双苯氟嗪对大鼠脑缺血再灌注诱导的海马神经元凋亡的影响

Effects of Dipfluzine on Hippocampal Neuron Apoptosis in Rats Induced by Global Cerebral Ischemia-Reperfusion

目的观察双苯氟嗪(dipfluzine,Dip)对大鼠脑缺血再灌注损伤的抗凋亡作用。方法采用改良的Pulsinelli四动脉结扎法制备大鼠全脑缺血15 min再灌注模型。将大鼠分为两大组,第一大组分为假手术组,缺血15 min再灌注4,8,24及72 h组。第二大组将大鼠分为假手术组、缺血再灌24 h组、溶剂对照组、Dip(0.5,1和2 mg.kg-1)治疗组及氟桂利嗪(flunarizine,Flu)1mg.kg-1阳性对照组。于缺血15 min再灌注开始时尾静脉注射给药。各组均以流式细胞仪测定海马区神经元凋亡率、caspase-3表达量,以及HE染色计数海马CA1区正常神经元密度。结果缺血后,随着再灌时间的延长,海马神经元凋亡率和caspase-3表达量均显著增高,海马CA1区正常神经元密度明显下降,并均于再灌后24 h达显著性改变。给予3个剂量的Dip和Flu均可明显降低海马神经元凋亡率和caspase-3表达量,而Dip 1,2 mg.kg-1和Flu可抑制正常神经元的丢失。结论Dip可明显减轻缺血再灌注损伤,并有明显的抗凋亡作用,其作用机制可能与其抑制钙超载,从而抑制caspase-3的激活有关。

OBJECTIVE To observe the effects of dipfluzine （Dip） on cell apoptosis in rats after global cerebral ischemia-reperthsion injury.METHODS Transient global ischemia model （15 min ischemia followed by 24 h reperfusion） was set up using Pusincllis four-vessel occlusion method. Rats were randomly divided into two groups.Group Ⅰ ： Rats were randomly divided into 5 subgroups according to the different reperfusion time： sham operation group, I-R 4 h （isehemia 15 rain reperfusion 4 h） group, I-R 8 h, I-R 24 h, and I-R 72 h group. GroupⅡ ： Rats were randomly divided into seven subgroups： sham operation group, solw;nt group, I-R 24 h GROUP , Dip （0.5, 1, 2 mg·kg^-1） group, and Flu 1 mg·kg^-1 group. Drugs were given by caudal vein injection when starting repeffusion, The hippocampal neuron apoptosis rate and the expression of caspase-3 were identified by FCM＋ And the density of normal pyramidalcells in hippocampai CAI sector was measured by HE staining. RESULTS With the prolongation of repcffusion lime, the hippncampal neuron apnptosis tale and expression of easpasc-3 were otwiously increased, and the number of normal pyramidal cells was decreased. All these indicators showed signifieant changes by the repeffusion 24 h after a 15 min of global ischemia. And three doses of Dip decreased the apoptosis rate and expression of caspase-3. Dip 1mg·kg^-1 and Dip 2mg·kg^-1 prevented from the lost of normal neuron. CONCLUSION Dip can prevent from the ischemia-repeffusion injury and possess Ihe antiapoptotic effect on cerebral neurons, Its mechanism may be relate to its inhibition of eakcium overload and then inhibit the activation of castile-3.