| Objective:The focal cerebral ischemia reperfusion(I/R)model was induced by suture method cause the obstruction of rats,right middle cerebral artery in this research,give hydroxy safflower safflower yellow A(HSYA)in preoperative and postoperative via intravenous intervention respectively.Though observing the neurological defect score,cerebral infarction volume and the influence of HSP70 expression,the apoptosis of nerve cell of brain tissue,after the dose of preoperative and postoperative.Here,we investiggate the preventive and treatment role of HSYA and the effect of expression of HSP70 in middle cerebral artery I/R,and then discuss the molecular mechanism of the neuroprotective of HSYA in cerebral I/R,provide theoretical foundation in the treatment of cerebral infarction.Methods:1.Animal grouping: Seventy healthy male Sprague-Dawley rats weight ing250-300 g were randomly allcated to seven groups(normal control,sham operation,model group,preoperative 5 mg/kg group,preoperative 10 mg/kg group,postoperative 5 mg/kg group,postoperative 10 mg/kg group),10 rats in each group.2.Rat model of transient focal I/R:Facal cerebral ischemia was induced by the right middle cerebral artery occlusion(MCAO)with a nylon monofilament suture.The middle cerebral artery(MCA)was occluded by a suture with an expanded tip from the common carotid artery into the lumen of the internal carotid artery and arrived the origin of the MCA.After 2h of occlusion,the suture was pulled out then reperfusion for 24 h.Sham-operated rats underwent identical surgical procedures without MCAO.3.Drug treatment:The durg(5 mg/kg,10 mg/kg)was respectively administered30 min before and after MACO via tail vein injection in preoperative group and postoperative group.Sham operation and model group rats received same weight placebo(normal saline).4.Measurements: Two hours after waking from Surgery anesthesia,the neurological defect score assessed by the same person.After 24 h repetfusion,all the rats were anesthetized and then decapitated,Take the brains were immediately to detectde the affect of cerebral infarction volume,the expression of HSP70 and the apoptosis of nerve cell in ischemic penumbra.Results:1.Neurological defect score:The prevetment and treatment of HSYA(5mg/kg,10mg/kg)could reduced the neurologgical defect score of rats after ischemia-reperfusion.Compared with model group,the neurologgical defect score of preoperative 10 mg/kg group and postoperative(5mg/kg,10mg/kg)group reduced obviously,the difference was statistically significant(P<0.05).2.Cerebral infarction volume:The prevetment and treatment of HSYA(5mg/kg,10mg/kg)could decreased the cerebral infarction volume of rats after ischemia-reperfusion(P<0.05).Compared with preoperative 5mg/kg group,the cerebral infarction volume of postoperative(5mg/kg,10mg/kg)group significantly decreased,the difference was statistically significant(P<0.05).Compared with preoperative 10 mg/kg group,the cerebral infarction volume of postoperative 10mg/kg group reduced evidently,the difference was statistically significant(P<0.05).3.Immunohistochemical: HSP70 mainly expressed in the cytoplasm,The positive cells expressed rarely in brain tissue of normal control.The expression of positive cells was slightly increased in sham operation(P>0.05).After cerebral I/R,the expression of positive cells in ischemic penumbra was a massive increased,the difference was statistically significant(P<0.01).Compared with model group,the positive cells express of preventment and treatment was significantly reduced,the difference was statistically significant(P<0.01).Compared with preoperative 5mg/kg group,the positive cells express of postoperative(5mg/kg,10mg/kg)group significantly decreased,the difference was statistically significant(P<0.05).Compared with preoperative 10mg/kg group,the positive cells express of postoperative 10mg/kg group significantly decreased,the difference was statistically significant(P<0.01).Compared with postoperative 5mg/kg group,the positive cells express of postoperative 10mg/kg group significantly decreased,the difference was statistically significant(P<0.01).4.Western-boltting:Compared with normal contral,the HSP70 expression of sham operation in ischemic penumbra was significantly increased,the difference was statistically significant(P<0.01).Compared with sham operation,the HSP70 expression of model group in ischemic penumbra was significantly increased,the difference was statistically significant(P<0.01).Compared with preoperative 5mg/kg group,the HSP70 express of postoperative(5mg/kg,10mg/kg)group significantly decreased,the difference was statistically significant(P<0.01).Compared with preoperative 10mg/kg group,the HSP70 express of postoperative 10mg/kg group significantly decreased,the difference was statistically significant(P<0.01).Compared with postoperative 5mg/kg group,the HSP70 express of postoperative 10mg/kg group significantly decreased,the difference was statistically significant(P<0.01).Conclusion:1.The prevetment and treatment of HSYA(5mg/kg,10mg/kg)could decreased the neurologgical defect score after I/R.Compraed with model group,the dose 10mg/kg of HSYA for prevetment and the dose 5mg/kg,10mg/kg for treatment could significantly decreased the neurologgical defect score,improve the limb function defect.2.The prevetment and treatment of HSYA(5mg/kg,10mg/kg)could decreased the cerebral infarction volume after I/R.Under the same dose(5mg/kg,10mg/kg),the treatment dose after I/R is better than the preventive dose before I/R.3.Immunohistochemical and Western-boltting: HSP70 mainly expressed in the cytoplasm,The HSP70 expressed rarely in normal brain tissue,surgical trauma leads to slightly increased the expression of HSP70.After cerebral I/R,the expression of HSP70 in ischemic penumbra was a massive increase.The prevetment and treatment of HSYA both could decreased the expression of ischemic penumbra of Sprague-Dawley mouse after I/R.Under the same dose(5mg/kg,10mg/kg),the treatment dose after I/R is better than the preventive dose before I/R.Compared with5mg/kg dose of HSYA was given after I/R,the 10mg/kg dose after I/R could reduced obviously. |
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