阿司匹林调控DDR1对大鼠脑缺血再灌注损伤神经细胞凋亡的影响

Effect of Aspirin Regulating DDR1 on Neuronal Apoptosis in Rats with Cerebral Ischemia-reperfusion Injury

目的探究阿司匹林调控盘状结构域受体1(DDR1)对大鼠脑缺血再灌注损伤神经细胞凋亡的影响及作用机制。方法选取30只无特定病原体级6~8周龄雄性SD大鼠,按照随机数字表法分为假手术组、脑缺血再灌注组和阿司匹林干预组,每组10只。线栓法制备脑缺血再灌注模型。观察阿司匹林(60 mg/kg)对神经元细胞凋亡以及胱天蛋白酶3(caspase-3)、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和DDR1蛋白表达的影响。苏木精-伊红染色观察各组大鼠脑神经元形态变化;原位末端转移酶标记技术染色检测神经元细胞凋亡情况。结果与假手术组相比,脑缺血再灌注组海马CA1区域椎体细胞稀疏、有大量空泡,细胞间质增宽,细胞核固缩,神经元细胞凋亡率显著升高[(50.25±8.50)%比(8.98±2.50)%](P<0.01),脑组织caspase-3、Bax和DDR1蛋白表达均显著上调[(2.58±0.38)比(0.71±0.20)、(1.82±0.43)比(0.72±0.27)、(2.33±0.58)比(1.21±0.44)](P<0.05),Bcl-2表达显著下调[(0.61±0.21)比(1.21±0.40)](P<0.05)。与脑缺血再灌注组相比,阿司匹林干预组CA1区域椎体细胞增多、空泡减少,细胞间质变窄,神经元细胞凋亡率降低[(30.25±9.44)%比(50.25±8.50)%](P<0.05),脑组织caspase-3、Bax和DDR1蛋白表达显著下调[(1.47±0.57)比(2.58±0.38)、(1.21±0.31)比(1.82±0.43)、(1.58±0.48)比(2.33±0.58)](P<0.05),Bcl-2表达显著上调[(1.02±0.31)比(0.61±0.21)](P<0.05)。结论阿司匹林能够改善脑缺血再灌注大鼠神经损伤,其作用机制可能与抑制DDR1蛋白表达、减少细胞凋亡相关。

Objective To explore the effect and mechanism of aspirin regulating discoid domain receptor 1(DDR1)on neuronal apoptosis in cerebral ischemia-reperfusion injury in rats.Methods A total of 30 male,6-8 weeks old rats with no specific pathogen were randomly divided into a sham operation group,a cerebral ischemia-reperfusion group and an aspirin intervention group according to random number method,with 10 rats in each group.The cerebral ischemia-reperfusion model was established by suture method.The effects of aspirin(60 mg/kg)on neuronal apoptosis and the expression of caspase-3,B-cell lymphoma/leukemia-2(Bcl-2),Bcl-2 associated X protein(Bax)and DDR1 were observed.Hematoxylin-eosin staining was used to observe the morphological changes of brain neurons in each group;TdT-mediated dUTP nick end labeling staining was used to detect neuronal apoptosis.Results Compared with the sham operation group,the vertebral body cells in the CA1 area of the cerebral ischemia-reperfusion group were sparse,and a large number of vacuoles,widened intercellular substanc,and nuclear pyknosis were observed,and the neuronal apoptosis rate was significantly increased[(50.25±8.50)%vs(8.98±2.50)%](P<0.01),the protein expressions of caspase-3,Bax and DDR1 in the brain tissue were significantly up-regulated[(2.58±0.38)vs(0.71±0.20),(1.82±0.43)vs(0.72±0.27),(2.33±0.58)vs(1.21±0.44)](P<0.05),Bcl-2 expression was significantly down-regulated[(0.61±0.21)vs(1.21±0.40)](P<0.05).Compared with cerebral ischemia-reperfusion group,the CA1 area of the aspirin intervention group had increased vertebral cells,reduced vacuoles,narrowede intercellular substance,and decreased neuronal cell apoptosis[(30.25±9.44)%vs(50.25±8.50)%](P<0.05),the expression of Caspase-3,Bax and DDR1 proteins in brain tissue was significantly down-regulated[(1.47±0.57)vs(2.58±0.38),(1.21±0.31)vs(1.82±0.43),(1.58±0.48)vs(2.33±0.58)](P<0.05),the expression of Bcl-2 was significantly up-regulated[(1.02±0.31)vs(0.61±0.21)](P<0.05).Conclusion Aspirin can improve nerve injury in rats with cerebral ischemia-reperfusion,and its mechanism may be related to inhibiting DDR1 protein expression and reducing apoptosis.