SDF-1β does not affect BMSC proliferation.

<p>Colorimetric quantification of DMSO-solubilized MTT formazan at 540 nm showed no differences in proliferation of Tet-Off-SDF-1β compared to Dox-suppressed and Tet-Off-EV controls (1,3, and 7 d, ±100 ng/ml Dox, n = 6, 3 independent experiments).</p

SDF-1β increases the number of surviving BMSCs following exposure to H₂O₂.

<p>Cell number of trypan blue-stained A) Tet-Off-SDF-1β BMSCs and B) Tet-Off-EV control BMSCs after vehicle control or H₂O₂ treatment. SDF-1β significantly increased the number of surviving cells (trypan blue negative) and decreased the number of dying cells (trypan blue positive) in response to H₂O₂ treatment compared to Dox-suppressed and Tet-Off-EV controls (6 h, ±100 ng/ml Dox, ±1.0 mM H₂O₂, ***p<0.0001, −Dox; H₂O₂ vs. +Dox; H₂O₂, n = 3, 3 independent experiments).</p

SDF-1β preserves BMSC morphology following exposure to H₂O₂.

<p>Representative phase contrast micrographs of A) Tet-Off-SDF-1β BMSCs and B) Tet-Off-EV control BMSCs after vehicle control or H₂O₂ treatment. Overexpression of SDF-1β in Tet-Off-SDF-1β BMSCs allows for a greater number of cells with preserved morphology after H₂O₂ treatment relative to Dox-suppressed and Tet-Off-EV controls (6 h, ±100 ng/ml Dox, ±1.0 mM H₂O₂, 20×, 40×, bar 100 µm, n = 3, 3 independent experiments).</p

SDF-1β preserves BMSC nuclear morphology following exposure to H₂O₂.

<p>Representative fluorescence micrographs of Hoechst 33342-stained A) Tet-Off-SDF-1β BMSCs and B) Tet-Off-EV control BMSCs after vehicle control or H₂O₂ treatment. Overexpression of SDF-1β in Tet-Off-SDF-1β BMSCs allows for a greater number of surviving cells and cells with preserved nuclear morphology after H₂O₂ treatment compared to Dox-suppressed and Tet-Off-EV controls (6 h, ±100 ng/ml Dox, ±1.0 mM H₂O₂, 20×, 40×, bar 100 µm, n = 3, 3 independent experiments).</p

Acute eNOS inhibition did not alter early AT1 blocker treatment induced reduction of blood pressure after the induction of cerebral ischemia.

<p>A schematic diagram showing the experimental design (A). Blood pressure telemetry showing the hypotensive effect of Candesartan following cerebral ischemia. This effect was not altered with acute eNOS inhibition before the induction of cerebral ischemia. Data presented as mean±SEM; n = 5–6 animals per group.</p

eNOS inhibition alters nitrosative stress levels after stroke.

<p>Acute L-NIO treatment did not affect the nitrosative stress levels in the ipsilateral hemisphere ()(A). In contrast, L-NIO induced an increased nitrosative stress in the contralateral hemispheres of both candesartan and saline treated animals (F = 4.76, df = 3;p = 0.008)(B). a, b Pairs of means with different letters are significantly different from each other. Data presented as mean±SEM; n = 6–8 per group.</p

eNOS inhibition induces the expression and activity of anti-recovery mechanisms after cerebral ischemia.

<p>Acute L-NIO treatment increased nNOS (F = 7.44, df = 3; p = 0.0014) (A) and Nogo-A expression (B) in the ipsilateral hemisphere (F = 11.68 df = 3; p<0.0001) This increase was associated with a concomitant increase in JNK phosphorylation (F = 28.46, df = 3; p<0.0001) (C). Data presented as mean±SEM. Solid columns represent ipsilateral hemisphere, columns with stripes represent contralateral hemisphere. S refers to the ipsilateral hemisphere; NS refers to the contralateral hemisphere. a, b or A, B Pairs of means with different letters are significantly different from each other. n = 6–8 animals per group.</p

Early AT1 blockade ameliorates ischemia-induced increase in ER stress.

<p>Candesartan treatment at time of reperfusion reduced the expression of CHOP (F = 12.52, df = 3: p = 0.0007) (A), cleaved ATF6 (F = 7.88; df = 3; p = 0.0044) (C) and increased GRP78 expression in an eNOS dependent manner (F = 6.433, df = 3; p = 0.0089)(D). eNOS inhibition increased the expression of full length ATF6 (F = 8.264, df = 3; p = 0.0078) (B) Data presented as mean±SEM. Solid columns represent ipsilateral hemisphere, columns with stripes represent contralateral hemisphere. S refers to the ipsilateral hemisphere; NS refers to the contralateral hemisphere. a, b Pairs of means with different letters are significantly different from each other * n = 4 per group.</p

A schematic representation of the main findings.

<p>Acute administration of ARBs improves short term functional outcome through an eNOS mediated amelioration of ER stress and BDNF/TrkB mediated signaling. Acute eNOS inhibition increases NOGO-A, nNOS expression and JNK phosphorylation.</p

Early AT1 blockade improved 24-hour functional outcome measures while not affecting hemorrhagic transformation in hypertensive animals.

<p>Neither candesartan nor L-NIO had an effect on the development of hemorrhagic transformation (A). Candesartan improved neurological outcome after stroke, as assessed by modified Bederson, in an eNOS independent manner (B). After 7 days, there was a significant effect of time (p<0.01) in all measures of neurobehavioral outcome, demonstrating a gradual recovery from the initial deficits in all treatment groups. The candesartan-treated group (without eNOS inhibition) had better Bederson (2C) and Paw grasp (2D) scores than the other groups, but neither reached statistical significance. a, b: Pairs of means with different letters are significantly different from each other. Data presented as mean±SEM; n = 6–8 per group. Statistical analysis was done as a 2×2 factorial analysis. For clarity, animals that received saline injections before the induction of tMCAO are identified by the treatment they received after tMCAO only.</p