Page 95 - JCTR-11-4

Journal of Clinical and
Translational Research NADPH oxidase inhibition in a rodent stroke model

relevant factors, such as advanced age, sex differences single-pathway interventions, such as OS inhibition, into
and comorbidities, affecting translatability. Considering broad neuroprotective efficacy, likely due to the inherently
the multifaceted nature of brain ischaemia, this study multifactorial pathophysiology of stroke associated
utilised a monotherapy approach; however, exploration with inflammation, neurodegeneration, alongside OS
of combined VAS2870 treatment with clinically relevant and the modulating influence of factors, such as clinical
ischaemic stroke treatments (e.g., tissue plasminogen comorbidities. This study highlights these challenges
activator) would strengthen clinical relevance. In addition, and underscores the ongoing effort to elucidate and
high infarct volume variability within the MCAO model understand the complex roles of NADPH oxidase enzymes
significantly reduces statistical power, resulting in large in stroke, revealing that while interventions targeting
animal cohorts to detect meaningful differences between them can impact critical survival pathways, achieving
treatment groups. Given the limited data on VAS2870’s comprehensive therapeutic success may require more
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post-stroke safety and efficacy, in addition to animal use nuanced or combined therapies.
ethical considerations, the resource equation was used to
calculate initial sample sizes. This approach allowed for Acknowledgements
the efficient use of resources, considering the uncertain The authors would like to thank Clare Spicer and Dr. David
nature of the success and potential adverse effects of this Watson for their expert technical assistance. We would
novel treatment paradigm. Consequently, the absence also like to acknowledge the support of the Bio-Support
of a priori power analysis is a limitation. Furthermore, Unit, Preclinical Imaging and SLIM Imaging facilities at
high vehicle-treated mortality led to reduced and uneven the University of Nottingham. The authors would also like
groups during later analyses. This differential mortality to acknowledge the BioRender platform used to create
reduced sample sizes and statistical power, increasing the graphical abstract (Trotman-Lucas, M. [2025], https://
the risk of type II error and limiting confidence for sub- BioRender.com/3onrr62).
acute analyses, such as cellular and circulatory marker
levels, due to survivor bias. The reduced 48-h infarct Funding
analysis number, due to MRI technical difficulties and This research was funded by the University of Nottingham’s
uneven sampling, may also indicate sampling bias and Faculty of Science Pump Priming Award (Grant number:
must be considered. It should also be noted that animals A929A2).
reaching humane endpoints, informed by the Ischaemia
Models: procedural refinements of in vivo Experiments Conflict of interest
guidelines, were included as events within survival
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analyses. These animals, due to severe stroke impact, The authors declare that they have no competing interests.
even with extensive supportive care guided by the Author contributions
IMPROVE guidelines, had a low probability of survival
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and, therefore, based on this survival assumption, were Conceptualisation: Melissa Trotman-Lucas, Claire L.
included in the survival analysis. While this captures Gibson, Ulvi Bayraktutan
overall severity, it may influence the interpretation of Investigation: Melissa Trotman-Lucas, Melanie Wood,
outcomes. Finally, expanding outcome assessments to Malcolm J. W. Prior, Jingyuan Ya
include additional acute time points, a broader cytokine Methodology: Melissa Trotman-Lucas, Malcolm J. W. Prior,
panel, direct region-specific tissue analyses (including Jingyuan Ya
hemispheric differences) and a wider variety of functional Writing – original draft: Melissa Trotman-Lucas
assessments would strengthen future investigations. Writing – review & editing: All authors
In summary, this study demonstrates that acute Ethics approval and consent to participate
pharmacological NADPH oxidase inhibition utilising
NOX2/4 inhibitor VAS2870 significantly enhances early This study was conducted in accordance with the UK
survival following experimental stroke in rats. However, Animals (Scientific Procedures) Act, 1986 (Project Licence
this survival benefit was not accompanied by reductions No. PP9645035) and following ethical approval by the
in cerebral damage or improvements in cognitive or University of Nottingham Animal Welfare Ethical Review
functional recovery within the time points assessed. These Body, reported in line with Animal Research Reporting of
findings suggest that while targeting NADPH oxidase- in vivo Experiments (ARRIVE) guidelines.
derived OS with this specific approach impacts survival Consent for publication
pathways, it may not be highly effective as a monotherapy.
These findings highlight the challenge of translating Not applicable.

Volume 11 Issue 4 (2025) 89 doi: 10.36922/jctr.25.00018