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(Hypertension. 1998;31:206.)
© 1998 American Heart Association, Inc.

Scientific Contributions

Methacholine-Induced Contraction of Rabbit Pulmonary Artery: Role of Platelet-Endothelial Transcellular Thromboxane Synthesis

Sandra L. Pfister; David D. Deinhart; William B. Campbell

From the Departments of Pharmacology and Toxicology and Internal Medicine, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226.

Correspondence to Sandra L. Pfister, PhD, Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226. E-mail spfister{at}post.its.mcw.edu

Arachidonic acid- and methacholine-induced contractions of rabbit pulmonary arteries are mediated by thromboxane (TX) A₂. Although removal of the endothelium abolishes the contractions, endothelial cells isolated from pulmonary arteries do not synthesize TXA₂. Further studies described here showed that the expression of TX synthase was evident in platelets and intact pulmonary artery but not in endothelial cells. These studies examined the role of platelet TXA₂ production in the vasoconstrictor response to methacholine. Endothelial cells were incubated with platelets in the presence or absence of methacholine. Methacholine caused an increase in TXB₂ production. Pretreatment of endothelial cells with aspirin (100 µmol/L) before the addition of platelets did not impair the ability of methacholine to increase TXB₂ synthesis. Conversely, if platelets were pretreated with aspirin, methacholine failed to stimulate TXB₂. Using endothelial cells with their cellular lipids labeled with [³H]arachidonic acid, methacholine did not stimulate the production of [³H]TXB₂. When the endothelial cells were incubated with methacholine and control platelets, [³H]TXB₂ was detected. If aspirin-treated platelets were incubated with endothelial cells, methacholine did not increase the production of [³H]TXB₂. However, pretreatment of the endothelial cells with aspirin did not affect the ability of methacholine to induce [³H]TXB₂ release. This suggests that methacholine stimulated the endothelial cell to release arachidonic acid, which was transferred to the platelets and metabolized to TXA₂. To test whether this cell-cell interaction is necessary for methacholine-induced contractions, rabbits were administered aspirin (20 mg/kg) for 2 days. On day 4, methacholine-induced contractions of pulmonary arteries were depressed in aspirin-treated compared with control subjects. Control arteries synthesized 6-keto-prostaglandin F₁ and TXB₂. Aspirin treatment inhibited both pulmonary artery and platelet TXB₂ production but had no effect on vessel 6-keto-prostaglandin F₁. These studies implicate platelets as a vascular source of TXA₂ and indicate that both endothelial cells and platelets may be required for methacholine-induced TXA₂ synthesis and vasoconstriction.

Key Words: thromboxane A₂ • cyclooxygenase • platelets • arachidonic acid • endothelial cells • endothelium-derived contracting factor

Abbreviations: PG = prostaglandin • PMN = polymorphonuclear leukocyte • RIA = radioimmunoassay • RP-HPLC = reverse-phase high-pressure liquid chromatography • TX = thromboxane

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