Figure 3.

Mice with overexpressed GPR35 in hepatocytes are resistant to HFCF diet-induced steatohepatitis and IR (n = 10). C57BL/6J mice were injected (i.v.) with AAV8-TBGp-MCS-EGFP-3Flag-SV40 PolyA or AAV8-TBG-MCS-Gpr35-EGFP-3Flag-SV40 PolyA to generate control (Gpr35^hep-NC2) mice and hepatocyte GPR35-overexpressing (Gpr35^hep-OE) mice, which were then fed a NCD or HFCF diet for 16 weeks. (A) Body weight was measured every 4 weeks from 0 to 16 weeks. (B) Fasting blood glucose levels were measured every 4 weeks from 0 to 16 weeks. (C) GTTs and ITTs were undertaken on WT, Gpr35^hep-NC2, and Gpr35^hep-OE mice after NCD or HFCF-diet feeding for 16 weeks. (D–N) WT, Gpr35^hep-NC2, and Gpr35^hep-OE mice at the end of 16 weeks of HFCF-diet feeding. (D) Serum TG levels. (E) Liver weight. (F) TG and (G) FFA levels in the liver. (H) H&E staining of liver tissue sections. Scale bar, 50 μm. (I) Oil Red O staining of liver tissue sections. Scale bar, 20 μm. (J) Relative mRNA expression of Pparα and Cpt1α in the liver. (K) Serum levels of ALT and AST. (L) Relative mRNA expression of cytokines (Tnfα, Il6, Il1β) in the liver. (M) Representative images of F4/80 and MPO immunostaining of liver tissue sections. Scale bar, 20 μm. (N) Representative images of Sirius Red staining and α-SMA immunostaining of liver tissue sections. Scale bar, 50 μm. mRNA expression of genes was normalized to that of β-actin. The data are the mean ± SD. ns, not significant; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.