Abstract
Gossypol is present in antiinflammatory poultices made from the medicinal treeThespesia populnea. Isolated human neutrophils exposed to 3–20μM gossypol for 15–90 min were assayed in vitro for superoxide production and surface expression of Mac-1 (CD11b/CD18). Gossypol increased superoxide production in a time- and concentration-dependent fashion consistent with a moderate, delayed respiratory burst. Surface Mac-1 expression was increased within 15 min by 3–5μ M gossypol, resulting in a 14-fold increase over controls and a threefold greater increase over that produced by PMA. Staurosporine failed to block gossypol induction of superoxide and Mac-1, while EDTA inhibited induction of Mac-1 only, implicating a calcium-dependent mechanism. Gossypol increased intracellular calcium to peak levels, but1 in a delayed fashion as compared to FMLP. These findings demonstrate that gossypol is a highly potent stimulant of Mac-1 expression and suggest at least two protein kinase C-independent pathways of neutrophil activation. The resultant exhaustion of neutrophils may account for the antiinflammatory properties of plants containing gossypol.
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Jayaweera, D. M. A. 1982. Medicinal plants (indigenous and exotic) used in Ceylon, Part IV. A Publication of the National Science Council of Sri Lanka, Colombo.
King, J. J., andL. B. De Silva. 1968. Optically active gossypol fromThespesia populnea.Tetrahedron Lett. 3:261.
Datta, S. C., V. V. S. Mutri, andT. R. Seshadri. 1968. A new component of the flowers ofThespesia populnea: (+)-Gossypol.Curr. Sci. 37:135.
Abou, D. M. 1976. Physiological effects and metabolism of gossypol.Residue Rev. 61: 126–160.
Gombe, S. 1983. A review of the current status in male contraceptive studies.East Afr. Med. J. 60:203–211.
Benz, C., C. Hollander, M. Keniry, T. L. James, andM. Mitchell. 1987. Lactic dehydrogenase isozymes,31P magnetic resonance spectroscopy, and in vitro antimitochondrial tumor toxicity with gossypol and rhodamine-123.J. Clin. Invest. 79:517–523.
Benz, C. C., M. A. Keniry, J. M. Ford, A. J. Townsend, F. W. Cox, S Palayoor, S. A. Matlin, W. N. Hait, andK. H. Cowan. 1990. Biochemical correlates of the antitumor and antimitochondrial properties of gossypol enantiomers.Mol. Pharmacol. 37:840–847.
Flack, M. R., R. G. Pyle, N. M. Mullen, B. Lorenzo, Y. W. Wu, R. A. Knazek, B. C. Nisula, andM. M. Reidenberg. 1983. Oral gossypol in the treatment of metastatic adrenal cancer.J. Clin. Endocrinol. Metab. 76:1019–1024.
Frick, J., andN. R. Kalla. 1990. Clinical studies with gossypol. Current status.Acta Eur. Fertil. 21:99–100.
Abou, D. M., andJ. W. Dieckert. 1974. Gossypol: uncoupling of respiratory chain and oxidative phosphorylation.Life Sci. 14:1955–1963.
Bugeja, V., G. Charles, D. Collier, andD. Wilkie. 1988. Primary mitochondrial activity of gossypol in yeast and mammalian cells.Biochem. Pharmacol. 37:4217–4224.
Kim, I., andD. P. Waller. 1984. Specific inhibition of the testicular mitochondrial respiratory chain in vitro by gossypol.J. Androl. 5:424–430.
Benz, C. C., S. B. Iyer, H. S. Asgari, S. A. Matlin, F. R. Aronson, andA. Barchowsky. 1991. Gossypol effects on endothelial cells and tumor blood flow,Life Sci. 49:PL67-PL72.
Vedder, N. B., B. W. Fouty, R. K. Winn, J. M. Harlan, andC. L. Rice. 1989. Role of neutrophils in generalized reperfusion injury associated with resuscitation from shock.Surgery 106:509–516.
Vedder, N. B., R. K. Winn, C. L. Rice, E. Y. Chi, K. E. Arfors, andJ. M. Harlan. 1990. Inhibition of leukocyte adherence by anti-CD18 monoclonal antibody attenuates reperfusion injury in the rabbit ear.Proc. Natl. Acad. Sci. U. S. A. 87:2643–2646.
Babior, B. M.. 1984. The respiratory burst of phagocytes.J. Clin. Invest. 73:599–601.
