Abstract P70: SPAK/OSR1 Signaling as a Novel Target for Post-Stroke Oxidative Stress Brain Injury

 So cultured mouse neurons were used. Your hospital is responsible for getting live mouse testing done and then human testing. Then distribute the created protocols worldwide.

Abstract P70: SPAK/OSR1 Signaling as a Novel Target for Post-Stroke Oxidative Stress Brain Injury

Jun Wang

,

Victoria M Fiesler

,

Gulnaz Begum

,

Mohammad Iqbal H Bhuiyan

,

Shuying Dong

,

Eric Li

,

Kristopher T Kahle

,

Jinwei Zhang

,

Xianming Deng

,

Yan Yin

,

Dandan Sun

Originally published11 Mar 2021https://doi.org/10.1161/str.52.suppl_1.P70Stroke. 2021;52:AP70

Abstract

Stroke is the second leading cause of death worldwide, and ischemic stroke accounts for the vast majority of stroke cases. Currently, recombinant tissue plasminogen activator and endovascular thrombectomy are the two primary therapy strategies for acute ischemic stroke patients. Reactive oxygen species (ROS)-mediated oxidative stress can cause brain injury during reperfusion. We have shown that the Ste20/SPS1-related proline-alanine-rich protein kinase/oxidative stress-responsive kinase-1 (SPAK/OSR1) are activated in ischemic stroke brains, resulting in worsened outcomes in murine stroke models. SPAK activation induces the production of pro-inflammatory cytokines. Post-stroke administration of a novel SPAK inhibitor ZT-1a attenuates cerebral edema and protects against brain damage in in vivo model of ischemic stroke. However, whether ROS mediated oxidative stress directly activate SPAK/OSR1 pathway and induces SPAK pro-inflammatory cytokine production in ischemic brains remains unknown. In our extended study, we examined activation of SPAK/OSR1 and its substrate Na-K-Cl cotransporter (NKCC1) in cultured mouse primary neurons in response to hydrogen peroxide (H₂O₂)-mediated oxidative stress. We found that exposure of neurons to H₂O₂ for 24 hrs triggered upregulation of protein expression and phosphorylation activation of SPAK/OSR1 and NKCC1 (p < 0.05), which are accompanied with an increase in intracellular Na⁺ concentration and neuronal death (p < 0.01). These changes were blocked by an ROS scavenger ebselen. Interestingly, both novel SPAK inhibitor ZT-1a and NKCC1 inhibitor bumetanide are able to block H₂O₂-induced neuronal damage. We are in the process to assess effects of SPAK inhibitor ZT-1a in reducing ROS-mediated inflammation and brain injury in in vivo model of ischemic stroke. Together, our study suggests that ROS can activate SPAK/OSR1 complex during reperfusion injury and the therapeutic potentials of SPAK inhibitor ZT-1a for ischemic stroke.

 

New review examines potential of antioxidant therapies to combat neurodegenerative disorders

Would anything here help stroke recovery?
http://www.news-medical.net/news/20160802/New-review-examines-potential-of-antioxidant-therapies-to-combat-neurodegenerative-disorders.aspx

A new review examines the potential of antioxidant approaches for the treatment of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and multiple sclerosis.
Certain compounds that are involved in oxidative stress look like promising therapeutic targets. For example, researchers are investigating the potential of increasing antioxidant capacity by targeting what's known as the Nrf2 pathway, as well as developing inhibitors of NADPH oxidases, which are key sources of reactive oxygen species. Other potential strategies for limiting oxidative stress in neurodegenerative diseases include reducing the production of nitric oxide, or preventing mitochondrial dysfunction.
"There are still several gaps in our understanding of the basis of oxidative damage in neurodegenerative disorders; however, it is increasingly accepted that many diseases share common pathways of oxidative stress-related damage, and it's likely that significant progress will be made in the design and implementation of effective therapeutic strategies in the next few years," said Dr. Gethin McBean, lead author of the British Journal of Pharmacology review.

Source:

Wiley

Astrocytes can protect brain tissue and reduce disability due to stroke

How long to get something like this into a stroke protocol? Demand an answer from your doctor, it may be useful for you also.
http://www.news-medical.net/news/20130724/Astrocytes-can-protect-brain-tissue-and-reduce-disability-due-to-stroke.aspx

One of regenerative medicine's greatest goals is to develop new treatments for stroke. So far, stem cell research for the disease has focused on developing therapeutic neurons - the primary movers of electrical impulses in the brain - to repair tissue damaged when oxygen to the brain is limited by a blood clot or break in a vessel. New UC Davis research, however, shows that other cells may be better suited for the task.
Published July 23 in the journal Nature Communications, the large, collaborative study found that astrocytes - neural cells that transport key nutrients and form the blood-brain barrier - can protect brain tissue and reduce disability due to stroke and other ischemic brain disorders.
"Astrocytes are often considered just 'housekeeping' cells because of their supportive roles to neurons, but they're actually much more sophisticated," said Wenbin Deng, associate professor of biochemistry and molecular medicine at UC Davis and senior author of the study. "They are critical to several brain functions and are believed to protect neurons from injury and death. They are not excitable cells like neurons and are easier to harness. We wanted to explore their potential in treating neurological disorders, beginning with stroke."
Deng added that the therapeutic potential of astrocytes has not been investigated in this context, since making them at the purity levels necessary for stem cell therapies is challenging. In addition, the specific types of astrocytes linked with protecting and repairing brain injuries were not well understood.
The team began by using a transcription factor (a protein that turns on genes) known as Olig2 to differentiate human embryonic stem cells into astrocytes. This approach generated a previously undiscovered type of astrocyte called Olig2PC-Astros. More importantly, it produced those astrocytes at almost 100 percent purity.
The researchers then compared the effects of Olig2PC-Astros, another type of astrocyte called NPC-Astros and no treatment whatsoever on three groups of rats with ischemic brain injuries. The rats transplanted with Olig2PC-Astros experienced superior neuroprotection together with higher levels of brain-derived neurotrophic factor (BDNF), a protein associated with nerve growth and survival. The rats transplanted with NPC-Astros or that received no treatment showed much higher levels of neuronal loss.
To determine whether the astrocytes impacted behavior, the researchers used a water maze to measure the rats' learning and memory. In the maze, the rats were required to use memory rather than vision to reach a destination. When tested 14 days after transplantation, the rats receiving Olig2PC-Astros navigated the maze in significantly less time than the rats that received NPC-Astros or no treatment.
The investigators used cell culture experiments to determine whether the astrocytes could protect neurons from oxidative stress, which plays a significant role in brain injury following stroke. They exposed neurons co-cultured with both types of astrocytes to hydrogen peroxide to replicate oxidative stress. They found that, while both types of astrocytes provided protection, the Olig2PC-Astros had greater antioxidant effects. Further investigation showed that the Olig2PC-Astros had higher levels of the protein Nrf2, which increased antioxidant activity in the mouse neurons.
"We were surprised and delighted to find that the Olig2PC-Astros protected neurons from oxidative stress in addition to rebuilding the neural circuits that improved learning and memory," said Deng.

