Abstract
The nuclear receptors farnesoid X receptor (FXR) and peroxisome proliferator-activated receptor (PPAR)γ exert counter-regulatory effects on hepatic stellate cells (HSCs) and protect against liver fibrosis development in rodents. Here, we investigated whether FXR ligands regulate PPARγ expression in HSCs and models of liver fibrosis induced in rats by porcine serum and carbon tetrachloride administration and bile duct ligation. Our results demonstrate that HSCs trans-differentiation associated with suppression of PPARγ mRNA expression, whereas FXR mRNA was unchanged. Exposure of cells to natural and synthetic ligands of FXR, including 6-ethyl chenodeoxycholic acid (6-ECDCA), a synthetic derivative of chenodeoxycholic acid, reversed this effect and increased PPARγ mRNA by≈ 40-fold. Submaximally effective concentrations of FXR and PPARγ ligands were additive in inhibiting α1(I) collagen mRNA accumulation induced by transforming growth factor (TGF)β1. Administration of 6-ECDCA in rats rendered cirrhotic by porcine serum and carbon tetrachloride administration or bile duct ligation reverted down-regulation of PPARγ mRNA expression in HSCs. Cotreatment with 6-ECDCA potentiates the antifibrotic activity of rosiglitazone, a PPARγ ligand, in the porcine serum model as measured by morphometric analysis of liver collagen content, hydroxyproline, and liver expression of α1(I) collagen mRNA,α -smooth muscle actin, fibronectin, TGFβ1, and tissue inhibitor of metalloprotease 1 and 2, whereas it enhanced the expression of PPARγ and uncoupling protein 2, a PPARγ-regulated gene, by 2-fold. In conclusion, by using an in vitro and in vivo approach, we demonstrated that FXR ligands up-regulate PPARγ mRNA in HSCs and in rodent models of liver fibrosis. A FXR-PPARγ cascade exerts counter-regulatory effects in HSCs activation.
Footnotes
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This study was partially supported by a research grant from Intercept Pharmaceuticals (New York, NY).
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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doi:10.1124/jpet.105.085597.
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ABBREVIATIONS: HSC, hepatic stellate cell; α-SMA,α -smooth muscle actin; NR, nuclear receptor; FXR, farnesoid X receptor; SHP, small heterodimer partner; TGF, transforming growth factor; PPAR, proliferator-activated receptor; 6-ECDCA, 6-ethyl chenodoexyxholic acid; GW4064, 3-(2,6-dichlorophenyl)-4-(3′-carboxy-2-chloro-stilben-4-yl)-oxymethyl-5-isopropyl-isoxazole; GW9662, 2-chloro-5-nitrobenbanilide; CDCA, chenodeoxycholic acid; TIMP, tissue inhibitor of matrix metalloproteinase; MMP, matrix metalloproteinase; qRT-PCR, quantitative reverse transcription-polymerase chain reaction; CMC, carboxymethyl cellulose; BDL, bile duct ligation; Wy-14,643, (4-chloro-6-[(2,3-dimethylphenyl)amino]-2-pyrimidinyl)thioacetic acid (pirinixic acid).
- Received March 15, 2005.
- Accepted June 20, 2005.
- The American Society for Pharmacology and Experimental Therapeutics
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