RESUMEN
Cholesterol metabolized to 7alpha-hydroxylated bile acids is a principle pathway of cholesterol degradation. Cholesterol 7alpha-hydroxylase (CYP7A1) is the initial and rate-determining enzyme in the "classic pathway" of bile acid synthesis. An "alternative" pathway of bile acid synthesis begins with 27-hydroxylation of cholesterol by 27-hydroxylase (CYP27), followed by 7alpha-hydroxylation by oxysterol 7alpha-hydroxylase (CYP7B1). The aim of the current study was to investigate the regulation of CYP7B1 by bile acids, cholesterol, and thyroid hormone in a previously well-studied in vivo model of bile acid synthesis, and to compare its regulation to that of CYP7A1. Three study groups were examined. In the first, male Sprague-Dawley rats with intact enterohepatic circulations were fed normal chow (controls), cholestyramine (CT), cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), or cholesterol (Chol). In the second group, taurocholate (TCA) was continuously intraduodenally infused for 48 hours to chronic biliary diverted rats. In a third set of studies, squalestatin, an inhibitor of cholesterol synthesis, was intravenously infused for 48 hours. In a fourth set of studies, the diurnal variation in CYP7B1 was compared to that of CYP7A1. At the end of each study livers were harvested, and CYP7B1 and CYP7A1 activities and mRNA levels were determined. Complete biliary diversion significantly increased the specific activity (SA) of both CYP7B1 ( upward arrow 212%; P <.002) and CYP7A1 ( upward arrow 212%; P <.007). Intraduodenal infusion of TCA to rats with biliary diversion decreased SA of both CYP7B1 ( downward arrow 29%; P <.001) and CYP7A1 ( downward arrow 46%; P <.01). The addition of CA, CDCA, or DCA to rat chow led to downregulation of CYP7B1 SAs by 42% (P <.003), 51% (P <.009), and 47% (P <.003), and CYP7A1 SAs by 32% +/- 6% (P <.003), 73% +/- 9% (P <.002), and 60% +/- 13% (P <.004), respectively. CT feeding upregulated both CYP7B1 ( upward arrow 136%; P <.004) and CYP7A1 ( upward arrow 216%; P <.001) SAs. While Chol feeding significantly upregulated CYP7A1 SA, no significant increase in CYP7B1 SA was found. Conversely, as previously shown in vitro, inhibition of cholesterol synthesis significantly suppressed both CYP7A1 and CYP7B1 activity and mRNA levels. Both CYP7B1 and CYP7A1 underwent diurnal variation, with peak and trough values for CYP7B1 lagging approximately 6 hours behind CYP7A1. We conclude that, in the rat, like CYP7A1, CYP7B1 demonstrates diurnal rhythm and is regulated by bile acids and cholesterol.
