular disease produced by HG and AII in diabetes mellitus.

Full text

PDF
4947

Images in this article

4950Fig. 4
on p.4950
4950Fig. 5
on p.4950
4950Fig. 6
on p.4950
4951Fig. 7
on p.4951

Selected References

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

  1. Arakawa T., Oshima T., Kishimoto K., Yoshimoto T., Yamamoto S. Molecular structure and function of the porcine arachidonate 12-lipoxygenase gene. J Biol Chem. 1992 Jun 15;267(17):12188–12191. [PubMed] [Google Scholar]
  2. Brown M. L., Jakubowski J. A., Leventis L. L., Deykin D. Elevated glucose alters eicosanoid release from porcine aortic endothelial cells. J Clin Invest. 1988 Dec;82(6):2136–2141. doi: 10.1172/JCI113835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Daemen M. J., Lombardi D. M., Bosman F. T., Schwartz S. M. Angiotensin II induces smooth muscle cell proliferation in the normal and injured rat arterial wall. Circ Res. 1991 Feb;68(2):450–456. doi: 10.1161/01.res.68.2.450. [DOI] [PubMed] [Google Scholar]
  4. De Marzo N., Sloane D. L., Dicharry S., Highland E., Sigal E. Cloning and expression of an airway epithelial 12-lipoxygenase. Am J Physiol. 1992 Feb;262(2 Pt 1):L198–L207. doi: 10.1152/ajplung.1992.262.2.L198. [DOI] [PubMed] [Google Scholar]
  5. Funk C. D., Furci L., FitzGerald G. A. Molecular cloning, primary structure, and expression of the human platelet/erythroleukemia cell 12-lipoxygenase. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5638–5642. doi: 10.1073/pnas.87.15.5638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Geisterfer A. A., Peach M. J., Owens G. K. Angiotensin II induces hypertrophy, not hyperplasia, of cultured rat aortic smooth muscle cells. Circ Res. 1988 Apr;62(4):749–756. doi: 10.1161/01.res.62.4.749. [DOI] [PubMed] [Google Scholar]
  7. Haliday E. M., Ramesha C. S., Ringold G. TNF induces c-fos via a novel pathway requiring conversion of arachidonic acid to a lipoxygenase metabolite. EMBO J. 1991 Jan;10(1):109–115. doi: 10.1002/j.1460-2075.1991.tb07926.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hansbrough J. R., Takahashi Y., Ueda N., Yamamoto S., Holtzman M. J. Identification of a novel arachidonate 12-lipoxygenase in bovine tracheal epithelial cells distinct from leukocyte and platelet forms of the enzyme. J Biol Chem. 1990 Jan 25;265(3):1771–1776. [PubMed] [Google Scholar]
  9. Izumi T., Hoshiko S., Rådmark O., Samuelsson B. Cloning of the cDNA for human 12-lipoxygenase. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7477–7481. doi: 10.1073/pnas.87.19.7477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Maruyama T., Ueda N., Yoshimoto T., Yamamoto S., Komatsu N., Watanabe K. Immunohistochemical study of arachidonate 12-lipoxygenase in porcine tissues. J Histochem Cytochem. 1989 Jul;37(7):1125–1131. doi: 10.1177/37.7.2499621. [DOI] [PubMed] [Google Scholar]
  11. McClain D. A., Paterson A. J., Roos M. D., Wei X., Kudlow J. E. Glucose and glucosamine regulate growth factor gene expression in vascular smooth muscle cells. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8150–8154. doi: 10.1073/pnas.89.17.8150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nadler J. L., Natarajan R., Stern N. Specific action of the lipoxygenase pathway in mediating angiotensin II-induced aldosterone synthesis in isolated adrenal glomerulosa cells. J Clin Invest. 1987 Dec;80(6):1763–1769. doi: 10.1172/JCI113269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nakao J., Ooyama T., Ito H., Chang W. C., Murota S. Comparative effect of lipoxygenase products of arachidonic acid on rat aortic smooth muscle cell migration. Atherosclerosis. 1982 Sep;44(3):339–342. doi: 10.1016/0021-9150(82)90008-9. [DOI] [PubMed] [Google Scholar]
  14. Natarajan R., Dunn W. D., Stern N., Nadler J. Key role of diacylglycerol-mediated 12-lipoxygenase product formation in angiotensin II-induced aldosterone synthesis. Mol Cell Endocrinol. 1990 Aug 20;72(2):73–80. doi: 10.1016/0303-7207(90)90096-q. [DOI] [PubMed] [Google Scholar]
  15. Natarajan R., Gonzales N., Hornsby P. J., Nadler J. Mechanism of angiotensin II-induced proliferation in bovine adrenocortical cells. Endocrinology. 1992 Sep;131(3):1174–1180. doi: 10.1210/endo.131.3.1505459. [DOI] [PubMed] [Google Scholar]
