Mitochondrial function in skeletal muscle of patients with protracted critical illness and ICU-acquired weakness

Kateřina Jiroutková, Adéla Krajčová, Jakub Ziak, Michal Fric, Petr Waldauf, Valér Džupa, Jan Gojda, Vlasta Němcova-Fürstová, Jan Kovář, Moustafa Elkalaf, Jan Trnka, František Duška, Kateřina Jiroutková, Adéla Krajčová, Jakub Ziak, Michal Fric, Petr Waldauf, Valér Džupa, Jan Gojda, Vlasta Němcova-Fürstová, Jan Kovář, Moustafa Elkalaf, Jan Trnka, František Duška

Abstract

Background: Mitochondrial damage occurs in the acute phase of critical illness, followed by activation of mitochondrial biogenesis in survivors. It has been hypothesized that bioenergetics failure of skeletal muscle may contribute to the development of ICU-acquired weakness. The aim of the present study was to determine whether mitochondrial dysfunction persists until protracted phase of critical illness.

Methods: In this single-centre controlled-cohort ex vivo proof-of-concept pilot study, we obtained vastus lateralis biopsies from ventilated patients with ICU-acquired weakness (n = 8) and from age and sex-matched metabolically healthy controls (n = 8). Mitochondrial functional indices were measured in cytosolic context by high-resolution respirometry in tissue homogenates, activities of respiratory complexes by spectrophotometry and individual functional capacities were correlated with concentrations of electron transport chain key subunits from respiratory complexes II, III, IV and V measured by western blot.

Results: The ability of aerobic ATP synthesis (OXPHOS) was reduced to ~54% in ICU patients (p<0.01), in correlation with the depletion of complexes III (~38% of control, p = 0.02) and IV (~26% of controls, p<0.01) and without signs of mitochondrial uncoupling. When mitochondrial functional indices were adjusted to citrate synthase activity, OXPHOS and the activity of complexes I and IV were not different, whilst the activities of complexes II and III were increased in ICU patients 3-fold (p<0.01) respectively 2-fold (p<0.01).

Conclusions: Compared to healthy controls, in ICU patients we have demonstrated a ~50% reduction of the ability of skeletal muscle to synthetize ATP in mitochondria. We found a depletion of complex III and IV concentrations and relative increases in functional capacities of complex II and glycerol-3-phosphate dehydrogenase/complex III.

Figures

Fig. 1
Fig. 1
An example of high-resolution respirometry assay in a homogenate of skeletal muscle, Protocol 1. Solid line represents oxygen consumption rate, dashed line oxygen concentration. Mal/Glu malate/glutamate, suc succinate, oligo oligomycin, FCCP uncoupler, AA antimycin A
Fig. 2
Fig. 2
Concentrations of functional subunits of respiratory complexes in arbitrary units and an example of an immunoblot membrane. Data are presented as medians, vertical bars represent interquartile ranges. GAPDH glyceraldehyde 3-phosphate dehydrogenase, COX cytochrome c oxidase
Fig. 3
Fig. 3
Activity of individual respiratory complexes adjusted to mitochondrial content (citrate synthase activity) measured by two independent methods. Upper row complex activity in cytosolic context determined by high-resolution respirometry in skeletal muscle homogenates. Lower row spectrophotometric analysis of the activity of individual respiratory complexes. Lines represent medians

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