Page 91 - JCTR-11-4

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

within the hemispheres nor between the hemispheres 4. Discussion
within treatment groups (Figure 6B). Similarly, the
percentage of NeuN /BrdU co-expressing cells (an The present study investigated the sustained impact of
+
+
assessment of acute phase neurogenesis) revealed no acute pharmacological inhibition of NADPH oxidase
significant interaction between hemisphere and treatment using VAS2870 following experimentally induced stroke.
Within the first 48 h following post-stroke VAS2870
(F [1,4] = 1.60, p=0.27), nor any variation between
treatment groups (F [1,4] = 0.776, p=0.43) or between administration, we demonstrated an improvement in
hemispheres (F [1,4] = 0.396, p=0.56). Post hoc analysis survival probability. However, this survival benefit was
confirmed no significant differences between groups paradoxically accompanied by a transient worsening of
within the hemispheres nor between the hemispheres neurological function at day 2 post-stroke. Furthermore,
within treatment groups (Figure 6C). despite improved survival, VAS2870 treatment did not
yield significant improvements in other outcomes assessed
3.5. Effects of post-MCAO NADPH oxidase inhibition at later time points. Specifically, we observed no significant
on angiogenesis reduction in infarct volume attributable to VAS2870
treatment at either 48 h or 11 days post-MCAO compared
Angiogenesis, measured as comparative post-MCAO to the vehicle-treated group. Similarly, functional recovery
sub-acute vascular density, was assessed using CD31
expression immunofluorescence staining at day 11 post- and cognitive function showed no significant benefit
of VAS2870 treatment at days 10 and 11. Furthermore,
MCAO. Representative images showing CD31 staining analyses of circulating biomarkers, neurogenesis, and
within contralateral and ipsilateral CTX and STR regions vascular density at day 11 did not demonstrate any benefit
for both vehicle- and VAS2870-treated groups are shown of VAS2870 treatment.
in Figure 7A. Ipsilateral CD31 immunofluorescence,
when normalised to the contralateral hemisphere, showed The enhanced survival following VAS2870 treatment
no significant overall effect of treatment (vehicle vs. during the early post-stroke recovery phase has
VAS2870; F [1,8] = 4, p=0.08), nor significant interaction therapeutic benefit; however, the mechanisms underlying
between treatment and brain region (F [1,8] = 0.003, this improved survival are unclear. Considering the role of
6,15
p=0.96; Figure 7B). Post hoc analysis of vehicle- versus OS in early oedema and BBB breakdown, it is plausible
VAS2870-treated groups revealed no significant difference that VAS2870 attenuates these processes, particularly
in immunofluorescence values within CTX or STR at day as cytotoxic oedema formation is a known contributor
15
11 post-MCAO (Figure 7B). CD31 immunofluorescence to early mortality. Evidence suggests that intravenous
within hemispheres revealed a significant overall interaction administration of VAS2870, given before reperfusion at
between region and treatment within the contralateral 5 h post-MCAO, resulted in reduced BBB permeability,
hemisphere (F [1,8] = 7.00, p=0.03), but not in the ipsilateral NOX2/4 expression, AQP4 expression and oedema at
hemisphere (F [1,8] = 2.89, p=0.13; Figure 7C). No overall 24 h following ischaemia, supporting the potential that
49
effect of treatment was observed within either hemisphere enhanced survival may be related to an attenuation of
(contralateral: F [1,8] = 0.338, p=0.58; ipsilateral: these processes. Alternatively, VAS2870 may help mitigate
F [1,8] = 0.506, p=0.506; Figure 7C). Significant differences early secondary injury events, such as haemorrhagic
between region variation were only observed within the transformation or acute systemic inflammatory responses.
contralateral hemisphere (contralateral: F [1,8] = 10.9, However, this survival benefit did not correspond to
p=0.01; ipsilateral: F [1,8] = 0.593, p=0.46; Figure 7C). improved demonstrable neuroprotection or functional
Post hoc analysis revealed a significant difference in CD31 recovery with VAS2870 treatment. Functional outcome
expression between CTX and STR regions within the worsened during the acute recovery phase following
contralateral hemisphere of the vehicle-treated group VAS2870 treatment, although it did not cause weight loss,
(contralateral: p=0.006; ipsilateral: p=0.88). No differences a measure of general well-being. It is possible that the
were seen between regions in the VAS2870-treated group VAS2870 dose or administration time point, while effective
(contralateral: p=0.88; ipsilateral: p=0.78; Figure 7C). No in improving acute survival, was insufficient to provide
within-region difference was observed when comparing substantial protection against cell death or to promote
vehicle- to VAS2870-treated groups within the contralateral functional repair pathways. The pathways influencing early
(CTX: p=0.09; STR: p=0.98) or ipsilateral hemispheres survival may be distinct from those determining tissue
(CTX: p=0.97; STR: p=0.47; Figure 7C). Additional post hoc salvage and long-term recovery in this model. It is possible
analysis revealed no significant differences within treatment that post-stroke VAS2870 administration, while sufficient
groups and regions between contralateral and ipsilateral to impact acute mortality, was too late to salvage significant
hemispheres (Figure 7C). penumbral tissue compared to interventions initiated

Volume 11 Issue 4 (2025) 85 doi: 10.36922/jctr.25.00018