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

WHOM is your doctors and stroke hospital contacting to get this tested for stroke?

Do you prefer your doctor and hospital incompetence NOT KNOWING? OR NOT DOING?

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

---

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.

Memory Loss Associated With Alzheimer's May Be Preventable And Reversible

Well shit, parts of this have been known for years. Is our fucking medical establishment that incompetent that they don't bother to read research? Not only do I have to do the work of the stroke medical establishment I also have to take on the dementia establishment.

Curcumin Loaded Nanoparticles Potently Induce Adult Neurogenesis and Reverse Cognitive Deficits in Alzheimer’s Disease Model via Canonical Wnt/β-catenin Pathway  Dec. 2013 

Effect of canonical Wnt inhibition in the neurogenic cortex, hippocampus, and premigratory dentate gyrus progenitor pool  Dkk1 expression June 2013 

Loss of Dickkopf-1 Restores Neurogenesis in Old Age and Counteracts Cognitive Decline  Feb. 2013 

Really nothing new in this latest research:

Memory Loss Associated With Alzheimer's May Be Preventable And Reversible

Alzheimer’s disease routinely appears at or near the top of diseases most feared by Americans. It’s terrifying to contemplate and devastating when it happens. The memory loss that is a common symptom of Alzheimer’s begins with loss of function at the synaptic connections between neurons and progresses to the destruction of the neurons themselves. Research carried out by a team at University College London and recently reported in the journal Current Biology details how molecular changes at the synapse damage memory and suggests ways that the memory loss associated with Alzheimer’s may be prevented and possibly reversed.

The Wnt and Dkk1 proteins
A protein called Wnt plays an important role in a process called canonical Wnt signaling that is involved in the formation and development of synapses. Wynt signaling is also involved in modulating communication between one neuron and the next that takes place at the synapse. Another type of protein called Dkk1 (Dickkopf-1) can block Wnt signaling by binding to the Wnt receptors in a neuron.
Prior research suggests that reductions in Wnt signaling and increases in Dkk1 are involved in Alzheimer’s. For example, the beta ameloid deposits that play a critical role in Alzheimer’s appear to increase production of Dkk1 in the brain, and post-mortem examinations of the brains of advanced Alzheimer’s patients show elevated levels of Dkk1. While these and other findings are suggestive, the effects on memory of the interplay between Wnt and Dkk1 at the synapse have not been fully examined.
The research team examined the effects of elevated levels of Dkk1 in the hippocampus (a structure in the brain that plays an important role in long-term memory) of specially bred mice. The mice were genetically engineered to increase secretion of Dkk1 when they are injected with the antibiotic doxycycline. Administration of doxycycline thus served as an on-off switch for producing elevated levels of Dkk1 in the hippocampus.

One group of specially bred mice (the Dkk1 mice) were given doxycycline injections for two weeks prior to testing while a group of their littermates (the control mice) were not. The researchers then carried out a variety of tests to determine how elevated levels of Dkk1 affected activity at the synapse and how blocking Wnt with Dkk1 affected long-term memory.
Examination of the effects of Dkk1 at the synapse showed that elevated levels of Dkk1 reduced the ability of a sending neuron to excite or activate a receiving neuron. Dkk1 also reduced the ability of two neurons to form stronger connections after recent co-activation (that is, Dkk1 reduced neural plasticity). In addition, the researchers were able to identify the specific metabolic pathways through which Dkk1 causes loss of function at the synapse.

A Morris water maze test. Credit: Samueljohn.de – Own work, CC BY-SA 3.0/Wikipedia Commons

