Effects of Trypsin on Cytosolic Calcium Levels in Rat Aortic Endothelium

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Effect of trypsin on vascular tone and cytosolic calcium concentration ([Ca2+]i) of endothelial and smooth muscle cells were examined in rat aorta. [Ca2+]i was measured by a calcium indicator, fura-PE3, simultaneously with vascular tone. In endothelium-intact rat aorta, carbachol and trypsin increased [Ca2+]i in a dose-dependent manner. In endothelium-denuded rat aorta, carbachol did not change [Ca2+]i, but trypsin slightly increased [Ca2+]i. Addition of trypsin to the norepinephrine-stimulated rat aorta relaxed the muscle with additional increase in [Ca2+]i. In calcium-free condition, trypsin induced a transient increase in [Ca2+]i. Trypsin-induced endothelium-dependent relaxation was inhibited by the preincubation with l-NMMA, an endothelial NO synthase inhibitor, U-73122, a phospholipase C inhibitor, cyclopiazonic acid, a sarcoplasmic/endoplasmic reticulum Ca2+-ATPase blocker, and lanthanum, a non-selective Ca2+ channel blocker. However, indomethacin, a non-selective cyclooxygenase inhibitor, and SKF-96365, a store-operated Ca2+-channel blocker, had no effect on the trypsin-induced relaxation. These results suggest that trypsin increases [Ca2+]i in the endothelial cells through SKF-96365-insensitive Ca2+ channels, and regulates the release of NO which resulted in the relaxation of rat aorta.

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