25-Hydroxycholesterol activates the expression of cholesterol 25-hydroxylase in an LXR-dependent mechanism

Ying Liu,Zhuo Wei,Xingzhe Ma,Xiaoxiao Yang,Yuanli Chen,Lei Sun,Chuanrui Ma,Qing R Miao,David P Hajjar,Jihong Han,Yajun Duan

doi:10.1194/jlr.m080440

Abstract

Cholesterol 25-hydroxylase (CH25H) catalyzes the production of 25-hydroxycholesterol (25-HC), an oxysterol that can play an important role in different biological processes. However, the mechanisms regulating CH25H expression have not been fully elucidated. In this study, we determined that CH25H is highly expressed in mouse liver and peritoneal macrophages. We identified several liver X receptor (LXR) response elements (LXREs) in the human CH25H promoter. In HepG2 cells, activation of LXR by 25-HC or other oxysterols and synthetic ligands [T0901317 (T317) and GW3965] induced CH25H protein expression, which was associated with increased CH25H mRNA expression. 25-HC or T317 activated CH25H transcription in an LXRE-dependent manner. Thus, high-expressing LXRα or LXRβ activated CH25H expression, and the activation was further enhanced by LXR ligands. In contrast, inhibition of LXRα/β expression attenuated 25-HC or T317-induced CH25H expression. Deficiency of interferon γ expression reduced, but did not block, LXR ligand-induced hepatic CH25H expression. Activation of LXR also substantially induced macrophage CH25H expression. In vivo, administration of GW3965 to mice increased CH25H expression in both liver and peritoneal macrophages. Taken together, our study demonstrates that 25-HC can activate CH25H expression in an LXR-dependent manner, which may be an important mechanism to exert the biological actions of 25-HC.

Highlights

Cholesterol 25-hydroxylase (CH25H) catalyzes the production of 25-hydroxycholesterol (25-HC), an oxysterol that can play an important role in different biological processes
Several cytochrome P450 enzymes may complete the conversion of cholesterol into 25-HC in vitro, cholesterol 25-hydroxylase (CH25H) has been demonstrated to play a critical role in the production of 25-HC, both in vitro and in vivo [1]. 25-HC can play an important role in different biological processes, in cholesterol metabolism
Western blot analysis Expression of CH25H, liver X receptor (LXR), LXR, SREBP1, SREBP2, FASN, HMG-CoA reductase (HMGCR), ABCA1, ABCG1, and GAPDH protein was determined by Western blot with cellular proteins extracted from HepG2 cells, primary hepatocytes, RAW264.7 cells, peritoneal macrophages, or mouse liver as described [20]

Summary

Introduction

Cholesterol 25-hydroxylase (CH25H) catalyzes the production of 25-hydroxycholesterol (25-HC), an oxysterol that can play an important role in different biological processes. Western blot analysis Expression of CH25H, LXR , LXR , SREBP1, SREBP2, FASN, HMGCR, ABCA1, ABCG1, and GAPDH protein was determined by Western blot with cellular proteins extracted from HepG2 cells, primary hepatocytes, RAW264.7 cells, peritoneal macrophages, or mouse liver as described [20]. To determine whether LXR activation can induce CH25H expression in mouse liver and peritoneal macrophages, C57BL/6 mice (males, 8-week old) were randomly divided into two groups (5/group), and they received the following treatment for one week: control group mice were fed normal chow; GW3965 group mice were fed normal chow containing GW3965, a synthetic LXR ligand, at a dose of 20 mg/day/kg bodyweight.

Results

Conclusion