Wnt, Dkk1 and memory loss
How blocking Wnt with Dkk1 affects memory was examined with water maze and conditioned fear tests. In a water maze test a mouse is placed in a circular tank of water that has a submerged platform just under the surface. The mouse has to swim until it finds the platform. Memory is measured in terms of how long it takes the mouse to swim back to the platform on later trials after it has found the platform on the first trial. The Dkk1 mice took significantly longer than the control mice to return to the platform on the third and fourth trials indicating that preventing Wnt from acting with Dkk1 disrupted spatial memory.
In the conditioned fear test, the mice were placed in an environment that was new to them and a shock was administered through an electrified grid in the floor. If the mouse remembers being shocked, it will be afraid the next time it’s placed in the same environment. When a mouse is afraid, it will freeze (cease all movement other than breathing). The length of time spent frozen is taken as a measure of the mouse’s memory for having been shocked the last time it was in the environment.
The mice in the experiment were put back in the environment where they had been shocked 24 hours after the shock had been administered. The control mice froze for roughly twice as long as the Dkk1 mice. This indicates that blocking Wnt activity with Dkk1 disrupts memory associations between locations and painful experiences that happened in those locations.
Taken together, the water maze and conditioned fear tests indicate that Wnt signaling plays an important role in the formation of memories and that blocking Wnt signaling with Dkk1 in the hippocampus can produce long-term memory deficits.
Can the loss of memory function be reversed?
Early-stage Alzheimer’s is typically diagnosed after synapses have been damaged but before there has been significant destruction of neurons. Can synaptic function be restored by eliminating the Dkk1 block on Wnt signaling or has Dkk1 blocking permanently damaged the synapses so that function is permanently lost?
The researchers addressed this question with new groups of Dkk1 and control mice. The Dkk1 mice were given doxycycline injections for two weeks to elevate levels of Dkk1 in the hippocampus. This was followed by two weeks without doxycycline to bring Dkk1 back to normal levels.
The Dkk1 and control mice were compared after the four-week, on-off Dkk1 period. The ability of sending neurons to excite receiving neurons was not significantly different between the two groups of mice. In other words, synaptic excitability was completely restored in the Dkk1 mice. The Dkk1 mice also exhibited full restoration of neural plasticity.
The mice were also given the conditioned fear test after the four-week, on-off period. Freezing time was the same for the Dkk1 mice and the controls. The long-term memory function that was lost when Wnt signaling was blocked by Dkk1 was restored when Dkk1 levels were reduced and Wnt was again able to function normally.
The restoration of long-term memory function combined with the restoration of excitation and neural plasticity at the synapse that was observed in these experiments indicates that the memory deficits that are symptomatic of Alzheimer’s disease may be reversible if the Dkk1 block on Wnt signaling can be removed before the destruction of neurons takes place.

Curcumin Loaded Nanoparticles Potently Induce Adult Neurogenesis and Reverse Cognitive Deficits in Alzheimer’s Disease Model via Canonical Wnt/β-catenin Pathway

I would think that increasing the cognitive ability of survivors would help tremendously in their recovery. How long before your doctor puts this into practice? Never? Its going to be impossible for you to get curcumin nanoparticles on your own.
http://pubs.acs.org/doi/abs/10.1021/nn405077y

Abstract

Neurogenesis, a process of generation of new neurons, is reported to be reduced in several neurodegenerative disorders including Alzheimer’s disease (AD). Induction of neurogenesis by targeting endogenous neural stem cells (NSC) could be a promising therapeutic approach to such diseases by influencing the brain self regenerative capacity. Curcumin, a neuroprotective agent has poor brain bio-availability. Herein, we report that curcumin-encapsulated PLGA nanoparticles (Cur-PLGA-NPs) potently induce NSC proliferation and neuronal differentiation in vitro and in the hippocampus and subventricular zone of adult rats, as compared to uncoated bulk curcumin. Cur-PLGA-NPs induce neurogenesis by internalization into the hippocampal NSC. Cur-PLGA-NPs significantly increase expression of genes involved in cell proliferation (reelin, nestin and Pax6) and neuronal differentiation (neurogenin, neuroD1, neuregulin, neuroligin and Stat3). Curcumin nanoparticles increase neuronal differentiation by activating the Wnt/β-catenin pathway, involved in regulation of neurogenesis. 

These nanoparticles caused enhanced nuclear translocation of β-catenin, decreased GSK-3β levels, and increased promoter activity of the TCF/LEF and cyclin-D1. Pharmacological and siRNA mediated genetic inhibition of the Wnt pathway blocked neurogenesis stimulating effects of curcumin. These nanoparticles reverse learning and memory impairments in an amyloid beta induced rat model of AD like phenotypes, by inducing neurogenesis. In silico molecular docking studies suggest that curcumin interacts with Wif-1, Dkk and GSK-3β. These results suggest that curcumin nanoparticles induce adult neurogenesis through activation of the canonical Wnt/β-catenin pathway, and may offer a therapeutic approach to treating neurodegenerative diseases such as AD, by enhancing a brain self repair mechanism.

Wnt Activation of Immortalized Brain Endothelial Cells as a Tool for Generating a Standardized Model of the Blood Brain Barrier In Vitro

Another writeup on getting across the blood brain barrier. Ask your doctor and researcher which one they prefer. We are going to need this for drug delivery unless nasal wins out.
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0070233

