Identification of the toxic threshold of 3-hydroxybutyrate-sodium supplementation in septic mice

Background In septic mice, supplementing parenteral nutrition with 150 mg/day 3-hydroxybutyrate-sodium-salt (3HB-Na) has previously shown to prevent muscle weakness without obvious toxicity. The main objective of this study was to identify the toxic threshold of 3HB-Na supplementation in septic mice, prior to translation of this promising intervention to human use. Methods In a centrally-catheterized, antibiotic-treated, fluid-resuscitated, parenterally fed mouse model of prolonged sepsis, we compared with placebo the effects of stepwise escalating doses starting from 150 mg/day 3HB-Na on illness severity and mortality ( n = 103). For 5-day survivors, also the impact on ex-vivo -measured muscle force, blood electrolytes, and markers of vital organ inflammation/damage was documented. Results By doubling the reference dose of 150 mg/day to 300 mg/day 3HB-Na, illness severity scores doubled ( p = 0.004) and mortality increased from 30.4 to 87.5 % ( p = 0.002). De-escalating this dose to 225 mg still increased mortality ( p ≤ 0.03) and reducing the dose to 180 mg/day still increased illness severity ( p ≤ 0.04). Doses of 180 mg/day and higher caused more pronounced metabolic alkalosis and hypernatremia ( p ≤ 0.04) and increased markers of kidney damage ( p ≤ 0.05). Doses of 225 mg/day 3HB-Na and higher caused dehydration of brain and lungs ( p ≤ 0.05) and increased markers of hippocampal neuronal damage and inflammation ( p ≤ 0.02). Among survivors, 150 mg/day and 180 mg/day increased muscle force compared with placebo ( p ≤ 0.05) up to healthy control levels ( p ≥ 0.3). Conclusions This study indicates that 150 mg/day 3HB-Na supplementation prevented sepsis-induced muscle weakness in mice. However, this dose appeared maximally effective though close to the toxic threshold, possibly in part explained by excessive Na + intake with 3HB-Na. Although lower doses were not tested and thus might still hold therapeutic potential, the current results point towards a low toxic threshold for the clinical use of ketone salts in human critically ill patients. Whether 3HB-esters are equally effective and less toxic should be investigated.