Skip to main content
. 2020 Nov 11;10(1):e12024. doi: 10.1002/jev2.12024

FIGURE 6.

FIGURE 6

EV‐induced regulation of autophagy reduces post‐stroke brain injury and improves neurological recovery. (a) Representative immunofluorescence images displaying the biodistribution of ADMSC‐EVs within the ischemic hemisphere. DiI (red spots), NeuN (green) and DAPI (blue). Scale bars, 25 μm. (b‐c) Neuroprotective effects of ADMSC‐EVs in mice exposed to 1 h of middle cerebral artery occlusion (MCAO) followed by 24 h of reperfusion were evaluated by TTC staining. ADMSC‐EVs were injected at the beginning of the reperfusion (EVs 0 h) or at 12 h after reperfusion (EVs 12 h). Mice treated with PBS served as control (n = 8 per group). Quantitative analysis of the infarct size is shown in (c). (d‐e) Mice (n = 10 per condition) were exposed to 1 h of MCAO with subsequent reperfusion for 14 days during which the corner turn test (d) and the tight rope test (e) were performed. Mice received systemic delivery of ADMSC‐EVs or of 3‐MA (15 mg/kg) immediately at the beginning of the reperfusion (EVs 0 h) or 12 h after reperfusion (EVs 12 h). Control mice received PBS only. Motor coordination tests were done at 4, 7, 10 and 14 days after cerebral ischemia. Both EVs 0 h and EVs 12 h groups showed significant improvement in the tight rope test compared to the PBS control group. On day fourteen, only EVs 0 h significantly improved tight rope performance. In the corner turn test, the EVs 0 h and 12 h group showed improvement on day seven, ten and fourteen compared to the PBS control group, and the 3‐MA 12 h group showed improvement on day seven and day ten compared to the PBS control group. (f‐g) The neuronal density was measured in mice treated with PBS (Control), ADMSC‐EVs or of 3‐MA. EVs and 3‐MA were systemically injected at the beginning of the reperfusion or 12 h after reperfusion. NeuN staining within the ischemic lesion site was done on day 14 (n = 10 per group). Scale bars, 50 μm. (g‐h) The autophagic flux was evaluated with BafA1 in MCAO mice that received 3‐MA injection immediately at the beginning of (3‐MA 0 h) or 12 h after reperfusion (3‐MA 12 h). PBS was given to control animals (n = 6 per group). Quantitative analysis of LC3‐II is shown (h). One‐way ANOVA followed by the Tukey's post‐hoc‐test was used, data are shown as mean ± SD. Data are statistically different from each other with *P < 0.05, **P < 0.01, and ***P < 0.001