Prenatal Choline Supplementation Improves Glucose Tolerance and Reduces Liver Fat Accumulation in Mouse Offspring Exposed to Ethanol during the Prenatal and Postnatal Periods

Abstract

Prenatal alcohol exposure (AE) affects cognitive development. However, it is unclear whether prenatal AE influences the metabolic health of offspring and whether postnatal AE exacerbates metabolic deterioration resulting from prenatal AE. Choline is a semi-essential nutrient that has been demonstrated to mitigate the cognitive impairment of prenatal AE. This study investigated how maternal choline supplementation (CS) may modify the metabolic health of offspring with prenatal and postnatal AE (AE/AE). C57BL/6J female mice were fed either a Lieber-DeCarli diet with 1.4% ethanol between embryonic day (E) 9.5 and E17.5 or a control diet. Choline was supplemented with 4 × concentrations versus the control throughout pregnancy. At postnatal week 7, offspring mice were exposed to 1.4% ethanol for females and 3.9% ethanol for males for 4 weeks. AE/AE increased hepatic triglyceride accumulation in male offspring only, which was normalized by prenatal CS. Prenatal CS also improved glucose tolerance compared to AE/AE animals. AE/AE suppressed hepatic gene expression of peroxisome proliferator activated receptor alpha (Ppara) and low-density lipoprotein receptor (Ldlr), which regulate fatty acid catabolism and cholesterol reuptake, respectively, in male offspring. However, these changes were not rectified by prenatal CS. In conclusion, AE/AE led to an increased risk of steatosis and was partially prevented by prenatal CS in male mice.

Keywords: choline; glucose tolerance; prenatal alcohol exposure; steatosis.

Figure 1

Study design. C57BL/6J female mice received either a control or choline-supplemented diet with or without ethanol during pregnancy. Their offspring received either a control or ethanol-containing Lieber–DeCarli liquid diet after weaning for 4 weeks. AE: alcohol exposure; CS: choline supplementation; Ctrl: control. F: female, M: males; n = 6–8 offspring/sex/treatment group from 5 dams per group.

Figure 2

Food intake, weight gain, and gonadal fat weight of offspring mice exposed to ethanol with or without prenatal choline supplementation. C57BL/6J female mice received either a control or choline-supplemented diet with or without 1.4% ethanol during pregnancy. Their offspring (shown in this figure) received either a control or ethanol-containing (1.4% for females and 3.9% for males) Lieber–DeCarli liquid diet after weaning for 4 weeks. (A,B) average food intake of offspring during the 4-week post-weaning liquid diet feeding; (C,D) weight gain of offspring during the 4-week post-weaning liquid diet feeding; and (E,F) gonad fat weight of offspring after the 4-week post-weaning liquid diet feeding. Analyzed with one-way ANOVA. * p < 0.05. n = 6–8 offspring/sex/treatment group. AE: alcohol exposure; CS: choline supplementation; Ctrl: control. Ctrl/Ctrl: absolute control without alcohol exposure; Ctrl/AE: prenatal control and postnatal alcohol exposure; AE/AE: prenatal and postnatal alcohol exposure; AE-CS/AE: prenatal and postnatal alcohol exposure with prenatal choline supplementation.

Figure 3

Glucose tolerance of offspring mice exposed to ethanol with or without prenatal choline supplementation. C57BL/6J female mice received either a control or choline-supplemented diet with or without 1.4% ethanol during pregnancy. Their offspring (shown in this figure) received either a control or ethanol-containing (1.4% for females and 3.9% for males) Lieber–DeCarli liquid diet after weaning for 4 weeks. Intraperitoneal glucose tolerance test (GTT) was conducted by intraperitoneal injection of 2 mg/g body weight of glucose and blood glucose monitoring in the following 2 h. (A,B) male offspring GTT curve and area under the curve (AUC) after 4-week post-weaning liquid diet feeding; (C,D) female offspring GTT curve and area under the curve after 4-week post-weaning liquid diet feeding. Analyzed with repeated measures ANOVA. * p < 0.05 AE-CS/AE compared to AE/AE for males and AE-CS/AE compared to Ctrl/AE for females. n = 6–8 offspring/sex/treatment group. AE: alcohol exposure; CS: choline supplementation; Ctrl: control. Ctrl/Ctrl: absolute control without alcohol exposure; Ctrl/AE: prenatal control and postnatal alcohol exposure; AE/AE: prenatal and postnatal alcohol exposure; AE-CS/AE: prenatal and postnatal alcohol exposure with prenatal choline supplementation.

Figure 4

Liver gene expression of offspring mice exposed to ethanol with or without prenatal choline supplementation. C57BL/6J female mice received either a control or choline-supplemented diet with or without 1.4% ethanol during pregnancy. Their offspring (shown in this figure) received either a control or ethanol-containing (1.4% for females and 3.9% for males) Lieber–DeCarli liquid diet after weaning for 4 weeks. (A,C) male offspring liver gene expression after 4-week post-weaning liquid diet feeding; (B,D) female offspring liver gene expression after 4-week post-weaning liquid diet feeding. Analyzed with one-way ANOVA. * p < 0.05 compared to the Ctrl/Ctrl group. n = 6–8 offspring/sex/treatment group. AE: alcohol exposure; CS: choline supplementation; Ctrl: control. Ctrl/Ctrl: absolute control without alcohol exposure; Ctrl/AE: prenatal control and postnatal alcohol exposure; AE/AE: prenatal and postnatal alcohol exposure; AE-CS/AE: prenatal and postnatal alcohol exposure with prenatal choline supplementation. Acox1, Acyl-CoA oxidase 1; Bhmt1, betaine-homocysteine S-methyltransferase; Cd36, cluster of differentiation 36; Cpt1a, carnitine palmitoyltransferase 1a; Fasn, fatty acid synthase; Fatp1, fatty acid transporter 1; Ldlr, low-density lipoprotein receptor; Lpl, lipoprotein lipase; Pcyt1a, choline-phosphate cytidylyltransferase; Pemt, phosphatidylethanolamine N-methyltransferase; Ppara, peroxisome proliferator-activated receptor alpha.