Fig. 7
Pharmacological GPR65 inhibition alleviates hepatic fibrosis induced by CCl4. a BALB/c mice were divided into 4 groups randomly: Ctrl, Ctrl + CCl4, GPR65 inhibitor and GPR65 inhibitor + CCl4. The GPR65 inhibitor (10 mg/kg) was administered 4 weeks after the first CCl4 injection. Mice were administered CCl4 for 8 weeks. Hepatic fibrosis was evaluated by HE staining, Sirius red staining, Masson’s trichrome staining and IHC for COL1α1 and α-SMA. Scale bar = 100 μm for 40 × and 400 μm for 10 × . b qRT-PCR was used to assess the mRNA level of Gpr65, Acta2, Col1α1, Col3α1, Col12α1, Mmp2, Tgfβ1 and Pdgfβ (n = 3, 5, 3, 5). c Western blotting was used to determine the protein level of COL1α1, α-SMA, vimentin, MMP2 and TIMP1. d The content of hepatic hydroxyproline was quantified in livers of each group (n = 5, 7, 5, 7). Serum ALT (e) and AST (f) were examined (n = 5, 7, 5, 7). *P < 0.05 vs. Ctrl; #P < 0.05 for vs. Ctrl + CCl4. CCl4 carbon tetrachloride, Ctrl control, IHC immunohistochemistry, qRT-PCR quantitative real-time reverse transcription-polymerase chain reaction, COL1α1 collagen type I alpha 1, α-SMA α-smooth muscle actin, MMP2 matrix metalloproteinase 2, TIMP1 tissue inhibitor of metalloproteinases 1, ALT alanine aminotransferase, AST aspartate aminotransferase