Effects of ketogenic diet and ketone monoester supplement on acute alcohol withdrawal symptoms in male mice

Abstract

Rationale: After alcohol ingestion, the brain partly switches from consumption of glucose to consumption of the alcohol metabolite acetate. In heavy drinkers, the switch persists after abrupt abstinence, leading to the hypothesis that the resting brain may be "starved" when acetate levels suddenly drop during abstinence, despite normal blood glucose, contributing to withdrawal symptoms. We hypothesized that ketone bodies, like acetate, could act as alternative fuels in the brain and alleviate withdrawal symptoms.

Objectives: We previously reported that a ketogenic diet during alcohol exposure reduced acute withdrawal symptoms in rats. Here, our goals were to test whether (1) we could reproduce our findings, in mice and with longer alcohol exposure; (2) ketone bodies alone are sufficient to reduce withdrawal symptoms (clarifying mechanism); (3) introduction of ketogenic diets at abstinence (a clinically more practical implementation) would also be effective.

Methods: Male C57BL/6NTac mice had intermittent alcohol exposure for 3 weeks using liquid diet. Somatic alcohol withdrawal symptoms were measured as handling-induced convulsions; anxiety-like behavior was measured using the light-dark transition test. We tested a ketogenic diet, and a ketone monoester supplement with a regular carbohydrate-containing diet.

Results: The regular diet with ketone monoester was sufficient to reduce handling-induced convulsions and anxiety-like behaviors in early withdrawal. Only the ketone monoester reduced handling-induced convulsions when given during abstinence, consistent with faster elevation of blood ketones, relative to ketogenic diet.

Conclusions: These findings support the potential utility of therapeutic ketosis as an adjunctive treatment in early detoxification in alcohol-dependent patients seeking to become abstinent.

Trial registration: clinicaltrials.gov NCT03878225, NCT03255031.

Keywords: Alcohol dependence; Alcohol withdrawal; Alcoholism; Anxiety-like behavior; Detoxification; Ethanol; Ketone bodies; Ketone monoester; Mice.

Conflict of interest statement

The authors report no conflict of interest.

Figures

Fig. 1. Experimental setup and diets

(a) Timeline of alcohol exposure (grey) and forced abstinence (open), with test times as hours since alcohol removal shown for the last cycle. Periods with test diets are indicated by diagonal hatching. Arrows indicate blood alcohol sampling (pointing down) or light-dark transition test (pointing up). (b) Composition of diets as calories provided by each nutrient, per mL diet. Reg.: regular diet. Alco: alcohol-containing. All diets also contained vitamins, minerals, and 5.5–5.7% fiber.

Fig. 2. Blood ketone and glucose levels

Blood BHB as a function of time in the “Keto Throughout” experiment (a, n=12) and the “Keto After” experiment (b, n=10–12), blood glucose as a function of time in “Keto After” (c, n=10–12). *p

Fig. 3. Effects of diets on handling-induced convulsions

HIC score as AUC for the first three hours of abstinence (a) and total observation period (b) for no-alcohol controls, in “Keto Throughout” (n=12), and “Keto After” (n=10–12). **p

Fig. 4. Effects of diets in the light-dark transition test

Light-dark side crossings (a-c) and voluntary time in the lit side (d-f) in the “Keto Throughout” experiment (a,d, n=6), the “Keto After” experiment (b,e, n=10–11), and no-alcohol controls (c,f, n=6–12). *p

Fig. 5. Blood alcohol levels and control experiments

Blood alcohol in “Keto Throughout” and “Keto After” (a, n=5–6). Blood alcohol (b) and loss of righting reflex (c, LORR) after acute 3.5 g/kg alcohol administration, in mice fed no-alcohol regular, KD, or KME diet; n=6. Number of alcohol (20% in water, d) or liquid food (e) reinforcers taken per 2h-session at baseline and after intragastric administration of 3 g/kg KME or isocaloric oil control, n=7–9. Data are shown as individual subjects, with bars representing group means ± s.e.m.