加味温胆汤对脑缺血再灌注损伤大鼠酪氨酸蛋白激酶2与信号转导子和转录激活子3蛋白表达及细胞凋亡的影响

Effect of Jiaweiwendan Decoction on expressions of janus activated kinase 2/signal transducer and activator of transcription 3 proteins and cell apoptosis in rats with cerebral ischemia-reperfusion injury

目的探讨加味温胆汤对脑缺血再灌注损伤(CIRI)大鼠酪氨酸蛋白激酶2(JAK2)与信号转导子和转录激活子3(STAT3)蛋白表达及细胞凋亡的影响。方法选用健康雄性SD大鼠120只,随机分为4组,每组30只。正常对照组无任何干预;假手术组仅切开颈部皮肤,暴露左侧颈总动脉;脑缺血再灌注模型组、加味温胆汤治疗组均采用线栓法制备大鼠左侧大脑中动脉脑缺血再灌注模型,然后分别给予蒸馏水灌胃和加味温胆汤灌胃。分别于缺血再灌注后3、12、24、48、72 h对各组大鼠进行神经功能Zea Longa评分并检测脑组织JAK2、STAT3蛋白表达及神经细胞凋亡情况。结果 (1)正常对照组及假手术组大鼠无神经功能缺损表现;脑缺血再灌注模型组、加味温胆汤治疗组大鼠均出现神经功能缺损,Zea Longa评分均高于正常对照组及假手术组(P<0.05)。(2)与正常对照组、假手术组大鼠相比,各时间点脑缺血再灌注模型组、温胆汤加味治疗组大鼠磷酸化JAK2、磷酸化STAT3灰度值明显降低(P<0.05),缺血再灌注后24 h温胆汤加味治疗组磷酸化JAK2、磷酸化STAT3灰度值表达高于脑缺血再灌注模型组(P<0.05)。(3)与正常对照组及假手术组大鼠比较,各时间点脑缺血再灌注模型组、加味温胆汤治疗组大鼠脑细胞凋亡率明显升高(P<0.05),而缺血再灌注后24 h加味温胆汤治疗组大鼠脑细胞凋亡率低于脑缺血再灌注模型组(P<0.05)。结论脑缺血再灌注引发JAK2、STAT3通路激活加重神经细胞损伤和凋亡,加味温胆汤可通过抑制JAK2/STAT3通路活化减少神经细胞凋亡,从而减轻CIRI。

Objective To investigate the effect of Jiaweiwendan Decoction on the expressions of janus activated kinase 2（ JAK2） /signal transducer and activator of transcription 3（ STAT3） proteins and cell apoptosis in rats with cerebral ischemia-reperfusion injury. Methods A total of 120 healthy male SD rats were randomly divided into 4 groups,with 30 rats in each group. No intervention was conducted in the normal control group. In the sham-operation group,an incision was performed in the neck skin and left common carotid artery was exposed. Rat models of left middle cerebral artery ischemia-reperfusion injury were established using suture embolization method in the cerebral ischemia-reperfusion group and Jiaweiwendan Decoction group. Intragastric administration with distilled water and Jiaweiwendan Decoction were performed in the cerebral ischemia-reperfusion group and Jiaweiwendan Decoction group respectively. At 3,12,24,48 and 72 hours after ischemia-reperfusion,Zealonga score for neurological function was assessed,the expressions of JAK2 / STAT3 proteins and nerve cell apoptosis were detected in each group. Results ① No neurological function impairment was found in the normal control group and sham-operation group. In the cerebral ischemia-reperfusion group and Jiaweiwendan Decoction group,neurological function impairment was observed,and the Zealonga scores were significantly higher compared to the normal control group and sham-operation group（ P〈0. 05）. ② At each time point,the cerebral ischemia-reperfusion group and Jiaweiwendan Decoction group obtained significantly lower gray values of JAK2 and STAT3 compared to the normal control group or sham-operation group（ P〈0. 05）,and the gray values of JAK2 and STAT3 in the Jiaweiwendan Decoction group were higher than those in the cerebral ischemia-reperfusion group（ P〈0. 05）. ③ At each time point,the cerebral ischemia-reperfusion group and Jiaweiwendan Decoction group obtained significantly higher apoptosis rate of brain cells compared to the normal control group or sham-operation group（ P〈0. 05）,and the apoptosis rate in the Jiaweiwendan Decoction group were lower than that in the cerebral ischemia-reperfusion group（ P〈0. 05）. Conclusion Cerebral ischemia-reperfusion can activate the JAK2 / STAT3 pathway and then aggravates the injury and apoptosis of brain cells. Jiaweiwendan Decoction can reduce the cell apoptosis by suppressing the activation of JAK2 / STAT3 pathway,and then alleviates the cerebral ischemia-reperfusion injury.