Advertisement

Inflammation

, Volume 12, Issue 6, pp 549–561 | Cite as

Edema-inducing activity of phospholipase A2 purified from human synovial fluid and inhibition by aristolochic acid

  • B. S. Vishwanath
  • A. A. Fawzy
  • R. C. Franson
Original Articles

Abstract

A neutral-active, Ca2+-dependent phospholipase A2 (PLA2) purified 11,000-fold from human synovial fluid (HSF) induced edema when injected into the mouse foot pad. The edema produced by HSF-PLA2 was dose-dependent and was positively correlated with the dose-dependent in vitro expression of PLA2 activity. Maximum edema was achieved within 45 min after the injection and persisted for atleast 6 h. Aristolochic acid [8-methoxy-6-nitrophenanthro(3,4-d)-1,3-dioxole-5-carboxylic acid], a major chemical component derived from various species ofAristolochia plant, produced a dose-dependent inhibition of in vitro phospholipid hydrolysis by HSF-PLA2, porcine pancreatic PLA2, snake venom (Naja naja) PLA2, and PLA2 isolated from human platelet. The sensitivity of these PLA2s to inhibition by aristolochic acid varied markedly: HSF-PLA2>N.naja PLA2>human platelet PLA2>porcine pancreatic PLA2. The inhibition of HSF-PLA2 by aristolochic acid was independent of substrate concentration (18–144ΜM and Ca2+ concentration (0.1–4.0 mM). These observations indicate that inhibition of HSF-PLA2 by aristolochic acid may result from direct interaction with the enzyme. When aristolochic acid was mixed with HSF-PLA2 and then injected into the mouse foot pad, edema was inhibited in a dose-dependent manner and was positively correlated with in vitro inhibition of PLA2 activity. Alkylation of HSF-PLA2 withp-bromophenacyl bromide concomitantly inhibited both enzyme and edema-inducing activity. These results clearly demonstrate that the neutral-active, Ca2+-dependent PLA2 isolated from human synovial fluid is proinflammatory and that catalytic activity is positively correlated with in vivo proinflammatory effects.

