Interleukin 1 beta and interleukin 6 repress clofibric acid induction of different P450 isoforms in cultured foetal rat hepatocytes.

Abstract

1. Expression of various P450 subfamilies (1A, 2A, 2B, 2C, 3A) have been studied in cultured foetal rat hepatocytes after treatment with clofibric acid, a peroxisome proliferator and prototypic CYP4A inducer in vitro. Ethoxyresorufin O-deethylase activity (EROD, a CYP1A-related activity) as well as 7 alpha-, 16 alpha-, 2 alpha- and 6 beta-testosterone hydroxylase activities (CYP2A, 2B, 2C11 and 3A respectively) were determined during culture. Levels of the corresponding P450 apoproteins were measured by Western blotting. 2. Clofibric acid was able to induce all the P450-dependent activities studied. In most cases this induction required the additional presence of dexamethasone, an agent which promotes differentiation and favours long-term maintenance of the hepatocytes. 3. The major pro-inflammatory cytokines, IL-1 beta and IL-6, decrease the levels of the clofibric acid-induced P450 isoforms, except CYP1A, which was insensitive to IL-6, previous studies having shown that IL-1 beta represses lauric acid 12-hydroxylase activity after induction by clofibric acid. The effects of these cytokines were clearly dose- and time-dependent. The decrease in enzyme activity correlated with a decrease in apoprotein content. 4. The ability of clofibric acid to induce P450 isoforms highlights the complexity of P450 regulation by peroxisome proliferators. Our results confirm, moreover, that different P450 subfamilies are differentially affected by IL-1 beta and IL-6.