Sbarra, A. J., andR. R. Strauss. 1988. The Respiratory Burst and Its Physiological Significance. Plenum Press, New York. 476 pp.
Berger, M., J. O'Shea, A. S. Cross, T. M. Folks, T. M. Chused, E. J. Brown, andM. M. Frank. 1984. Human neutrophils increase expression of Mac-1 as well as C3b receptors upon activation.J. Clin. Invest. 74:1566–1571.
Kishimoto, T. K., R. S. Larson, A. L. Corbi, M. L. Dustin, D. E. Staunton, andT. A. Springer. 1989. The leukocyte integrins.Adv. Immunol. 46:149–182.
Boyum, A. 1968. Isolation of mononuclear cells and granulocytes from human blood. Isolation of mononuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1g.Scand. J. Clin. Lab. Invest. 21(Suppl 97):77–89.
English, D., andB. R. Andersen. 1974. Single-step separation of red blood cells. Granulocytes and mononuclear leukocytes on a discontinuous density gradient of ficoll-hypaque.J. Immunol. Methods 5:249–252.
Azzi, A., C. Montecucco, andC. Richter. 1975. The use of acetylated ferricytochromec for the detection of superoxide radicals produced in biological membranes.Biochem. Biophys. Res. Commun. 65:597–603.
Butler, J., W. H. Koppenol, andE. Margoliash. 1982. Kinetics and mechanism of the reduction of ferricytochromec by the superoxide anion.J. Biol. Chem. 257:10747–10750.
Babior, B. M., R. S. Kipnes, andJ. T. Curnutte. 1973. Biological defense mechanisms. The production by leukocytes of superoxide, a potential bactericidal agent.J. Clin. Invest. 52:741–747.
Metcalf, J. A., J. I. Gallin, W. M. Nauseef, andR. K. Root. 1986. Laboratory Manual of Neutrophil Function. Raven, New York. 191 pp.
English, D., andV. Graves. 1992. Simultaneous mobilization of Mac-1 (CD11b/CD18) and formyl peptide chemoattractant receptors in human neutrophils.Blood 80:776–787.
Sasaki, D. T., S. E. Dumas, andE. G. Engleman. 1987. Discrimination of viable and nonviable cells using propidium iodide in two color immunofluorescence.Cytometry 8:413–420.
Minta, A., J. P. Kao, andR. Y. Tsien. 1989. Fluorescent indicators for cytosolic calcium based on rhodamine and fluorescein chromophores.J. Biol. Chem. 264:8171–8178.
Sei, Y., andP. K. Arora. 1991. Quantitative analysis of calcium (CA2+) mobilization after stimulation with mitogens or anti-CD3 antibodies. Simultaneous fluo-3 and immunofluorescence flow cytometry.J. Immunol. Methods 137:237–244.
Tsien, R. Y. 1988. Fluorescence measurement and photochemical manipulation of cytosolic free calcium.Trends Neurosci. 11:419–424.
Zar, J. H. 1984. Biostatistical Analysis. Prentice-Hall, Englewood Cliffs, New Jersey. 620 pp.
Barchowsky, A., K. Tabrizi, R. S. Kent, andA. R. Whorton. 1989. Inhibition of prostaglandin synthesis after metabolism of menadione by cultured porcine endothelial cells.J. Clin. Invest. 83:1153–1159.
Wang, N. G., M. Z. Guan, andH. P. Lei. 1984. Effects of (−) and (+) gossypol in fertility of male rats.Acta Pharm. Sinica 19:932–934.
Schwartz, B. R., andJ. M. Harlan. 1989. Sulfhydryl reducing agents promote neutrophil adherence without increasing surface expression of CD11b/CD18 (Mac-1, Mol).Biochem. Biophys. Res. Commun. 165:51–57.
Vedder, N. B., andJ. M. Harlan. 1988. Increased surface expression of CD11b/CD18 (Mac-1) is not required for stimulated neutrophil adherence to cultured endothelium.J. Clin. Invest. 81:676–682.
Wymann, M. P., V. von Tscharner, D. A. Deranleau, andM. Baggiolini. 1987. The onset of the respiratory burst in human neutrophils. Real-time studies of H2O2 formation reveal a rapid agonist-induced transduction process.J. Biol. Chem. 262:12048–12053.
Sugioka, K., M. Nakano, H. Totsune-Nakano, H. Minakami, S. Tero-Kubota, andY. Ikegami. 1988. Mechanism of O −2 generation in reduction and oxidation cycle of ubiquinones in a model of mitochondrial electron transport systems.Biochim. Biophys. Acta 936:377–385.
