Antioxidants that protect mitochondria reduce interleukin-6 and oxidative stress, improve mitochondrial function, and reduce biochemical markers of organ dysfunction in a rat model of acute sepsis

D A Lowes, N R Webster, M P Murphy, H F Galley, D A Lowes, N R Webster, M P Murphy, H F Galley

Abstract

Background: Sepsis-induced organ failure is the major cause of death in critical care units, and is characterized by a massive dysregulated inflammatory response and oxidative stress. We investigated the effects of treatment with antioxidants that protect mitochondria (MitoQ, MitoE, or melatonin) in a rat model of lipopolysaccharide (LPS) plus peptidoglycan (PepG)-induced acute sepsis, characterized by inflammation, mitochondrial dysfunction and early organ damage.

Methods: Anaesthetized and ventilated rats received an i.v. bolus of LPS and PepG followed by an i.v. infusion of MitoQ, MitoE, melatonin, or saline for 5 h. Organs and blood were then removed for determination of mitochondrial and organ function, oxidative stress, and key cytokines.

Results: MitoQ, MitoE, or melatonin had broadly similar protective effects with improved mitochondrial respiration (P<0.002), reduced oxidative stress (P<0.02), and decreased interleukin-6 levels (P=0.0001). Compared with control rats, antioxidant-treated rats had lower levels of biochemical markers of organ dysfunction, including plasma alanine amino-transferase activity (P=0.02) and creatinine concentrations (P<0.0001).

Conclusions: Antioxidants that act preferentially in mitochondria reduce mitochondrial damage and organ dysfunction and decrease inflammatory responses in a rat model of acute sepsis.

Figures

Fig 1
Fig 1
Schematic diagram of study design. Preparation=induction of anaesthesia with isoflurane in a tank, then a tracheostomy with isoflurane anaesthesia via a nose cone followed by ventilation, venous cannulation, and siting of ECG electrodes. Sampling=cardiac puncture, laparotomy, organ removal, termination.
Fig 2
Fig 2
(a) Plasma ALT activity, (b) plasma AST activity, and (c) plasma creatinine levels 5 h after administration of LPs/PepG or saline showing the effects of MitoE, MitoQ or melatonin (Mel). Box and whisker plots show the median, IQR, and full range. *Significantly higher than saline control; #significantly lower than LPS/PepG control (Mann–Whitney U-test). P-value shown is Kruskal–Wallis across the LPS/PepG groups.
Fig 3
Fig 3
(a) Liver protein carbonyl concentrations and (b) plasma lipid hydroperoxide (LPO) concentrations 5 h after administration of saline or LPS/PepG, and the effects of treatment with MitoE, MitoQ, or melatonin (Mel). Box and whisker plots show the median, IQR, and full range. *Significantly higher than saline control; #significantly lower than LPS/PepG control (Mann–Whitney U-test). P-value shown is Kruskal–Wallis across the LPS/PepG groups.
Fig 4
Fig 4
(a) Plasma IL-6 and (b) plasma IL-10 levels 5 h after administration of LPS/PepG or saline, and the effects of MitoE, MitoQ, or melatonin (Mel). Box and whisker plots show the median, IQR, and full range. *Significantly higher than saline control; #significantly lower and †significantly higher than LPS/PepG control (Mann–Whitney U-test). P-value shown is Kruskal–Wallis across the LPS/PepG groups.
Fig 5
Fig 5
Mitochondrial ATP:O ratios with (a) glutamate and malate as substrate (complexes I, II, III, and IV) and (b) succinate (complexes II, II, and IV) and (c) complex IV activity (as oxygen used per minute) 5 h after administration of saline or LPS/PepG and the effect of treatment with MitoE, MitoQ, or melatonin (Mel). Box and whisker plots show the median, IQR, and full range. *Significantly higher than saline control; #significantly higher than the LPS/PepG control (Mann–Whitney U-test). P-value shown is Kruskal–Wallis across LPS/PepG groups.

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