Keywords

Substrate Concentration Chemical Component Human Platelet Snake Venom Aristolochic Acid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Van Deenen, L. L. M., andG. H. de Haas. 1963. The substrate specificity of phospholipase A.Biochim. Biophys. Acta 70:538–553.Google Scholar
  2. 2.
    Van Den Bosch, H., andL. L. M. Van Deenen. 1965. Chemical structure and biochemical significance of lysolecithins from rat liver.Biochim. Biophys. Acta 106:326–337.Google Scholar
  3. 3.
    Franson, R. C. 1981. Intracellular metabolism of ingested phospholipids.In Liposomes from Physical Structure to Therapeutic Applications. G. Knight, editor. Elsevier-North Holland, New York. 349–380.Google Scholar
  4. 4.
    Waite, M., H. Griffin, andR. C. Franson. 1976. The phospholipase A of lysosomes.In Lysosomes in Biology and Pathology, Vol. 5. A. Neuberger and E. L. Tatus, editors. Elsevier-North Holland, New York. 257–305.Google Scholar
  5. 5.
    Vadas, P., andJ. B. Hay. 1980. The release of phospholipase A2 from aggregated platelets and stimulated macrophages of sheep.Life Sci. 26:1721–1729.Google Scholar
  6. 6.
    Traynor, J. R., andK. S. Authi. 1981. Phospholipase A2 activity of lysosomal origin secreted by polymorphonuclear leucocytes during phagocytosis or on treatment with calcium.Biochim. Biophys. Acta 655:571–577.Google Scholar
  7. 7.
    Wightman, P. D., M. E. Dahlgren, P. Davies, andR. J. Bonney. 1981. The selective release of phospholipase A2 by resident mouse peritoneal macrophages.Biochem. J. 200:441–444.Google Scholar
  8. 8.
    Vadas, P., andJ. B. Hay. 1982. The appearance and significance of phospholipase A2 in lymph draining tuberculin reactions.Am. J. Pathol. 107:285–291.Google Scholar
  9. 9.
    Lanni, C., andE. L. Becker. 1983. Release of phospholipase A2 activity from rabbit peritoneal neutrophils by f-Met-Leu-Phe.Am. J. Pathol. 113:90–94.Google Scholar
  10. 10.
    Horigome, K., M. Hayakawa, K. Inoue, andS. Nojima. 1987. Selective release of phospholipase A2 and lysophosphatidylserine-specific lysophospholipase from rat platelet.J. Biochem. 101:53–61.Google Scholar
  11. 11.
    Nevalainen, T. J. 1980. The role of phospholipase A in acute pancreatitis.Scand. J. Gastroenterol. 15:641–650.Google Scholar
  12. 12.
    Vadas, P. 1984. Plasma phospholipase A2 levels correlate with the hemodynamic and pulmonary changes in gram negative septic shock in man.J. Lab. Clin. Med. 104:873–881.Google Scholar
  13. 13.
    Pruzanski, W., P. Vadas, E. Stefanski, andM. B. Urowitz. 1985. Phospholipase A2 activity in sera and synovial fluids in rheumatoid arthritis and osteoarthritis. Its possible role as a proinflammatory enzyme.J. Rheumatol. 12:211–216.Google Scholar
  14. 14.
    Vadas, P., andW. Pruzanski. 1984. Role of extracellular phospholipase A2 in inflammation.Adv. Inflam. Res. 7:51–59.Google Scholar
  15. 15.
    Chilton, F. H., J. T. O'Flaherty, C. E. Walsh, M. J. Thomas, R. L. Wykle, L. R. Dechatelet, andB. M. Waite. 1982. Platelet activating factor. Stimulation of the lipoxygenase pathway in polymorphonuclear leukocytes by 1-O-alkyI-2-O-acetyl-sn-glycero-3-phosphocholine.J. Biol. Chem. 257:5402–5407.Google Scholar
  16. 16.
    Pruzanski, W., P. Vadas, andV. Fornasier. 1986. Inflammatory effect of intradermal administration of soluble phospholipase A2 in rabbits.J. Invest. Dermatol. 86:380–383.Google Scholar
  17. 17.
    Marshall, L. A., E. Blazek, andJ. Chang. 1987. Characterization of an in vivo model for evaluation of phospholipase A2 inhibitors.Fed. Proc. 46:854.Google Scholar
  18. 18.
    Vishwanath, B. S., R. M. Kini, andT. V. Gowda. 1987. Characterization of three edemainducing phospholipase A2 enzymes from habu (Trimeresurus flavoviridis) venom and their interaction with the alkaloid aristolochic acid.Toxicon 25:501–515.Google Scholar
  19. 19.
