Post-stroke treatment with argon attenuated brain injury, reduced brain inflammation and enhanced M2 microglia/macrophage polarization: a randomized controlled animal study

• Published in: Critical care (London, England) Vol. 23; no. 1; p. 198
• Main Authors: Liu, Jingjin, Nolte, Kay, Brook, Gary, Liebenstund, Lisa, Weinandy, Agnieszka, Höllig, Anke, Veldeman, Michael, Willuweit, Antje, Langen, Karl-Josef, Rossaint, Rolf, Coburn, Mark
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 03-06-2019; BioMed Central; BMC
• Subjects: Analysis of Variance; Animals; Apoptosis; Argon; Argon - pharmacology; Argon - therapeutic use; Brain; Brain damage; Brain injuries; Brain Injuries - etiology; Brain Injuries - pathology; Brain Injuries - physiopathology; Care and treatment; Cell culture; Consultants (Persons); Critical care; Cytokines; Disease Models, Animal; Encephalitis - physiopathology; Encephalitis - prevention & control; Gene expression; Genotype & phenotype; Health aspects; Immune system; Infarction, middle cerebral artery; Infarction, Middle Cerebral Artery - complications; Infarction, Middle Cerebral Artery - drug therapy; Infarction, Middle Cerebral Artery - physiopathology; Inflammation; Ischemia; Male; Microglia; Neurogenesis; Neurons; Neuroprotection; Neuroprotective Agents - pharmacology; Neuroprotective Agents - therapeutic use; Noble gases; Random Allocation; Rats; Rats, Wistar - injuries; Rodents; Stroke; Stroke - drug therapy; Stroke - pathology; Stroke - physiopathology; Surgery; Traumatic brain injury; Tumor necrosis factor-TNF; Germany; Noble gases; Neuroprotection; Infarction, middle cerebral artery; Argon
• ISSN: 1364-8535; 1466-609X; 1364-8535; 1366-609X
• DOI: 10.1186/s13054-019-2493-7

In recent years, argon has been shown to exert neuroprotective effects in an array of models. However, the mechanisms by which argon exerts its neuroprotective characteristics remain unclear. Accumulating evidence imply that argon may exert neuroprotective effects via modulating the activation and polarization of microglia/macrophages after ischemic stroke. In the present study, we analyzed the underlying neuroprotective effects of delayed argon application until 7 days after reperfusion and explored the potential mechanisms.
Twenty-one male Wistar rats underwent transient middle cerebral artery occlusion or sham surgery randomly for 2 h using the endoluminal thread model. Three hours after transient middle cerebral artery occlusion induction and 1 h after reperfusion, animals received either 50% vol Argon/50% vol O
or 50% vol N
/50% vol O
for 1 h. The primary outcome was the 6-point neuroscore from 24 h to d7 after reperfusion. Histological analyses including infarct volume, survival of neurons (NeuN) at the ischemic boundary zone, white matter integrity (Luxol Fast Blue), microglia/macrophage activation (Iba1), and polarization (Iba1/Arginase1 double staining) on d7 were conducted as well. Sample size calculation was performed using nQuery Advisor + nTerim 4.0. Independent t test, one-way ANOVA and repeated measures ANOVA were performed, respectively, for statistical analysis (SPSS 23.0).
The 6-point neuroscore from 24 h to d7 after reperfusion showed that tMCAO Ar group displayed significantly improved neurological performance compared to tMCAO N
group (p = 0.026). The relative numbers of NeuN-positive cells in the ROIs of tMCAO Ar group significantly increased compared to tMCAO N
group (p = 0.010 for cortex and p = 0.011 for subcortex). Argon significantly suppressed the microglia/macrophage activation as revealed by Iba1 staining (p = 0.0076) and promoted the M2 microglia/macrophage polarization as revealed by Iba1/Arginase 1 double staining (p = 0.000095).
Argon administration with a 3 h delay after stroke onset and 1 h after reperfusion significantly alleviated neurological deficit within the first week and preserved the neurons at the ischemic boundary zone 7 days after stroke. Moreover, argon reduced the excessive microglia/macrophage activation and promoted the switch of microglia/macrophage polarization towards the anti-inflammatory M2 phenotype. Studies making efforts to further elucidate the protective mechanisms and to benefit the translational application are of great value.