Effects of vitamin E on the NF-kappaB pathway in rats treated with the peroxisome proliferator, ciprofibrate.

Peroxisome proliferators (PPs) are a diverse group of nongenotoxic compounds, which induce hepatic tumors in rodents. The mechanisms leading to hepatic tumors have not been elucidated, but oxidative stress may play a role in the process. Previous studies in our laboratory have shown that peroxisome proliferators activate the transcription factor nuclear factor-kappa B (NF-kappaB) and that this activation is mediated at least in part by oxidative stress. We therefore hypothesized that increased dietary vitamin E would decrease NF-kappaB DNA binding in rodents treated with ciprofibrate (CIP). In this study, 36 male Sprague-Dawley rats were fed a purified diet containing varying levels of vitamin E (10, 50, 250 ppm alpha-tocopherol acetate). After 28 days on the purified diet, seven animals per vitamin E group received 0.01% CIP in the diet for 10 days. Electrophoretic mobility shift assays (EMSAs) showed that CIP treatment increased DNA binding of NF-kappaB. Increased dietary alpha-tocopherol acetate inhibited CIP-induced NF-kappaB DNA binding. Because NF-kappaB translocates to the nucleus upon the phosphorylation and degradation of inhibitor of IkappaB, we also used Western blots to measure cytosolic protein levels of IkappaBalpha and IkappaBbeta, and the IkappaB kinases, IKKalpha and IKKbeta. IkappaBalpha protein levels were decreased in all three CIP-treated groups, with the 10 ppm vitamin E diet also decreasing IkappaBalpha levels in control rats. No difference in IkappaBbeta protein levels was observed among any of the groups. The CIP-treated rats generally had lower protein levels of IKKalpha and IKKbeta. This study supports our working hypothesis that an increased antioxidant environment can inhibit CIP-mediated NF-kappaB induction.