    Vishwanath, B. S., andT. V. Gowda. 1987. Interaction of aristolochic acid withVipera russelli phospholipase A2: Its effect on enzymatic and pathological activities.Toxicon 25:929–937.Google Scholar
  20. 20.
    Tsai, L. H., L. L. Yang, andC. Chang. 1980. Inactivation of Formosan snake venoms in vivo by aristolochic acid, the chemical component ofAristolochia radix.Tai-wan K'o Hsueh. 34:40–44.Google Scholar
  21. 21.
    Fawzy, A. A., R. Dobrow, andR. C. Franson. 1986. Modulation of human synovial phospholipase A2 by monovalent cations.Fed. Proc. 45:1560.Google Scholar
  22. 22.
    Raghupathi, R., andR. C. Franson. 1987. Isolation of a Ca2+-dependent phospholipase A2 from human platelets.Fed. Proc. 46:1945.Google Scholar
  23. 23.
    Franson, R., P. Patriarca, andP. Elsbach. 1974. Phospholipid metabolism by phagocytic cells. Phospholipase A2 associated with rabbit polymorphonuclear leukocyte granules.J. Lipid Res. 15:380–388.Google Scholar
  24. 24.
    Yamakawa, M., M. Nozaki, andZ. Hokama. 1976. Fractionation of Sakishima-habu (Trimeresurus elegans) venom and lethal hemorrhagic and edema forming activity of the fractions.In Animal Plant and Microbial Toxins, Vol. 1. A. Ohsaka, K. Hayashi, and Y. Sawai, editors. Plenum, New York. 97–109.Google Scholar
  25. 25.
    Franson, R. C., andD. Weir. 1983. Inhibition of a potent phospholipase A2 activity by nonsteroidal antiinflammatory agents in synovial fluid from patients with rheumatoid arthritis.Clin. Res. 31:650A.Google Scholar
  26. 26.
    Robinson, R. C., D. P. Curran, andP. J. Hamer. 1982. Prostaglandins and related compounds in inflammatory rheumatic disease.In Advances in Inflammation Research, vol. 3. M. Ziff, G. P. Velo, and S. Gorini, editors. Raven Press, New York, 17–27.Google Scholar
  27. 27.
    Hirata, F., E. Schiffmann, K. Venkatasubramanian, D. Salomon, andJ. Axelrod. 1980. A phospholipase A2 inhibitory protein in rabbit neutrophils induced by glucocorticoids.Proc. Natl. Acad. Sci. U.S.A. 77:2533–2536.Google Scholar
  28. 28.
    Blackwell, G. J., R. Carnuccio, M. Dirosa, R. J. Flower, L. Parente, andP. Persico. 1980. Macrocortin: A polypeptide causing the anti-phospholipase effect of glucocorticoids.Nature 287:147–149.Google Scholar
  29. 29.
    Cloix, J. F., O. Colard, B. Rothhut, andF. Russo-Marie. 1983. Characterization and partial purification of “renocortins”: Two polypeptides formed in renal cells causing the anti-phospholipase like action of glucocorticoids.Br. J. Pharmacol. 79:313–321.Google Scholar
  30. 30.
    Davidson, F. F., E. A. Dennis, M. Powell, andJ. R. Glenney. 1987. Inhibition of phospholipase A2 by “lipocortins” and calpactins. An effect of binding to substrate phospholipids.J. Biol. Chem. 262:1698–1705.Google Scholar
  31. 31.
    Jesse, R., andR. Franson. 1979. Modulation of purified phospholipase A2 activity from human platelets by calcium and indomethacin.Biochim. Biophys. Acta 575:467–470.Google Scholar
  32. 32.
    Franson, R., D. Eisen, R. Jesse, andC. Lanni. 1980. Inhibition of highly purified mammalian phospholipase A2 by nonsteroidal anti-inflammatory agents: Modulation by calcium ions.Biochem. J. 865:633–636.Google Scholar
  33. 33.
    Condrea, E., J. E. Fletcher, B. E. Rapuano, C.-C. Yang, andP. Rosenberg. 1981. Dissociation of enzymatic activity from lethality and pharmacological properties by carbamylation of lysines inNaja nigricollis, andNaja naja atra snake venom phospholipase A2.Toxicon 19:705–720.Google Scholar
  34. 34.
    Boffa, M. C., C. Rothen, H. M. Verheij, R. Verger, andG. H. de Haas. 1981. Classification of phospholipase A2 based upon their anticoagulant activity and penetration ability into phospholipid monolayers.In Natural Toxins. D. Eaker, and T. Wadstrom, editors. Pergamon Press, Oxford. 131–138.Google Scholar
  35. 35.
    Di Rosa, M., J. P. Giroud, andD. A. Willoughby. 1971. Studies of the mediators of the acute inflammatory response induced in rats in different sites by carrageenan and turpentine.J. Pathol. 104:15–29.Google Scholar
