酪氨酸蛋白激酶Src和Fyn对脑缺血再灌注大鼠海马CA_1区神经细胞损伤的影响

Effect of tyrosine protein kinase Src and Fyn on transient brain ischemia/reperfusion-induced neuronal death in rat hippocampus

目的观察胞内可溶性酪氨酸蛋白激酶Src、Fyn蛋白对脑缺血再灌注后海马CA1区神经细胞存活的影响。方法采用Pulsinelli-Brierley4动脉阻塞(4-VO)大鼠全脑缺血模型,缺血前连续3d脑室注射Src、Fyn反义寡核苷酸(antisense oligodeoxy-nucletides,AS ODNs)抑制Src、Fyn蛋白的表达后缺血,复灌5d,石蜡切片,焦油紫染色,图像分析测定单位面积内焦油紫染色细胞面积总和,与缺血组及错义寡核苷酸组(missense oligodeoxy-nucletides,MS ODNs)比较进行形态学分析。结果Src、Fyn反义寡核苷酸对脑缺血再灌注后海马CA1神经细胞均有明显保护作用,以Src AS ODNs保护作用更明显,与缺血组比较约有50%细胞存活(P<0.05)。结论胞内可溶性酪氨酸蛋白激酶Src、Fyn蛋白的含量降低对脑缺血再灌注后海马神经细胞有明显保护作用,且以Src作用更为明显。

Objective To investigate whether the tyrosine protein kinase Src and Fyn were involved in the neuronal cell death induced by cerebral ischemia followed by reperfusion. Methods Transient （15min） brain ischemia was induced by the four-vessel occlusion in Sprague Dawleyrats. The antisense oligodeoxynucleotides （ODNs） of Src and Fyn were used to suppress the expression of Src and Fyn by intracerebroventricular infusion once per day for 3 days before ischemia. After 5 days reperfusion, rats were perfusion-fixed with paraformaldehyde and cresyl violet staining were used to examine the survival of CA1 pyramidal cells of hippocampus. Results Both Sre and Fyn antisense ODNs showed a protective role against neuronal cell death induced by cerebral ischemia/reperfusion. Furthermore ,administration of Sre antisense ODNs had more protective effect on the neuronal death than Fyn antisense ODNs. Conclusions Both Src and Fyn play an important role in neuronal death induced by cerebral ischemia followed by reperfusion, while Sre dominate over Fyn in the process.