葛根素通过核转录因子κB信号通路抑制缺血再灌注心肌细胞损伤的机制研究

Inhibition of puerarin on myocardial ischemia reperfusion injury via nuclear facter kappa B signal pathway

目的探讨葛根素通过核转录因子κB(NF-κB)信号通路抑制缺血再灌注心肌细胞损伤的机制。方法将30只SD大鼠随机分为假手术组、模型组和预处理组,每组10只。假手术组予以冠状动脉左前降支穿线不结扎,并于穿线前给予2.5 m L·kg^(-1)0.9%氯化钠;模型组和预处理组予以结扎大鼠冠状动脉左前降支建立心肌缺血再灌注模型,并分别于结扎前经颈静脉注射2.5m L·kg^(-1)0.9%NaCl,200 mg·kg^(-1)葛根素。造模4 h后,处死大鼠并取心肌组织,用流式细胞术检测3组大鼠心肌细胞凋亡情况,用紫外分光光度法检测Caspase 3的活性,用免疫印迹法检测B淋巴细胞瘤基因-2(Bcl-2)、Bcl-2相关X蛋白(Bax)及NF-κB p65通路激活情况。结果与假手术组比较,模型组的心肌细胞凋亡率提高,Bcl-2表达量下降,Bax表达量、Caspase 3活性及NF-κB p65磷酸化水平提高(P<0.01)。与模型组比较,预处理组的心肌细胞凋亡率降低,Bcl-2表达量提高,Bax表达量、Caspase 3活性及NF-κB p65磷酸化水平降低(P<0.01)。结论葛根素预处理可抑制缺血再灌注导致的大鼠心肌细胞凋亡,可能与抑制NF-κB信号通路有关。

Objective To explore the inhibition of puerarin on myocardial ischemia reperfusion injury via nuclear factor kappa B（NF-κB） signal pathway. Methods Thirty SD rats were randomly divided into sham operation group,model group and pretreatment group,each group has 10 SD rats. For the rats in sham operation group, their left anterior descending branchs of coronary artery were not ligated and were given2. 5 m L· kg^-10. 9% NaCl. Model group and pretreatment group were used to establish myocardial ischemia reperfusion model in the left anterior descending coronary artery in rats,and rats were given 2. 5 m L·kg^-1 0. 9% NaCl or 200 mg · kg^-1puerarin by jugular vein injection before ligation of the jugular vein. After building 4 h,the rats were executed and myocardial specimens were isolated. Myocardial cell apoptosis was measured by flow cytometry. The activity of Caspase 3 was measured by spectrophotometry method. The expression of B-cell lymphoma-2（Bcl-2）,Bcl-2 associated X protein（Bax）,NF-κB p65 was detected by western blot. Results Compared with the sham operation group,myocardial cell apoptosis rate,the activity of Caspase 3,the expressionof Bax and the phosphorylation of NF-κB p65 increased,the expression of Bcl-2 decreased in model group（P〈0. 01）. Compared with model group,myocardial cell apoptosis rate,the activity of Caspase 3,the expression of Bax and the phosphorylation of NF-κB p65 decreased,the expression of Bcl-2 increased in pretreatment group（P〈0. 01）. Conclusion Puerarin pretreatment suppressed the apoptosis of myocardial cell with ischemia reperfusion,which was related with inhibition of NF-κB signaling pathway.