  36. 36.
    Bekemeier, H., A. J. Giessler, R. Hirschelmann, andG. Metzner. 1979. Relationship between prostaglandins and lysophosphatides in the inflammatory process.Agents Actions (Suppl. 4): 286–292.Google Scholar
  37. 37.
    Weltzien, H. U., 1979. Cytolytic and membrane-perturbing properties of lysophosphatidylcholine.Biochim. Biophys. Acta 559:259–287.Google Scholar
  38. 38.
    Vishwanath, B. S., A. G. A. Rao, andT. V. Gowda. 1987. Interaction of phospholipase A2 fromVipera russelli venom with aristolochic acid: A circular dichroism study.Toxicon 25:939–946.Google Scholar
  39. 39.
    Yaron, M. 1982. Structure and function of connective tissue joints, and synovial fluid.In Principles of Rheumatic Diseases. R. S. Panush, editor. John Wiley & Sons, New York. 3–14.Google Scholar
  40. 40.
    Kyger, E. M., andR. C. Franson. 1984. Non-specific inhibition of enzymes byp-bromophenacyl bromide. Inhibition of human platelet phospholipase C and modification of sulfhydryl groups.Biochim. Biophys. Acta 794:96–103.Google Scholar
  41. 41.
    Volwerk, J. J., W. A. Pieterson, andG. H. De Haas. 1974. Histidine at the active site of phospholipase A2.Biochemistry 13:1446–1454.Google Scholar
  42. 42.
    Condrea, E., J. E. Fletcher, B. E. Rapuano, C.-C. Yang, andP. Rosenberg. 1981. Effect of modification of one histidine residue on the enzymatic and pharmacological properties of a toxic phospholipase A2 fromNaja nigricollis snake venom and less toxic phospholipase A2 fromHematchatus haemachatus andNaja naja atra snake venoms.Toxicon 19:61–71.Google Scholar
  43. 43.
    Dennis, E. A., 1983. Phospholipases.In The Enzymes, Vol. 16. P. D. Boyer, editor. Academic Press, New York. 307–353.Google Scholar
  44. 44.
    Kini, R. M., andS. Iwanaga. 1986. Structure-function relationships of phospholipases I: Prediction of presynaptic neurotoxicity.Toxicon 24:527–541.Google Scholar
  45. 45.
    Kini, R. M., andS. Iwanaga. 1986. Structure-function relationships of phospholipases II: Charge density distribution and the myotoxicity of presynaptically neurotoxic phospholipases.Toxicon 24:895–905.Google Scholar
  46. 46.
    Rosenberg, P., E. Condrea, B. Rapuano, K. Soons, andC.-C. Yang. 1983. Dissociation of pharmacological and enzymatic activities of snake venom phospholipase A2 by modification of carboxylate groups.Biochem. Pharmacol. 32:3525–3530.Google Scholar
  47. 47.
    Barrington, P. L., E. Condrea, K. R. Soons, C.-C. Yang, andP. Rosenberg. 1984. Effect of carboxylate group modification on enzymatic and cardiotoxic properties of snake venom phospholipase A2.Toxicon 22:743–758.Google Scholar
  48. 48.
    Harris, J. B., andC. A. MacDonell. 1981. Phospholipase A2 activity of notexin and its role in muscle damage.Toxicon 19:419–430.Google Scholar
  49. 49.
    Verheij, H. M., M. C. Boffa, C. Rothen, M. C. Bryckaert, R. Verger, andG. H. De Haas. 1980. Correlation of enzymatic activity and antocoagulant properties of phospholipase A2.Eur. J. Biochem. 112:25–32.Google Scholar
  50. 50.
    Fraenkel-Conrat, H. 1983. Snake venom neurotoxins related to phospholipase A2.J. Toxicol-Toxin Rev. 1:205–221.Google Scholar
  51. 51.
    Huang, H.-C. 1984. Release of slow reacting substance from the guinea-pig lung by phospholipase A2 ofVipera russelli snake venom.Toxicon 22:359–372.Google Scholar
  52. 52.
    Vadas, P., andJ. B. Hay. 1983. Involvement of circulating phospholipase A2 in the pathogenesis of the hemodynamic changes in endotoxic shock.Can. J. Physiol. Pharmacol. 61:561–566.Google Scholar

Copyright information

© Plenum Publishing Corporation 1988

Authors and Affiliations

  • B. S. Vishwanath
    • 1
  • A. A. Fawzy
    • 1
  • R. C. Franson
    • 1
  1. 1.Department of Biochemistry and Molecular BiophysicsMedical College of Virginia, Virginia Commonwealth UniversityRichmond

Personalised recommendations