Figure 6.

Liver ILC1 augment alcohol-induced steatohepatitis via IL17 production. (A) Heat map shows mRNA expression of cytokines in liver ILC1 and NK cells from RNA-seq data GSE43339. Mice were treated as described in Figure 4F. (B and C) The mRNA level of IL13 (B) and IL17A (C) in liver tissue was detected (n = 4/group). (D) Plasma concentration of IL17A was measured (n = 4–7/group). (E) Expression of IL17A in liver ILC1 from CD- or EtOH-fed mice was measured by intracellular staining (n = 6/group). (F and G) The number of hepatic IL17A⁺ NK cells (F) and IL17A⁺ CD4⁺ T cells (G) in CD- or EtOH-fed mice were shown (n = 4/group). (H–J) EtOH-fed mice were injected intraperitoneally with IgG as control or anti-IL17A antibody twice at days 5 and 2 before binge (400 μg per mouse, n = 4/group). Liver tissue and plasma were obtained at 6 hours after binge. Hepatic triglyceride content (H), plasma level of ALT and AST (I), and H&E or Oil Red O staining of liver section (J) were examined. Representative images are shown (scale bars, 100 μm). (K) AML-12 cell line was stimulated with PBS or free fatty acid (FFA) (1 mmol/L) with or without IL17A (40 ng/mL). After 24 hours, the mRNA level of Srebp-1c and Acc in AML-12 cells was measured by quantitative polymerase chain reaction. Data were analyzed according to 3 independent experiments. One-way analysis of variance was performed in B–D. Two-tailed unpaired Student t tests were performed in E–I. Two-way analysis of variance was performed in K. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ∗∗∗∗P <.0001.