Research Article: Differential Feedback Regulation of Δ4-3-Oxosteroid 5β-Reductase Expression by Bile Acids

Date Published: January 26, 2017

Publisher: Public Library of Science

Author(s): Leila Valanejad, Christina Nadolny, Stephanie Shiffka, Yuan Chen, Sangmin You, Ruitang Deng, Gianfranco D. Alpini.


Δ4-3-oxosteroid 5β-reductase is member D1 of the aldo-keto reductase family 1 (AKR1D1), which catalyzes 5β-reduction of molecules with a 3-oxo-4-ene structure. Bile acid intermediates and most of the steroid hormones carry the 3-oxo-4-ene structure. Therefore, AKR1D1 plays critical roles in both bile acid synthesis and steroid hormone metabolism. Currently our understanding on transcriptional regulation of AKR1D1 under physiological and pathological conditions is very limited. In this study, we investigated the regulatory effects of primary bile acids, chenodeoxycholic acid (CDCA) and cholic acid (CA), on AKR1D1 expression. The expression levels of AKR1D1 mRNA and protein in vitro and in vivo following bile acid treatments were determined by real-time PCR and Western blotting. We found that CDCA markedly repressed AKR1D1 expression in vitro in human hepatoma HepG2 cells and in vivo in mice. On the contrary, CA significantly upregulated AKR1D1 expression in HepG2 cells and in mice. Further mechanistic investigations revealed that the farnesoid x receptor (FXR) signaling pathway was not involved in regulating AKR1D1 by bile acids. Instead, CDCA and CA regulated AKR1D1 through the mitogen-activated protein kinases/c-Jun N-terminal kinases (MAPK/JNK) signaling pathway. Inhibition of the MAPK/JNK pathway effectively abolished CDCA and CA-mediated regulation of AKR1D1. It was thus determined that AKR1D1 expression was regulated by CDCA and CA through modulating the MAPK/JNK signaling pathway. In conclusion, AKR1D1 expression was differentially regulated by primary bile acids through negative and positive feedback mechanisms. The findings indicated that both bile acid concentrations and compositions play important roles in regulating AKR1D1 expression, and consequently bile acid synthesis and steroid hormone metabolism.

Partial Text

Bile acids are the ultimate metabolites of cholesterol. Studies in the past decade uncovered a broad spectrum of functions associated with bile acids as hormone-like signaling molecules through various nuclear receptors, notably farnesoid x receptor (FXR) and G protein-coupled bile acid receptor 5 (TGR5) [1–3]. Both FXR and TGR5 signaling pathways play critical roles in regulating bile acids, cholesterol, lipids and glucose homeostasis [4–7]. Bile acid homeostasis is maintained through tightly regulated bile acid synthesis and enterohepatic circulation. Cholesterol 7α-hydroxylase (CYP7A1) is the rate limiting enzyme in classical bile acid synthesis while sterol 12α-hydroxylase (CYP8B1) is the determinant enzyme for the production of CA [8–11]. Canalicular secretion of bile acids through bile salt export pump (BSEP) is the rate-limiting step in the enterohepatic circulation of bile acids [12, 13]. Activation of FXR by bile acids represses CYP7A1 while induces BSEP expression to maintain hepatic bile acid homeostasis [14, 15].

In the bile acid synthesis pathway, AKR1D1 catalyzes the 5β-reduction of bile acid intermediates to eventually produce the primary bile acids CDCA and CA. It is well established that CYP7A1-mediated 7α-hydroxylation is the rate-limiting step in bile acid synthesis and bile acid homeostasis is maintained by regulating CYP7A1 expression through several negative feedback mechanisms [8–10]. In this study, we found that AKR1D1 was differentially regulated by the two primary bile acids. Similar to the effects on CYP7A1, CDCA markedly repressed AKR1D1 expression (Figs 1 and 3). On the other hand, CA induced AKR1D1 expression while CYP7A1 expression was minimally affected by CA (Figs 4 and 6). Our new findings indicate that bile acid synthesis is regulated at multiple steps in the bile acid synthesis pathway. In addition to CYP7A1, AKR1D1-mediated 5β-reduction of bile acid intermediates is also regulated by bile acids. CDCA-mediated repression of AKR1D1 expression represents a negative feedback mechanism to restrain bile acid production while CA-mediated upregulation of AKR1D1 represents a feed-forward regulation loop to promote bile acid production (Fig 12). The data thus suggest that both bile acid concentration and composition have an impact on controlling bile acid synthesis.