Acute Crit Care.  2020 Nov;35(4):286-295. 10.4266/acc.2020.00304.

Therapeutic hypothermia reduces inflammation and oxidative stress in the liver after asphyxial cardiac arrest in rats

• Park Y
• Ahn JH
• Lee TK
• Kim B
• Tae HJ
• Park JH
• Shin MC
• Cho JH
• Won MH

• KMID: 2510548
• DOI: http://doi.org/10.4266/acc.2020.00304

Abstract

Background
Few studies have evaluated the effects of hypothermia on cardiac arrest (CA)-induced liver damage. This study aimed to investigate the effects of hypothermic therapy on the liver in a rat model of asphyxial cardiac arrest (ACA).
Methods
Rats were subjected to 5-minute ACA followed by return of spontaneous circulation (RoSC). Body temperature was controlled at 33°C±0.5°C or 37°C±0.5°C for 4 hours after RoSC in the hypothermia group and normothermia group, respectively. Liver tissues in each group were collected at 6 hours, 12 hours, 1 day, and 2 days after RoSC. To examine hepatic inflammation, mast cells were stained with toluidine blue. Superoxide anion radical production was evaluated using dihydroethidium fluorescence straining and expression of endogenous antioxidants (superoxide dismutase 1 [SOD1] and SOD2) was examined using immunohistochemistry.
Results
There were significantly more mast cells in the livers of the normothermia group with ACA than in the hypothermia group with ACA. Gradual increase in superoxide anion radical production was found with time in the normothermia group with ACA, but production was significantly suppressed in the hypothermia group with ACA relative to the normothermia group with ACA. SOD1 and SOD2 levels were higher in the hypothermia group with ACA than in the normothermia group with ACA.
Conclusions
Experimental hypothermic treatment after ACA significantly inhibited inflammation and superoxide anion radical production in the rat liver, indicating that this treatment enhanced or maintained expression of antioxidants. Our findings suggest that hypothermic therapy after CA can reduce mast cell-mediated inflammation through regulation of oxidative stress and the expression of antioxidants in the liver.

Keyword

antioxidants; asphyxia; induced hypothermia; liver; oxidative stress

Figure

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  Figure 1. Toluidine blue staining of liver sections in the normothermia (NT; A-E) and hypithermia (HT; F-J) groups at sham, 6 hours, 12 hours, 1 day, and 2 days after asphyxial cardiac arrest (ACA). Few toluidine blue positive mast cells were observed in the NT/sham and HT/sham groups. Numbers of mast cells (arrows) were increased after ACA, with a peak in number 1 day post-ACA in the NT/ACA group. In the HT/ ACA group, the change in number of mast cells over time was similar to that observed in the NT/ACA group, but there were significantly fewer mast cells than in the corresponding NT/ACA group. CV: central vein. Scale bar=50 µm. (K) Mean numbers of toluidine blue positive mast cells (n=7 at each time; aP<0.05, significantly different from NT/sham group; bP<0.05, significantly different from the previous timepoint group; cP<0.05, significantly different from NT/ACA group). Bars indicate mean±standard error of the mean.

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  Figure 2. Dihydroethidium (DHE) fluorescence staining in liver sections of normothermia (NT; A-E) and hypithermia (HT; F-J) groups at sham, 6 hours, 12 hours, 1 day, and 2 days after asphyxial cardiac arrest (ACA). Very weak DHE fluorescence was observed in the NT/sham and HT/sham groups. DHE fluorescence in both NT/ACA and HT/ACA groups gradually increased over time, but DHE fluorescence in the HT/ ACA group was lower than that in the NT/ACA group. CV: central vein. Scale bar=50 µm. (K) Relative optical density (ROD) of DHE fluorescence in all groups (n=7 at each time after ACA, aP<0.05, significantly different from NT/sham group; bP<0.05; significantly different from previous time-point group; cP<0.05, significantly different from NT/ACA group). Bars indicate means±standard error of the mean.

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  Figure 3. Immunohistochemical staining for superoxide dismutase 1 (SOD1) in liver sections of normothermia (NT; A-E) and hypithermia (HT; F-J) groups at sham, 6 hours, 12 hours, 1 day, and 2 days after asphyxial cardiac arrest (ACA). SOD1 immunoreactivity decreased after 6 hours post-ACA in the NT/ACA group. SOD1 immunoreactivity was significantly higher in the HT/ACA group than in the NT/ACA group at all times after ACA. CV: central vein. Scale bar=50 µm. (K) Relative optical density (ROD) of SOD1 immunoreactivity in all groups (n=7 at each time after ACA, aP<0.05, significantly different from NT/sham group; bP<0.05, significantly different from previous time-point group; cP<0.05, significantly different from NT/ACA group). Bars indicate means±standard error of the mean.

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  Figure 4. Immunohistochemical staining for superoxide dismutase 2 (SOD2) in liver sections from the normothermia (NT; A-E) and hypithermia (HT; F-J) groups at sham, 6 hours, 12 hours, 1 day, and 2 days after asphyxial cardiac arrest (ACA). SOD2 immunoreactivity in the NT/ ACA group was markedly decreased 6 hours after ACA. In the HT/ACA group, SOD2 immunoreactivity did not change significantly after ACA. CV: central vein. Scale bar=50 µm. (F) Relative optical density (ROD) of SOD2 immunoreactivity in all groups (n=7 at each time after ACA, aP<0.05, significantly different from NT/sham group; bP<0.05, significantly different from previous time-point group; cP<0.05, significantly different from NT/ACA group). Bars indicate means±standard error of the mean.

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