Abstract

Reproducing the characteristics and the functional responses of the blood–brain barrier (BBB) in vitro represents an important task for the research community, and would be a critical biotechnological breakthrough. Pharmaceutical and biotechnology industries provide strong demand for inexpensive and easy-to-handle in vitro BBB models to screen novel drug candidates. Recently, it was shown that canonical Wnt signaling is responsible for the induction of the BBB properties in the neonatal brain microvasculature in vivo. In the present study, following on from earlier observations, we have developed a novel model of the BBB in vitro that may be suitable for large scale screening assays. This model is based on immortalized endothelial cell lines derived from murine and human brain, with no need for co-culture with astrocytes. To maintain the BBB endothelial cell properties, the cell lines are cultured in the presence of Wnt3a or drugs that stabilize β-catenin, or they are infected with a transcriptionally active form of β-catenin. Upon these treatments, the cell lines maintain expression of BBB-specific markers, which results in elevated transendothelial electrical resistance and reduced cell permeability. Importantly, these properties are retained for several passages in culture, and they can be reproduced and maintained in different laboratories over time. We conclude that the brain-derived endothelial cell lines that we have investigated gain their specialized characteristics upon activation of the canonical Wnt pathway. This model may be thus suitable to test the BBB permeability to chemicals or large molecular weight proteins, transmigration of inflammatory cells, treatments with cytokines, and genetic manipulation.

Effect of canonical Wnt inhibition in the neurogenic cortex, hippocampus, and premigratory dentate gyrus progenitor pool

So we don't want Dickkopf-1 because it reduces neurogenesis. Ask your doctor how to accomplish that. 

Effect of canonical Wnt inhibition in the neurogenic cortex, hippocampus, and premigratory dentate gyrus progenitor pool

Abstract

Canonical Wnt signaling is crucial for the correct development of both cortical and hippocampal structures in the dorsal telencephalon. In this study, we examined the role of the canonical Wnt signaling in the dorsal telencephalon of mouse embryos at defined time periods by inhibition of the pathway with ectopic expression of Dkk1. Transgenic mice with the D6-driven Dkk1 gene exhibited reduced canonical Wnt signaling in the cortex and hippocampus. As a result, all hippocampal fields were reduced in size. Neurogenesis in the dentate gyrus was severely reduced both in the premigratory and migratory progenitor pool. The lower number of progenitors in the dentate gyrus was not rescued after migration to the subgranular zone and thus the dentate gyrus lacked the entire internal blade and a part of the external blade from postnatal to adult stages.

Secreted Frizzled-Related Protein 3 Regulates Activity-Dependent Adult Hippocampal Neurogenesis

So we don't want SFRP3 because it reduces neurogenesis. Ask your doctor how to accomplish that.
http://www.cell.com/cell-stem-cell/retrieve/pii/S1934590912007035

• Highlights
• Wnt inhibitor sFRP3 exhibits activity-dependent expression in the adult hippocampus
• sFRP3 maintains quiescence of adult hippocampal radial glia-like neural stem cells
• sFRP3 inhibits maturation, dendritic development, and spinal formation of new neurons
• sFRP3 partially mediates activity-dependent adult hippocampal neurogenesis

Summary

Adult neurogenesis, the process of generating mature neurons from adult neural stem cells, proceeds concurrently with ongoing neuronal circuit activity and is modulated by various physiological and pathological stimuli. The niche mechanism underlying the activity-dependent regulation of the sequential steps of adult neurogenesis remains largely unknown. Here, we report that neuronal activity decreases the expression of secreted frizzled-related protein 3 (sFRP3), a naturally secreted Wnt inhibitor highly expressed by adult dentate gyrus granule neurons. Sfrp3 deletion activates quiescent radial neural stem cells and promotes newborn neuron maturation, dendritic growth, and dendritic spine formation in the adult mouse hippocampus. Furthermore, sfrp3 reduction is essential for activity-induced adult neural progenitor proliferation and the acceleration of new neuron development. Our study identifies sFRP3 as an inhibitory niche factor from local mature dentate granule neurons that regulates multiple phases of adult hippocampal neurogenesis and suggests an interesting activity-dependent mechanism governing adult neurogenesis via the acute release of tonic inhibition.

Loss of Dickkopf-1 Restores Neurogenesis in Old Age and Counteracts Cognitive Decline

Your doctor should be able to take this and create a stroke protocol for you. But your incompetent? doctor will DO NOTHING! 

Loss of Dickkopf-1 Restores Neurogenesis in Old Age and Counteracts Cognitive Decline

Summary

Memory impairment has been associated with age-related decline in adult hippocampal neurogenesis. Although Notch, bone morphogenetic protein, and Wnt signaling pathways are known to regulate multiple aspects of adult neural stem cell function, the molecular basis of declining neurogenesis in the aging hippocampus remains unknown. Here, we show that expression of the Wnt antagonist Dickkopf-1 (Dkk1) increases with age and that its loss enhances neurogenesis in the hippocampus. Neural progenitors with inducible loss of Dkk1 increase their Wnt activity, which leads to enhanced self-renewal and increased generation of immature neurons. This Wnt-expanded progeny subsequently matures into glutamatergic granule neurons with increased dendritic complexity. As a result, mice deficient in Dkk1 exhibit enhanced spatial working memory and memory consolidation and also show improvements in affective behavior. Taken together, our findings show that upregulating Wnt signaling by reducing Dkk1 expression can counteract age-related decrease in neurogenesis and its associated cognitive decline.

