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Am J Physiol Cell Physiol 281: C1038-C1045, 2001;
0363-6143/01 $5.00
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Vol. 281, Issue 3, C1038-C1045, September 2001

Regulation of PGE2 and PGI2 release from human umbilical vein endothelial cells by actin cytoskeleton

Sara J. Sawyer, Suzanne M. Norvell, Suzanne M. Ponik, and Fredrick M. Pavalko

Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, Indiana 46202

Disruption of microfilaments in human umbilical vein endothelial cells (HUVEC) with cytochalasin D (cytD) or latrunculin A (latA) resulted in a 3.3- to 5.7-fold increase in total synthesis of prostaglandin E2 (PGE2) and a 3.4- to 6.5-fold increase in prostacyclin (PGI2) compared with control cells. Disruption of the microtubule network with nocodazole or colchicine increased synthesis of PGE2 1.7- to 1.9-fold and PGI2 1.9- to 2.0-fold compared with control cells. Interestingly, however, increased release of PGE2 and PGI2 from HUVEC into the media occurred only when microfilaments were disrupted. CytD treatment resulted in 6.7-fold more PGE2 and 3.8-fold more PGI2 released from HUVEC compared with control cells; latA treatment resulted in 17.7-fold more PGE2 and 11.2-fold more PGI2 released compared with control cells. Both increased synthesis and release of prostaglandins in response to all drug treatments were completely inhibited by NS-398, a specific inhibitor of cyclooxygenase-2 (COX-2). Disruption of either microfilaments using cytD or latA or of microtubules using nocodazole or colchicine resulted in a significant increase in COX-2 protein levels, suggesting that the increased synthesis of prostaglandins in response to drug treatments may result from increased activity of COX-2. These results, together with studies demonstrating a vasoprotective role for prostaglandins, suggest that the cytoskeleton plays an important role in maintenance of endothelial barrier function by regulating prostaglandin synthesis and release from HUVEC.

cyclooxygenase; microfilaments; microtubules; vasoprotection; prostaglandin E2; prostacyclin


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