FIGURE 4.

SHY-01 ameliorates infarct area in tMCAO-induced cerebral ischemic stroke. (A) SHY-01 (SHY, 5, 15, or 50 mg/kg) or metformin hydrochloride (Met, 50 mg/kg) were administrated (i.p.) every 24 h after tMCAO, saline was used as vehicle control (Veh), survival rate of rats was recorded 7 days post tMCAO. n = 15 rats/group, Kaplan-Meier analysis followed by a Cox regression test to compare differences between each group. (B) SHY-01 (50 mg/kg) was administrated (i.p.) at 2, 6, or 24 h after reperfusion. The rats were euthanized at 72 h after tMCAO and representative images of brain sections stained with TTC are illustrated. (C) The percentage of the infarct area in whole brain were statistically analyzed, (D) mNSS score in each group were evaluated for each group. Results are expressed as means ± SD. n = 7–8 rats/group for infarct area and mNSS score determination. Statistical analysis was performed using one-way ANOVA with Dunnett’s post hoc tests. ∗p < 0.05, ∗∗p < 0.01, versus the vehicle group. (E) SHY-01 (SHY, 50 mg/kg) or metformin hydrochloride (Met, 50 mg/kg) was administrated (i.p.) at every 24 h after reperfusion, rats were sacrificed at 72 h after tMCAO. Representative images of brain sections stained with TTC were illustrated and (F) percentage of the infarct area in whole brain were statistically analyzed, (G) mNSS score in each group were evaluated as described in material and methods. n = 7–8 rats/group for infarct area and mNSS score determination. Statistical analysis was performed using one-way ANOVA with Dunnett’s post hoc tests. ^∗ p < 0.05, ^∗∗ p < 0.01, versus the vehicle group. ^## p < 0.01, versus metformin hydrochloride group. (H) Immunoblotting of AMPK phosphorylation (Thr 172) level in the contralateral (contra) or ipsilateral (ipsil) of each group and relative phospho-AMPK levels were analyzed using one-way ANOVA with Dunnett’s post hoc tests. ^∗ p < 0.05, ^∗∗ p < 0.01, versus the vehicle group. ^## p < 0.01, versus metformin hydrochloride group.