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Mn (III) tetrakis (4-Benzoic Acid) porphyrin scavenges reactive species, reduces oxidative stres s, and improves functional recovery after experimental spinal cord injury in rats: comparison with methylprednisolone

Ask your doctor why this wouldn't help in stroke recovery. And then ask when s/he is going to start clinical trials on this.
http://www.biomedcentral.com/content/pdf/1471-2202-14-23.pdf

Abstract

Background

Substantial experimental evidence supports that reactive species mediate secondary damage

after traumatic spinal cord injury (SCI) by inducing oxidative stress. Removal of reactive

species may reduce secondary damage following SCI. This study explored the effectiveness

of a catalytic antioxidant - Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) - in

removing reactive oxygen species (ROS), reducing oxidative stress, and improving functional

recovery in vivo in a rat impact SCI model. The efficiency of MnTBAP was also compared

with that of methylprednisolone – the only drug used clinically in treating acute S

CI.

Results

In vivo measurements of time courses of ROS production by microdialysi

s and microcannula sampling in MnTBAP, methylprednisolone, and saline (as vehicle contr

ol)-treated SCI rats showed that both agents significantly reduced the production of hydrogen

peroxide, but only MnTBAP significantly reduced superoxide elevation after SCI.

In vitro experiments further demonstrated that MnTBAP scavenged both of the preceding ROS, whereas methylprednisolone had no effect on either. By counting the immuno-positive

neurons in the spinal cord sections immunohistochemically stained with anti-nitrotyrosine and anti-4-hydroxy-nonenal antibodies as the markers of protein nitration and membrane lipid peroxidation, we demonstrated that MnTBAP significantly reduced the numbers of 4-hydroxy-nonenal-positive and nitrotyrosine-positive neurons in the sections at 1.55 to 2.55 mm and 1.1 to 3.1 mm, respectively, rostral to the injury epicenter compared to the vehicle-treated animals. By behavioral tests (open field and inclined plane tests), we demonstrated that at 4 hours post-SCI treatment with MnTBAP and the standard met hylprednisolone regimen both significantly increased test scores compared to thos

e produced by vehicle treatment. However, the outcomes for MnTBAP-treated rats were significantly better than those for methylprednisolone-treated animals.

Conclusions

This study demonstrated for the first time in vivo and in vitro that MnTBAP significantly reduced the levels of SCI-elevated ROS and that MnTBAP is superior to methylprednisolone in removing ROS. Removal of ROS by MnTBAP significantly reduced protein nitration and membrane lipid peroxidation in neurons. MnTBAP more effectively reduced neurological deficits than did by methylprednisolone after SCI - the first most important criterion for assessing SCI treatments. These results support the therapeutic potential of MnTBAP in treating SCI.

Effect of dietary olive leaf extract on brain cholesterol, cholesterol ester and triglyceride levels and of brain edema in rat stroke model

It does require pretreatment prior to your stroke but have your researcher study the hyperacute affects
http://rjms.tums.ac.ir/browse.php?a_id=2345&sid=1&slc_lang=en
Article abstract:
  Background : Brain injury by transient complete global brain ischemia (cardiac arrest) and regional incomplete brain ischemia (ischemic stroke) afflicts a very large number of patients with death or permanent disability. Recent studies suggest that olive extracts suppress inflammation and reduce stress oxidative injury.
  Methods: The aim of the study was to evaluate the effect of dietary Olive Leaf Extract (OLE) on brain cholesterol, cholesterol ester and triglyceride levels as well as brain edema in rat stroke model.
  Five groups, each consisting of 12 male Wistar rats, were studied. First and second groups (control and sham) received distilled water, while three treatment groups received oral olive leaf extract (OLE) for 30 days (50, 75 and 100 mg/kg/day, respectively). Two hours after the last dose, each main group was subdivided to Middle Cerebral Artery Occlusion (MCAO)-operated (n=6) and intact subgroups (n=6) for assessment of neuropathology (brain edema) and brain lipid analysis.
  Results: The brain cholesterol, cholesterol ester and triglyceride levels were greater in experimental groups when compared to controls. Olive leaf extracts reduced brain edema in experimental groups of 75 and 100 mg/kg/day.
  Conclusion: Our data suggest that OLE may be cerebroprotective in a rat model of ischemia-reperfusion. Further work is required to extend these observations.