| Objective:This study uses the SD rat model of cerebral ischemia and reperfusion injury to evaluate the neuroprotective efficacy of Salvianolate and its mechanisms through the infract volume and morphological changes and the amount of apoptosis cells in brain tissues as well as Hsp22, P-AKT, P-GSK3beta, AKT and GSK3beta proteins expression changes.Methods:60healthy male SD rats were randomly divided into three major groups:sham operation group, ischemia/reperfusion model group, the Salvianolate intervention group. Ischemic/reperfusion model group and the Salvianolate intervention group were sub-divided into the cerebral infarction group, the apoptosis cells group, the H&E group and the Western Blot group. The middle cerebral artery occlusion was used to perform the rat cerebral ischemic-reperfusion injury model, and then the animals were lumbar injected by Salvianolate (18mg/kg, QD) at Oh,24h and48h after reperfusion. The animals were killed and samples of brain tissue which were collected immediately at the72hours after reperfusion. The neurological deficit score was performed by another investigator and infarct volume was stained with TTC. The morphological changes of brain tissue were detected by H&E staining. Cellular apoptosis were measured using the TUNEL staining. The expression changes of HSP22in rat brain tissue detected by Western Blot, as well as P-AKT, P-GSK3β, AKT and GSK3β expression. Data were analyzed by software of SPSS18.0.Result:1. The neurological deficit score of ischemic-reperfusion group was the highest among three experimental groups, the intervention group was significantly lower than the ischemic-reperfusion model group (p<0.05) but still higher than the sham group. TTC staining showed no infract area in sham group, infract area was observed in model group and intervention group. A significant reduction of infract area was appeared after treatment with Salvianolate compared with ischemic-reperfusion model group.2. TUNEL staining in sham group only showed a very small number of TUNEL-positive cells. The number of TUNEL-positive cells in intervention group was significantly less than model group(P<0.05).3. In sham group, the brain tissue showed normal structure by H&E staining. In ischemic-reperfusion model group, nerve cells were sparse and packed irregular, nuclear condensation and fragmentation. The pathological changes were decreased after intervening with Salvianolate.4. Western Blot data showed us that comparing with sham group, the expression of HSP22protein were rise in model group and intervention group, which had the highest level of proteins expression(p<0.05).5. The expression of P-AKT and P-GSK3β proteins were rise in model group and intervention group, and the intervention group showed the highest level of expression of P-AKT and P-GSK3beta, but the expression of AKT and GSK3β showed no obvious difference among the three groups(p>0.05).Conclusion:Treatment with Salvianolate in cerebral ischemic and reperfusion model can significantly increase HSP22expression and AKT activity, inhibit GSK3beta activity, reduce the volume of cerebral infarction, inhibit cell apoptosis and alleviate the pathological changes, reduce brain tissue ischemia-reperfusion injury and exhibit neuroprotective effect. |
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