  16. Natarajan R., Gonzales N., Xu L., Nadler J. L. Vascular smooth muscle cells exhibit increased growth in response to elevated glucose. Biochem Biophys Res Commun. 1992 Aug 31;187(1):552–560. doi: 10.1016/s0006-291x(05)81529-3. [DOI] [PubMed] [Google Scholar]
  17. Parthasarathy S., Wieland E., Steinberg D. A role for endothelial cell lipoxygenase in the oxidative modification of low density lipoprotein. Proc Natl Acad Sci U S A. 1989 Feb;86(3):1046–1050. doi: 10.1073/pnas.86.3.1046. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ruderman N. B., Williamson J. R., Brownlee M. Glucose and diabetic vascular disease. FASEB J. 1992 Aug;6(11):2905–2914. doi: 10.1096/fasebj.6.11.1644256. [DOI] [PubMed] [Google Scholar]
  19. Setty B. N., Graeber J. E., Stuart M. J. The mitogenic effect of 15- and 12-hydroxyeicosatetraenoic acid on endothelial cells may be mediated via diacylglycerol kinase inhibition. J Biol Chem. 1987 Dec 25;262(36):17613–17622. [PubMed] [Google Scholar]
  20. Shearman M. S., Naor Z., Sekiguchi K., Kishimoto A., Nishizuka Y. Selective activation of the gamma-subspecies of protein kinase C from bovine cerebellum by arachidonic acid and its lipoxygenase metabolites. FEBS Lett. 1989 Jan 30;243(2):177–182. doi: 10.1016/0014-5793(89)80125-5. [DOI] [PubMed] [Google Scholar]
  21. Sigal E., Craik C. S., Highland E., Grunberger D., Costello L. L., Dixon R. A., Nadel J. A. Molecular cloning and primary structure of human 15-lipoxygenase. Biochem Biophys Res Commun. 1988 Dec 15;157(2):457–464. doi: 10.1016/s0006-291x(88)80271-7. [DOI] [PubMed] [Google Scholar]
  22. Stern N., Golub M., Nozawa K., Berger M., Knoll E., Yanagawa N., Natarajan R., Nadler J. L., Tuck M. L. Selective inhibition of angiotensin II-mediated vasoconstriction by lipoxygenase blockade. Am J Physiol. 1989 Aug;257(2 Pt 2):H434–H443. doi: 10.1152/ajpheart.1989.257.2.H434. [DOI] [PubMed] [Google Scholar]
  23. Tso J. Y., Sun X. H., Kao T. H., Reece K. S., Wu R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acids Res. 1985 Apr 11;13(7):2485–2502. doi: 10.1093/nar/13.7.2485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wolf G., Neilson E. G., Goldfarb S., Ziyadeh F. N. The influence of glucose concentration on angiotensin II-induced hypertrophy of proximal tubular cells in culture. Biochem Biophys Res Commun. 1991 Apr 30;176(2):902–909. doi: 10.1016/s0006-291x(05)80271-2. [DOI] [PubMed] [Google Scholar]
  25. Ylä-Herttuala S., Rosenfeld M. E., Parthasarathy S., Glass C. K., Sigal E., Witztum J. L., Steinberg D. Colocalization of 15-lipoxygenase mRNA and protein with epitopes of oxidized low density lipoprotein in macrophage-rich areas of atherosclerotic lesions. Proc Natl Acad Sci U S A. 1990 Sep;87(18):6959–6963. doi: 10.1073/pnas.87.18.6959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yokoyama C., Shinjo F., Yoshimoto T., Yamamoto S., Oates J. A., Brash A. R. Arachidonate 12-lipoxygenase purified from porcine leukocytes by immunoaffinity chromatography and its reactivity with hydroperoxyeicosatetraenoic acids. J Biol Chem. 1986 Dec 15;261(35):16714–16721. [PubMed] [Google Scholar]
  27. Yoshimoto T., Miyamoto Y., Ochi K., Yamamoto S. Arachidonate 12-lipoxygenase of porcine leukocyte with activity for 5-hydroxyeicosatetraenoic acid. Biochim Biophys Acta. 1982 Dec 13;713(3):638–646. [PubMed] [Google Scholar]
  28. Yoshimoto T., Suzuki H., Yamamoto S., Takai T., Yokoyama C., Tanabe T. Cloning and sequence analysis of the cDNA for arachidonate 12-lipoxygenase of porcine leukocytes. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2142–2146. doi: 10.1073/pnas.87.6.2142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Yoshimoto T., Yamamoto Y., Arakawa T., Suzuki H., Yamamoto S., Yokoyama C., Tanabe T., Toh H. Molecular cloning and expression of human arachidonate 12-lipoxygenase. Biochem Biophys Res Commun. 1990 Nov 15;172(3):1230–1235. doi: 10.1016/0006-291x(90)91580-l. [DOI] [PubMed] [Google Scholar]
  30. Yu C. L., Tsai M. H., Stacey D. W. Serum stimulation of NIH 3T3 cells induces the production of lipids able to inhibit GTPase-activating protein activity. Mol Cell Biol. 1990 Dec;10(12):6683–6689. doi: 10.1128/mcb.10.12.6683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES