Figure 2. Small molecule choline trimethylamine (TMA) lyase inhibition with iodomethylcholine (IMC) protects mice against ethanol-induced liver injury.

Nine- to eleven-week-old female C57BL6/J mice were fed either ethanol-fed or pair-fed in the presence and absence of IMC as described in the methods. Plasma levels of TMA (A), trimethylamine N-oxide (TMAO) (B), choline (C), carnitine (D), and betaine (E) were measured by mass spectrometry (n = 4–5). Plasma alanine aminotransferase (ALT) (F) was measured enzymatically (n = 4–5). Liver triglycerides (G), total cholesterol (H), cholesterol esters (I), and free cholesterol (J) were measured enzymatically (n = 4–5). (K) Representative H&E staining of livers from pair and EtOH-fed mice in the presence and absence of IMC. (L) Hepatic messenger RNA levels of tumor necrosis factor alpha (Tnfα). Statistics were completed by a two-way analysis of variance (ANOVA) followed by a Tukey’s multiple comparison test. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001. All data are presented as mean ± SEM, unless otherwise noted.

Figure 2—figure supplement 1. Small molecule inhibition with iodomethylcholine (IMC), but not fluoromethylcholine (FMC), reduces food intake in ethanol-fed mice.

Panels A–B and C–D represent data from IMC- and FMC-treated pair and ethanol-fed mice, respectively. (A, C) Body weights were measured biweekly throughout the 24-day experiment. #p ≤ 0.05 comparing pair to pair + IMC; ****p ≤ 0.0001 comparing ethanol to ethanol + IMC. (B, D) Diet intake was recorded daily throughout the experiment (n = 3 cages of six mice per group). The ethanol percentages (1–6%) are listed on top of the figures. *p ≤ 0.05 for days 6–22; n = 4–6. Statistics were completed using a Student’s t-test compared to the control mice. All data are presented as mean ± SEM, unless otherwise noted.