Subscribe to RSS
DOI: 10.1055/s-2005-871264
© Georg Thieme Verlag KG Stuttgart · New York
14-Deoxyandrographolide as a Platelet Activating Factor Antagonist in Bovine Neutrophils
Publication History
Received: September 14, 2004
Accepted: February 13, 2005
Publication Date:
18 July 2005 (online)

Abstract
14-Deoxyandrographolide (14-DAP) is a labdane diterpene isolated from Andrographis paniculata with previously reported calcium channel blocking activity. Its potential platelet activating factor (PAF) antagonistic activity in bovine neutrophils was assessed. 14-DAP, in concentrations between 10 - 100 μM, reduced the extracellular acidification rate and the intracellular alkalinization in a dose-dependent manner. In addition, 14-DAP reduced PAF-induced calcium flux in the presence of extracellular calcium, and tyrosine phosphorylation of a 44 kDa protein corresponding to the MAPKERK1. However, 14-DAP reduced the 3H-PAF binding with a Ki of 7.8 × 10 - 9 M, and a Hill slope of 0.63, suggesting that there is more than one binding site for 14-DAP. We concluded that 14-DAP is an effective antagonist of PAF-mediated processes in bovine neutrophils, probably by virtue of its calcium channel blocking property.
Key words
PAF - calcium - pH - neutrophils - 14-deoxyandrographolide - MAPK - Andrographis paniculata - Acanthaceae
References
- 1 Prescott S M, Zimmerman G A, Stafforini D M, McIntyre T M. Platelet-activating factor and related lipid mediators. Annu Rev Biochem. 2000; 69 419-45
- 2 Burgos R A, Hidalgo M A, Matthei S M, Hermosilla R, Folch H, Hancke J L. Determination of specific receptor sites for platelet activating factor in bovine neutrophils. Am J Vet Res. 2004; 65 628-36
- 3 Swain S D, Bunger P L, Sipes K M, Nelson L K, Jutila K L, Boylan S M, Quinn M T. Platelet-activating factor induces a concentration-dependent spectrum of functional responses in bovine neutrophils. J Leukocyte Biol. 1998; 64 817-27
- 4 Hidalgo M A, Ojeda F, Eyre P, LaBranche T P, Smith C, Hancke J L, Burgos R A. Platelet-activating factor increases pH(i) in bovine neutrophils through the PI3K-ERK1/2 pathway. Br J Pharmacol. 2004; 141 311-21
- 5 McClenahan D J, Evanson O A, Weiss D J. In vitro evaluation of the role of platelet-activating factor and interleukin-8 in Mannheimia haemolytica-induced bovine pulmonary endothelial cell injury. Am J Vet Res. 2002; 63 394-401
- 6 Montrucchio G, Alloatti G, Camussi G. Role of platelet-activating factor in cardiovascular pathophysiology. Physiol Rev. 2000; 80 1669-99
- 7 Negro Alvarez J M, Miralles Lopez J C, Ortiz Martinez J L, Abellan Aleman A, Rubio del Barrio R. Platelet-activating factor antagonists. Allergol Immunopathol (Madr). 1997; 25 249-58
- 8 Han B H, Yang H O, Kang Y H, Suh D Y, Go H J, Song W J, Kim Y C, Park M K. In vitro platelet-activating factor receptor binding inhibitory activity of pinusolide derivatives: a structure-activity study. J Med Chem. 1998; 41 2626-30
- 9 Amroyan E, Gabrielian E, Panossian A, Wikman G, Wagner H. Inhibitory effect of andrographolide from Andrographis paniculata on PAF-induced platelet aggregation. Phytomedicine. 1999; 6 27-31
- 10 Burgos R A, Loyola M, Hidalgo M A, Labranche T P, Hancke J L. Effect of 14-deoxyandrographolide on calcium-mediated rat uterine smooth muscle contractility. Phytother Res. 2003; 17 1011-5
- 11 Matsuda T, Kuroyanagi M, Sugiyama S, Umehara K, Ueno A, Nishi K. Cell differentiation-inducing diterpenes from Andrographis paniculata Nees. Chem Pharm Bull (Tokyo). 1994; 42 1216-25
- 12 Pick E. Microassays for superoxide and hydrogen peroxide production and nitroblue tetrazolium reduction using and enzyme immunoassays microplate reader. Methods Enzymol. 1986; 132 407-421
- 13 Trevani A S, Andonegui G, Giordano M, Lopez D H, Gamberale R, Minucci F, Geffner J R. Extracellular acidification induces human neutrophil activation. J Immunol. 1999; 162 4849-57
- 14 Osaki M, Sumimoto H, Takeshige K, Cragoe E J, Jr., Hori Y, Minakami S. Na+/H+ exchange modulates the production of leukotriene B4 by human neutrophils. Biochem J. 1989; 257 751-8
- 15 Simchowitz L. Intracellular pH modulates the generation of superoxide radicals by human neutrophils. J Clin Invest. 1985; 76 1079-89
- 16 Filep J G, Foldes-Filep E. Inhibition by calcium channel blockers of the binding of platelet-activating factor to human neutrophil granulocytes. Eur J Pharmacol. 1990; 190 67-73
- 17 Elzi D J, Hiester A A, Silliman C C. Receptor-mediated calcium entry is required for maximal effects of platelet activating factor primed responses in human neutrophils. Biochem Biophys Res Commun. 1997; 240 763-5
- 18 Hauser C J, Fekete Z, Adams J M, Garced M, Livingston D H, Deitch E A. PAF-mediated Ca2+ influx in human neutrophils occurs via store-operated mechanisms. J Leukocyte Biol. 2001; 69 63-8
- 19 Boucek M M, Snyderman R. Calcium influx requirement for human neutrophil chemotaxis: inhibition by lanthanum chloride. Science. 1976; 193 905-7
- 20 Granfeldt D, Samuelsson M, Karlsson A. Capacitative Ca2+ influx and activation of the neutrophil respiratory burst. Different regulation of plasma membrane- and granule-localized NADPH-oxidase. J Leukocyte Biol. 2002; 71 611-7
Dr. Rafael A. Burgos
Laboratory of Molecular Pharmacology
Institute of Pharmacology
Facultad de Ciencias Veterinarias
Universidad Austral de Chile
Valdivia
P.O. box 567
Chile
Email: rburgos1@uach.cl