Platelet aggregation inhibitors from Philippine marine invertebrate samples screened in a new microplate assay

A new microplate assay for Ca(2+)-induced platelet aggregation as detected by Giemsa dye was used to screen marine invertebrate samples from the Philippines for inhibitors of human platelet aggregation. Out of 261 crude methanol extracts of marine sponges and tunicates, 25 inhibited aggregation at 2...

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Bibliographic Details
Published in:Marine biotechnology (New York, N.Y.) N.Y.), 2003-07, Vol.5 (4), p.395-400
Main Authors: Pimentel, Sheila Marie V, Bojo, Zenaida P, Roberto, Amy V D, Lazaro, Jose Enrico H, Mangalindan, Gina C, Florentino, Leila M, Lim-Navarro, Pilar, Tasdemir, Deniz, Ireland, Chris M, Concepcion, Gisela P
Format: Article
Language:English
Subjects:
Adenosine Diphosphate
Animals
Aplysina
Azure Stains
Bioassays
Blood Platelets - metabolism
Collagen
Epinephrine
Humans
Invertebrates
Invertebrates - chemistry
Macrocyclic Compounds
Marine
Marine biology
Marine invertebrates
Mass Spectrometry
Oxazoles - isolation & purification
Oxazoles - pharmacology
Pacific Ocean
Philippines
Platelet Aggregation - drug effects
Platelet Aggregation Inhibitors - isolation & purification
Platelet Aggregation Inhibitors - pharmacology
Porifera
Studies
Tunicata
Xestospongia
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Summary:A new microplate assay for Ca(2+)-induced platelet aggregation as detected by Giemsa dye was used to screen marine invertebrate samples from the Philippines for inhibitors of human platelet aggregation. Out of 261 crude methanol extracts of marine sponges and tunicates, 25 inhibited aggregation at 2 mg/ml. Inhibition of agonist-induced aggregation in an aggregometer was used to confirm results of the microplate assay and to determine the specific mode of inhibition of 2 samples. The marine sponge Xestospongia sp. yielded a xestospongin/araguspongine-type molecule that inhibited collagen-induced aggregation by 87% at 2 micro g/ml, and epinephrine-induced aggregation by 78% at 20 micro g/ml, while the marine sponge Aplysina sp. yielded 5,6-dibromotryptamine, which inhibited epinephrine-induced aggregation by 51% at 20 micro g/ml. In this study we have found that the microplate assay is a simple, inexpensive, yet useful preliminary tool to qualitatively screen a large number of marine samples for antiplatelet aggregation activity.
ISSN:1436-2228
1436-2236
DOI:10.1007/s10126-002-0080-3