int-777 and Liver-Diseases

Hepatic ischemia/reperfusion injury (IRI) still remains an unavoidable problem in hepatectomy. The inflammatory response plays an important role in its pathogenesis. The plasma membrane-bound G protein-coupled bile acid receptor (TGR5), as one of G protein-coupled receptor (GPCR) families, has been proved to serve a protective role in several liver diseases. However, the exact function of TGR5 in modulating IRI remains obscure. We injected wild mice with a small interfering RNA of TGR5 (si-TGR5) or TGR5 agonist (INT-777) and established liver partial warm ischemia/reperfusion model. The results showed that knockdown of TGR5 significantly aggravated hepatic tissue injury, but treatment with INT-777 could reverse it, as evidenced by serum ALT and AST tests, liver histological injury, cytokines expressions, liver immunohistochemical analysis, and TUNEL staining. The apoptosis-associated proteins were evaluated after reperfusion. Moreover, we used primary bone marrow-derived macrophages (BMDMs) to establish hypoxia/reoxygenation (H/R) model to verify the anti-inflammation effect of TGR5. In in vivo experiments, we used TGR5-siRNA and TGR5 agonist (INT-777) to determine that TGR5 significantly attenuated liver damage after IRI through activating the Keap1-Nrf2 pathway. In addition, we found that overexpression of INT-777-activated TGR5 could reduce oxidative stress and inflammatory response in H/R-induced BMDMs through regulation of Keap1-Nef2 pathway during in vitro experiment. Importantly, these results were completely reversed in si-TGR5 BMDMs. In conclusion, the results indicated that TGR5 could effectively alleviated inflammation response via accelerating the activation of Keap1-Nrf2 signaling pathway during hepatic IRI, which may be meaningful in reducing related inflammatory molecules and adjusting inherent immunity.

Topics: Animals; Apoptosis; Cell Hypoxia; Cells, Cultured; Cholic Acids; Cytokines; Disease Models, Animal; Kelch-Like ECH-Associated Protein 1; Liver; Liver Diseases; Macrophages; Male; Mice, Inbred C57BL; NF-E2-Related Factor 2; Oxidative Stress; Receptors, G-Protein-Coupled; Reperfusion Injury; RNA Interference; Signal Transduction; Warm Ischemia

Chronic cholangiopathies have limited therapeutic options and represent an important indication for liver transplantation. The nuclear farnesoid X receptor (FXR) and the membrane G protein-coupled receptor, TGR5, regulate bile acid (BA) homeostasis and inflammation. Therefore, we hypothesized that activation of FXR and/or TGR5 could ameliorate liver injury in Mdr2(-/-) (Abcb4(-/-)) mice, a model of chronic cholangiopathy. Hepatic inflammation, fibrosis, as well as bile secretion and key genes of BA homeostasis were addressed in Mdr2(-/-) mice fed either a chow diet or a diet supplemented with the FXR agonist, INT-747, the TGR5 agonist, INT-777, or the dual FXR/TGR5 agonist, INT-767 (0.03% w/w). Only the dual FXR/TGR5 agonist, INT-767, significantly improved serum liver enzymes, hepatic inflammation, and biliary fibrosis in Mdr2(-/-) mice, whereas INT-747 and INT-777 had no hepatoprotective effects. In line with this, INT-767 significantly induced bile flow and biliary HCO 3- output, as well as gene expression of carbonic anhydrase 14, an important enzyme able to enhance HCO 3- transport, in an Fxr-dependent manner. In addition, INT-767 dramatically reduced bile acid synthesis via the induction of ileal Fgf15 and hepatic Shp gene expression, thus resulting in significantly reduced biliary bile acid output in Mdr2(-/-) mice.. This study shows that FXR activation improves liver injury in a mouse model of chronic cholangiopathy by reduction of biliary BA output and promotion of HCO 3--rich bile secretion.

Topics: Adenosine Triphosphatases; Analysis of Variance; Animals; Anion Transport Proteins; ATP Binding Cassette Transporter, Subfamily B; ATP-Binding Cassette Sub-Family B Member 4; Bile Acids and Salts; Biliary Tract Diseases; Cholic Acids; Disease Models, Animal; Liver Diseases; Male; Mice; Mice, Inbred C57BL; Random Allocation; Receptors, Cytoplasmic and Nuclear; Receptors, G-Protein-Coupled; Statistics, Nonparametric