A High-Cholesterol Diet Exacerbates Liver Fibrosis in Mice via Accumulation of Free Cholesterol in Hepatic Stellate Cells

Background & Aims

Some studies have indicated that dietary cholesterol has a role in the progression of liver fibrosis. We investigated the mechanisms by which dietary cholesterol might contribute to hepatic fibrogenesis.

Methods

C57BL/6 mice were fed a high-cholesterol diet or a control diet for 4 weeks; liver fibrosis then was induced by bile-duct ligation or carbon tetrachloride administration. Hepatic stellate cells (HSCs) were isolated from mice fed high-cholesterol diets or from Niemann–Pick type C1-deficient mice, which accumulate intracellular free cholesterol.

Results

After bile-duct ligation or carbon tetrachloride administration, mice fed high-cholesterol diets had significant increases in liver fibrosis and activation of HSCs compared with mice fed control diets. There were no significant differences in the degree of hepatocellular injury or liver inflammation, including hepatocyte apoptosis or Kupffer cell activation, between mice fed high-cholesterol or control diets. Levels of free cholesterol were much higher in HSCs from mice fed high-cholesterol diets than those fed control diets. In cultured HSCs, accumulation of free cholesterol in HSCs increased levels of Toll-like receptor 4 (TLR4), leading to down-regulation of bone morphogenetic protein and activin membrane-bound inhibitor (a pseudoreceptor for transforming growth factor [TGF]β); the HSCs became sensitized to TGFβ-induced activation. Liver fibrosis was not aggravated by the high-cholesterol diet in C3H/HeJ mice, which express a mutant form of TLR4; HSCs that express mutant TLR4 were not activated by accumulation of free cholesterol.

Conclusions

Dietary cholesterol aggravates liver fibrosis because free cholesterol accumulates in HSCs, leading to increased TLR4 signaling, down-regulation of bone morphogenetic protein and activin membrane-bound inhibitor, and sensitization of HSC to TGFβ. This pathway might be targeted by antifibrotic therapies.

Keywords:  Liver Disease , Mouse Model , Dyslipidemia , Lipopolysaccharide

Abbreviations used in this paper:  AcLDL, acetyl low-density lipoprotein, ALT, alanine aminotransferase, Bambi, BMP and activin membrane-bound inhibitor, BDL, bile duct ligation, CCl₄, carbon tetrachloride, CE, cholesterol ester, compound 58035, acyl-CoA:cholesterol acyltransferase inhibitor 58035, FC, free cholesterol, HC, high cholesterol, HSCs, hepatic stellate cells, KO, knock-out, LPS, lipopolysaccharide, mRNA, messenger RNA, NPC1, Niemann-Pick type C1, αSMA, α-smooth muscle actin, TC, total cholesterol, TG, triglyceride, TGFβ, transforming growth factor β, TLR4, Toll-like receptor 4, TNFα, tumor necrosis factor-α, WT, wild-type