Methane Saline Ameliorates Traumatic Brain Injury through Anti-Inflammatory, Antiapoptotic, and Antioxidative Effects by Activating the Wnt Signalling Pathway

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Methane Saline Ameliorates Traumatic Brain Injury through Anti-Inflammatory, Antiapoptotic, and Antioxidative Effects by Activating the Wnt Signalling Pathway

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Meng Li,¹ Weiman Gao,¹ Le Ji,² Jia Li,¹ Wanting Jiang,³ and Wenchen Ji¹

¹Department of Orthopedics, The First Affiliated Hospital of Xi’an Jiaotong University, 710061, China

²Department of Orthopedics, Shaanxi Provincial People's Hospital, 710068, China

³Department of Ultrasound Diagnosis, Xi’an People’s Hospital (The Fourth Hospital of Xi’an), 710004, China

Academic Editor: Anna Chiarini

Received02 Apr 2020

Revised25 Aug 2020

Accepted13 Nov 2020

Published18 Dec 2020

Abstract

Objective. 

Methane saline (MS) can be used to treat many diseases via its anti-inflammatory, antiapoptotic, and antioxidative activities. However, to date, there is no published evidence as to whether MS has any effect on traumatic brain injury (TBI). The Wnt signalling pathway regulates cell proliferation, differentiation, migration, and apoptosis; however, whether the Wnt signalling pathway regulates any effect of MS on TBI is unknown. This study was designed to explore the role of MS in the treatment of TBI and whether the Wnt pathway is involved.  

Methods. 

Sprague-Dawley rats were randomly divided into five groups: sham, TBI, TBI+10 ml/kg MS, TBI+20 ml/kg MS, and TBI+30 ml/kg MS. After induction of TBI, MS was injected intraperitoneally once daily for seven consecutive days. Neurological function was evaluated by the Neurological Severity Score (NSS) at 1, 7, and 14 days after TBI. Haematoxylin-eosin (HE) staining, inflammatory factors, neuron-specific enolase (NSE) staining, oxidative stress, and cell apoptosis were measured and compared 14 d after TBI to identify the optimal dose of MS and to investigate the effect of MS on TBI. In the second experiment, Sprague-Dawley rats were randomly divided into four groups: sham, TBI, TBI+20 ml/kg MS, and TBI+20 ml/kg MS+Dickkopf-1 (DKK-1, a specific inhibitor of the Wnt pathway). NSE, caspase-3, superoxide dismutase (SOD), Wnt3a, and β-catenin were detected by real-time PCR and Western blotting. The results from each group were compared 14 d after TBI to determine the regulatory role of the Wnt pathway.  

Results. 

Methane saline significantly inhibited inflammation, oxidative stress, and cell apoptosis, thus protecting neurons within 14 days of TBI. The best treatment effect against TBI was obtained with 20 ml/kg MS. When the Wnt pathway was inhibited, the treatment effect of MS was impaired.  

Conclusion. Methane saline ameliorates TBI through its anti-inflammatory, antiapoptotic, and antioxidative effects via activation of the Wnt signalling pathway, which plays a part but is not the only mechanism underlying the effects of MS. Thus, MS may be a novel strategy for treating TBI.

1. Introductions

Traumatic brain injury (TBI) is a common disease that is primarily caused by car accidents and high-altitude falls. It can result in neurological dysfunction and death. TBI has two clinical phases: primary and secondary injury [1]. The former involves deformation of the brain caused by a direct force; once this happens, it is irreversible [2]. The latter phase is the cascade of responses that occur several hours after the primary injury, among which inflammatory responses and oxidative stress are the main factors underlying the local microenvironmental disorder [3]. Alleviation of these responses is essential for a functional recovery.

Methane has been extensively studied ever since its discovery in 1778 [4]. It was previously thought that highly concentrated methane (methane/air volume>30%) caused poisoning while low concentrations have no physiological effects [5, 6]. However, recently, it has been shown that methane has biological properties, especially anti-inflammation, antioxidative, and antiapoptotic effects and can be used to treat acute lung injury, autoimmune hepatitis, and retinal ischaemia injury [7–9]. The treatment mechanisms appear to be related to the regulation of mitochondrial function and proteins involved in signalling pathways [10, 11]. Given the volatility of methane, methane saline (MS), which is prepared under high-pressure conditions and has similar properties to methane, is used for treatment and research due to its ease of transport. However, it is not known whether MS can treat TBI and what signalling pathways are involved in the potential effects of MS on TBI. The Wnt pathway plays an important antiapoptotic role, maintains neuron survival, and modulates antioxidative stress [12]. Here, we determined whether MS has a therapeutic effect on TBI and the role of the Wnt pathway. This is the first time that MS has been used to treat TBI and that the regulatory mechanism has been described, thus building a theoretical basis for clinical use of MS.