Radial glial neural progenitors regulate nascent brain vascular network stabilization via inhibition of wnt signaling.

We need new blood vessels to support our neurogenesis and possibly stem cells. Your doctor should be able to decipher this into a stroke protocol.  Lots of images at the link.
http://www.ncbi.nlm.nih.gov/pubmed/23349620

Abstract

The cerebral cortex performs complex cognitive functions at the expense of tremendous energy consumption. Blood vessels in the brain are known to form stereotypic patterns that facilitate efficient oxygen and nutrient delivery. Yet little is known about how vessel development in the brain is normally regulated. Radial glial neural progenitors are well known for their central role in orchestrating brain neurogenesis. Here we show that, in the late embryonic cortex, radial glial neural progenitors also play a key role in brain angiogenesis, by interacting with nascent blood vessels and regulating vessel stabilization via modulation of canonical Wnt signaling. We find that ablation of radial glia results in vessel regression, concomitant with ectopic activation of Wnt signaling in endothelial cells. Direct activation of Wnt signaling also results in similar vessel regression, while attenuation of Wnt signaling substantially suppresses regression. Radial glial ablation and ectopic Wnt pathway activation leads to elevated endothelial expression of matrix metalloproteinases, while inhibition of metalloproteinase activity significantly suppresses vessel regression. These results thus reveal a previously unrecognized role of radial glial progenitors in stabilizing nascent brain vascular network and provide novel insights into the molecular cascades through which target neural tissues regulate vessel stabilization and patterning during development and throughout life.

Full article here:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3551952/

Molecular Control of Axon Branching

We need this so ask your doctor for how to do this to help your recovery. That's after they explain the damage to your white matter. 

Molecular Control of Axon Branching

Abstract

Axon branching is a complex morphological process, the regulation of which we are just beginning to understand. Many factors known to be important for axon growth and guidance have emerged as key regulators of axon branching. The extrinsic factors implicated in axon branching include traditional axon guidance cues such as the slits, semaphorins, and ephrins; neurotrophins such as BDNF; the secreted glycoprotein Wnt; the extracellular matrix protein anosmin-1; and certain transmembrane cell adhesion molecules—as well as sensory experience and neuronal activity. Although less is known about the intracellular control of axon branching, in recent years significant advances have been made in this area. Kinases and their regulators, Rho GTPases and their regulators, transcription factors, ubiquitin ligases, and several microtubule and actin-binding proteins are now implicated in the control of axon branching. It is likely that many more branching regulators remain to be discovered, as do the links between extrinsic cues and intracellular signaling proteins in the control of axon branching.

Signaling mechanisms regulating adult neural stem cells and neurogenesis

I only put this out here because of the sonic hedgehog. You'll have to get your doctor to buy the full article.
 http://www.sciencedirect.com/science/article/pii/S0304416512002577

Abstract

Background

Adult neurogenesis occurs throughout life in discrete regions of the mammalian brain and is tightly regulated via both extrinsic environmental influences and intrinsic genetic factors. In recent years, several crucial signaling pathways have been identified in regulating self-renewal, proliferation, and differentiation of neural stem cells, as well as migration and functional integration of developing neurons in the adult brain.

Scope of review

Here we review our current understanding of signaling mechanisms, including Wnt, notch, sonic hedgehog, growth and neurotrophic factors, bone morphogenetic proteins, neurotransmitters, transcription factors, and epigenetic modulators, and crosstalk between these signaling pathways in the regulation of adult neurogenesis. We also highlight emerging principles in the vastly growing field of adult neural stem cell biology and neural plasticity.

Major conclusions

Recent methodological advances have enabled the field to identify signaling mechanisms that fine-tune and coordinate neurogenesis in the adult brain, leading to a better characterization of both cell-intrinsic and environmental cues defining the neurogenic niche. Significant questions related to niche cell identity and underlying regulatory mechanisms remain to be fully addressed and will be the focus of future studies.

General significance

A full understanding of the role and function of individual signaling pathways in regulating neural stem cells and generation and integration of newborn neurons in the adult brain may lead to targeted new therapies for neurological diseases in humans. This article is part of a Special Issue entitled Biochemistry of Stem Cells.

---

Highlights

► Adult neurogenesis is regulated via both extrinsic environmental influences and intrinsic genetic factors. ► We review individual signaling mechanisms and their cross-talk in regulating adult neurogenesis. ► We highlight emerging principles in the growing field of adult neural stem cell biology.

So after your doctor has digested this article, ask him/her how they are going to use it to help your neurogenesis along.  Don't be afraid, they'll probably just ask if you want one for a pet.