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. 2022 Oct 13;13(3):1128–1144. doi: 10.1016/j.apsb.2022.10.011

Figure 2.

Figure 2

GPR35 deficiency in hepatocytes exacerbates HFCF diet-induced steatohepatitis and insulin resistance (n = 10). C57BL/6J mice were injected (i.v.) with AAV8-TBG-SaCas9-2A-EGFP-U6-sgRNA (NC) or AAV8-TBG-SaCas9-2A-EGFP-U6-sgRNA (Gpr35) to generate control (Gpr35hep-NC1) mice and hepatocyte GPR35-knockout (Gpr35hep−/−) mice, which were then fed a NCD or HFCF 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 carried out on WT, Gpr35hep-NC1, and Gpr35hep−/− mice after NCD or HFCF diet feeding for 16 weeks. (D–N) WT, Gpr35hep-NC1, and Gpr35hep−/− 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, 20 μm. The mRNA expression of the genes was normalized to that of β-actin. Data are the mean ± SD; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.