加味益气通玄方对大鼠脑缺血再灌注损伤的抑制作用

目的探究加味益气通玄方对大鼠脑缺血再灌注损伤的抑制作用,为临床用药提供科学依据。方法72只大鼠随机分为6组,即假手术组、模型组、阿司匹林组、加味益气通玄方(27.68、13.84、6.92 g/kg)组,每组12只。除假手术组外,其余各组大鼠通过中动脉栓塞法(MCAO)缺血2 h后再灌注,再灌注24 h后分别给予27.68、13.84、6.92 g/kg加味益气通玄方或60 mg/kg阿司匹林治疗,1次/d,持续14 d。修正版神经功能缺损评分(mNSS)、氯化三苯基四氮唑(TTC)染色、苏木精-伊红(HE)染色、原位末端转移酶标记(TUNEL)评估加味益气通玄方对大鼠CRI的保护作用。蛋白免疫印迹实验检测PI3K/Akt-eNOS信号通路相关蛋白,以及凋亡相关蛋白Bcl-2、Bax、cleaved-Caspase3的表达。结果加味益气通玄方能明显降低大鼠的神经行为学评分,减少梗塞率,减轻脑组织病理学损伤,降低脑组织细胞凋亡;增加PI3K/Akt-eNOS信号通路相关蛋白p-Akt、p-NOS的表达,提高抗凋亡蛋白Bcl-2的表达,降低促凋亡蛋白Bax、cleaved-Caspase3的表达以及Bax/Bcl-2比值。结论加味益气通玄方可以改善大鼠脑缺血再灌注损伤,其作用机制可能与激活PI3K/Akt-eNOS通路有关。

Effect of Supplemented Yiqi Tongxuan Formula on inhibiting cerebral ischemia reperfusion injury in rats

AIM To investigate the mechanism of supplemented Yiqi Tongxuan Formula on cerebral ischemia reperfusion injury in rats to clarify the scientific basis for its clinical medication.METHODS Seventy-two rats were randomly divided into 6 groups,namely the sham group,the model group,aspirin group,and groups of supplemented Yiqi Tongxuan Formula(27.68 g/kg,13.84 g/kg,6.92 g/kg),with 12 rats in each group.Except for the sham group,the rats in the other groups were subjected to middle cerebral artery occlusion(MCAO)ischemia for 2 h,and then the embolus removal for reperfusion.And 24 h later,the rats went on corresponding 14 d course intervention of 27.68 g/kg/d,13.84 g/kg/d,6.92 g/kg/d of supplemented Yiqi Tongxuan Formula,and 60 mg/kg/d of aspirin.Modified neurological severity scores(mNSS),triphenyltetrazol chloride(TTC)staining,hematoxylin-eosin(HE)staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL)were used to assess the protective effect of supplemented Yiqi Tongxuan Formula on cerebral ischemia reperfusion injury rats.Western blot was used to detect the expression of PI3K/Akt-eNOS signaling pathway-related proteins and apoptosis-related proteins Bcl-2,Bax,and cleaved-Caspase3.RESULTS Supplemented Yiqi Tongxuan Formula significantly reduced the neurobehavioral score,the infarct rate,the pathological damage of brain tissue,and the apoptotic rate of brain tissue.It also increased the PI3K/Akt-eNOS signaling pathway-related p-Akt and p-eNOS protein expression,increased the expression of anti-apoptotic protein Bcl-2,decreased the expression of pro-apoptotic proteins Bax,cleaved-Caspase3 and the ratio of Bax/Bcl-2.CONCLUSION Supplemented Yiqi Tongxuan Formula can improve cerebral ischemia reperfusion injury in rats,and its mechanism may be associated with the activation of PI3K/Akt-eNOS pathway.