Keniry, M. A., C. Hollander, andC. C. Benz. 1989. The effect of gossypol and 6-aminonicotinamide on tumor cell metabolism: A31P-magnetic resonance spectroscopic study.Biochem. Biophys. Res. Commun. 164:947–953.
Tanphaichitr, N., L. B. Chen, andA. R. Bellve. 1984. Direct effect of gossypol on TRST cells: perturbation of rhodamine 123 accumulation in mitochondria.Biol. Reprod. 31:1049–1060.
Ogle, J. D., G. Noel, R. M. Sramkoski, C. K. Ogle, andJ. W. Alexander. 1990. Effects of chemotactic peptide f-MET-LEU-PHE (FMLP) on C3b receptor (CR1) expression and phagocytosis of microspheres by human neutrophils.Inflammation 14:337–353.
Tso, W. W., andC. S. Lee. 1982. Gossypol uncoupling of respiratory chain and oxidative phosphorylation in ejaculated boar spermatozoa.Contraception 25:649–655.
Tuszynski, G. P., andG. Cossu. 1984. Differential cytotoxic effect of gossypol on human melanoma, colon carcinoma, and other tissue culture cell lines.Cancer Res. 44:768–771.
Lee, C. Y. G., Y. S. Moon, J. H. Yuan, andA. F. Chen. 1982. Enzyme inactivation and inhibition by gossypol.Mol. Cell. Biochem. 47:65–70.
Tahir, M. K., C. Guthenberg, andB. Mannervik. 1985. Inhibitors for distinction of three types of human glutathione transferase.FEBS Lett. 181:249–252.
Kimura, K., K. Sakurada, andN. Katoh. 1985. Inhibition by gossypol of phospholipidsensitive Ca2+-dependent protein kinase from pig testis.Biochim. Biophys. Acta 839:276–280.
Olgiati, K. L., D. G. Toscano, W. M. Atkins, andW. J. Toscano. 1984. Gossypol inhibition of adenylate cyclase.Arch. Biochem. Biophys. 231:411–415.
Rosenberg, L. J., R. C. Adlakha, D. M. Desai, andP. N. Rao. 1986. Inhibition of DNA polymerase alpha by gossypol.Biochim. Biophys. Acta 866(4):258–267.
Grankvist, K. 1989. Gossypol-induced free radical toxicity to isolated islet cells.Int. J. Biochem. 21:853–856.
Laughton, M. J., B. Halliwell, P. J. Evans, andJ. R. Hoult. 1989. Antioxidant and prooxidant actions of the plant phenolics quercetin, gossypol and myricetin. Effects on lipid peroxidation, hydroxyl radical generation and bleomycin-dependent damage to DNA.Biochem. Pharmacol. 38:2859–2865.
Zaidi, R., andS. M. Hadi. 1992. Strand scission in DNA by gossypol and Cu(II): role of Cu(I) and oxygen-free radicals.J. Biochem. Toxicol. 7:213–217.
de Peyster A., A. Quintanilha, L. Packer, andM. T. Smith. 1984. Oxygen radical formation induced by gossypol in rat liver microsomes and human sperm.Biochem. Biophys. Res. Commun. 118:573–579.
Wu, D. F., andY. W. Yu. 1986. Superoxide free radical formation stimulated by (+/−), (+), (−) gossypol in rat liver and kidney microsomes.Proc. Chin. Acad. Med. Sci. Peking Union Med. Coll. 1:150–156.
Moelleken, B. R., S. J. Mathes, A. Amerhauser, H. Scheuenstuhl, andT. K. Hunt. 1991. An adverse wound environment activates leukocytes prematurely.Arch. Surg. 126:225–230.
Eshima, I., S. J. Mathes, andP. Paty. 1990. Comparison of the intracellular bacterial killing activity of leukocytes in musculocutaneous and random-pattern flaps.Plast. Reconstr. Surg. 86:541–547.
Fitzpatrick, L. R., J. S. Bostwick, M. Renzetti, R. G. Pendleton, andD. L. Decktor. 1990. Antiinflammatory effects of various drugs on acetic acid induced colitis in the rat.Agents Actions 30:393–402.
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Benhaim, P., Mathes, S.J., Hunt, T.K. et al. Induction of neutrophil Mac-1 integrin expression and superoxide production by the medicinal plant extract gossypol. Inflammation 18, 443–458 (1994). https://doi.org/10.1007/BF01560692
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DOI: https://doi.org/10.1007/BF01560692