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

 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。

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

OBJECTIVE To observe the effects of dipfluzine(Dip) on cell apoptosis in rats after global cerebral ischemia-reperfusion injury.METHODS Transient global ischemia model(15 min ischemia followed by 24 h reperfusion) was set up using Pusinellis four-vessel occlusion method.Rats were randomly divided into two groups.GroupⅠ: Rats were randomly divided into 5 subgroups according to the different reperfusion time: sham operation group,I-R 4 h(ischemia 15 min 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,solvent 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 reperfusion.The hippocampal neuron apoptosis rate and the expression of caspase-3 were identified by FCM.And the density of normal pyramidal cells in hippocampal CA₁ sector was measured by HE staining.RESULTS With the prolongation of reperfusion time,the hippocampal neuron apoptosis rate and expression of caspase-3 were obviously increased,and the number of normal pyramidal cells was decreased.All these indicators showed significant changes by the reperfusion 24 h after a 15 min of global ischemia.And three doses of Dip decreased the apoptosis rate and expression of caspase-3.Dip 1 mg·kg^-1 and Dip 2 mg·kg^-1 prevented from the lost of normal neuron.CONCLUSION Dip can prevent from the ischemia-reperfusion injury and possess the antiapoptotic effect on cerebral neurons.Its mechanism may be relate to its inhibition of calcium overload and then inhibit the activation of caspase-3.