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

Inflammation volume 12, pages 549–561 (1988)Cite this article

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.

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Affiliations

  1. Department of Biochemistry and Molecular Biophysics, Medical College of Virginia, Virginia Commonwealth University, Box 614, 23298-0614, Richmond, Virginia

    B. S. Vishwanath, A. A. Fawzy & R. C. Franson

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  1. B. S. Vishwanath
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  2. A. A. Fawzy
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Vishwanath, B.S., Fawzy, A.A. & Franson, R.C. Edema-inducing activity of phospholipase A2 purified from human synovial fluid and inhibition by aristolochic acid. Inflammation 12, 549–561 (1988). https://doi.org/10.1007/BF00914317

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Keywords

  • Substrate Concentration
  • Chemical Component
  • Human Platelet
  • Snake Venom
  • Aristolochic Acid