Cardiac energetic impairment is a major finding in takotsubo patients. We investigate specific metabolic adaptations to direct future therapies.
Methods and Results
An isoprenaline-injection female rat model (versus sham) was studied at day-3; recovery assessed at day-7. Substrate uptake, metabolism, inflammation and remodelling were investigated by 18F-FDG-PET, metabolomics, qPCR and WB. Isolated cardiomyocytes were patch-clamped during stress protocols for redox states of NAD(P)H/FAD or [Ca2+]c, [Ca2+]m and sarcomere length. Mitochondrial respiration was assessed by seahorse/Clark electrode (glycolytic and β-oxidation substrates).
Cardiac 18F-FDG metabolic rate was increased in takotsubo (p = 0.006), as were expression of GLUT4-RNA/GLUT1/HK2-RNA and HK activity (all p < 0.05), with concomitant accumulation of glucose- and fructose-6-phosphates (p > 0.0001). Both lactate and pyruvate were lower (p < 0.05) despite increases in LDH-RNA and PDH (p < 0.05 both). β-oxidation enzymes CPT1b-RNA and 3KAT were increased (p < 0.01) but malonyl-CoA (CPT-1 regulator) was upregulated (p = 0.01) with decreased fatty acids and acyl-carnitines levels (p = 0.0001-0.02). Krebs cycle intermediates α-ketoglutarate and succinyl-carnitine were reduced (p < 0.05) as was cellular ATP reporter dihydroorotate (p = 0.003). Mitochondrial Ca2+ uptake during high workload was impaired on day-3 (p < 0.0001), inducing oxidation of NAD(P)H and FAD (p = 0.03) but resolved by day-7. There were no differences in mitochondrial respiratory function, sarcomere shortening or [Ca2+] transients of isolated cardiomyocytes, implying preserved integrity of both mitochondria and cardiomyocyte. Inflammation and remodelling were upregulated - increased CD68-RNA, collagen RNA/protein and skeletal actin RNA (all p < 0.05).
Dys-regulation of glucose and lipid metabolic pathways with decreases in final glycolytic and β-oxidation metabolites and reduced availability of Krebs intermediates characterises takotsubo myocardium. The energetic deficit accompanies defective Ca2+ handling, inflammation and upregulation of remodelling pathways, with preservation of sarcomeric and mitochondrial integrity.
The simultaneous dysregulation in the glycolytic and beta-oxidation pathways which underlies the energetic deficit of the takotsubo heart supports further testing of currently available metabolic modulators as possible candidates for successful therapy, as well as targeting the inflammatory and remodelling pathways.