Eicosanoid-mediated contractility of hepatic stellate cells

Abstract

To approach experimentally the problem of contractility, stellate cells from rats were isolated and grown on a flexible silicone rubber substrate. Increases or decreases in the number of wrinkles of the silicone membrane beneath the cells that were easily observable by microscopy was employed as semi-quantitative measure of stellate cell motility. Contraction of stellate cells accompanied by diminution of cell body size was induced by U46619 (a thromboxane A2 analogue) and prostaglandin (PG) F2 alpha. Wrinkle formation became detectable 1.5 min after addition of 2 microM-U46619 and reached its maximum 10-15 min later. The effect of PGF2 alpha was not so striking, but lasted for a longer period of time. On the other hand, dibutyryl cyclic AMP, Iloprost (a PGI2 analogue) and PGE2 led to the disappearance or decrease in the number of wrinkles, indicating relaxation of contracted stellate cells. For instance, after addition of 2 microM-Iloprost, 47, 75 and 82% of contracted stellate cells had relaxed within 5, 10 and 20 min respectively. Moreover, dibutyryl cyclic AMP induced disappearance of alpha-smooth muscle actin stress fibres. This response became recognizable 10 min after addition of dibutyryl cyclic AMP; 40 min later, 97% of stellate cells were devoid of stress fibres. Thus stellate cells are able to undergo reversible contraction in primary culture, and the contraction of these cells may be mediated by eicosanoids that can be produced within the liver.