电针人中穴对脑梗死大鼠ACE-AngⅡ/ACE2-Ang(1-7)轴相关因子表达的干预效应研究

Effect of electro-acupuncture at Renzhong(DU26)on the expression of ACE-AngⅡ/ACE2-Ang(1-7)axis related factors in rats with cerebral infarction

[目的]观察脑梗死大鼠血管紧张素转化酶(ACE)-血管紧张素(Ang)Ⅱ/ACE2-Ang(1-7)轴通路的变化规律以及电针干预效应,从血管舒缩的角度探讨针刺治疗脑梗死的作用机制。[方法]将126只大鼠随机分为空白组、模型组、电针组。模型组和电针组按术后1、3、6、9、12、18、24 h,3、7、12 d各分为10个时相组,造模后电针组每日针刺人中穴。各组大鼠进行神经功能缺损,脑梗死面积以及免疫组化的检测。[结果]1)针刺后各组大脑中动脉闭塞(MCAO)模型大鼠神经功能缺损评分(NSS)随缺血时间延长逐渐下降,各时相评分均显著高于空白组(P<0.01);电针组各时相评分低于模型组,两组在3、7、12 d差异有统计学意义(P<0.01)。2)绿化三苯四氮唑(TTC)染色结果模型组与空白组比较差异有统计学意义(P<0.05,P<0.01),各时相电针组明显小于同时相模型组(P<0.05,P<0.01)。3)免疫组化结果:模型组ACE表达在术后上调至18 h回落,1 h~7 d显著高于空白组(P<0.01);电针组除6 h外表达均低于模型组,9、18、24 h有统计学差异(P<0.05,P<0.01)。模型组AngII表达在术后上调至24 h回落,在3~9 h和18 h~7 d高于空白组(P<0.05,P<0.01);电针组1 h~3 d均低于模型组,3、6、18 h~3 d差异有统计学意义(P<0.05,P<0.01)。模型组血管紧张素受体AT1表达在术后9 h上调至24 h回落,其中3 h、9 h~24 h有统计学差异(P<0.05,P<0.01);电针组自9 h后均低于模型组,12、18、24 h差异有统计学意义(P<0.05)。电针组ACE2表达在术后3 h上调至18 h后下调,1 h到12 d高于空白组,1、9到3 d有统计学差异(P<0.05);电针组24 h~3 d时相与模型组比较差异有统计学意义(P<0.05)。模型组Ang(1-7)表达自9 h上调至12 h后下调,除1、6 h时相外,均显著高于空白组(P<0.05,P<0.01)。电针组从3 h开始上调至24 h后下调,24 h到7 d高于模型组差异有统计学意义(P<0.01)。模型组血管内皮细胞表达的Ang(1-7)的特异性受体(MAS)表达自12 h上调至3 d后下调,3、6、12 h~12 d时相均高于空白组,12 h~7 d差异有统计学意义(P<0.05,P<0.01);电针组自9 h后表达高于模型组,其中3、24 h,3、7 d差异有统计学意义(P<0.05)。[结论]电针干预能显著改善MCAO大鼠的神经功能症状,减少脑缺血体积,改善脑血流量,下调ACE、AngII、ATl的表达情况,上调Ang(1-7)、ACE2、MAS的表达情况,改善脑梗死的预后。

[Objective]The purpose of this study is to observe the change of ACE-AngⅡ/ACE2-Ang(1-7)axis pathway in rats with cerebral infarction and the effect of electro-acupuncture(EA)intervention.The mechanism of acupuncture in the treatment of cerebral infarction was discussed from the point of vasodilation.[Methods]The 126 rats were randomly divided into blank control,model group and EA group.The model group and EA group were divided into 10 time groups according to postoperative 1,3,6,9,12,18,24 h,3,7 and 12 d.After modeling,the EA group was acupunctured at Renzhong(DU26)acupoint every day.The neurological deficit,cerebral infarction area,cerebral blood flow(CBF)and immuno-histochemistry were detected.[Results]1)After acupuncture,the NSS score of MCAO model rats decreased gradually with the prolongation of ischemia time,and the scores of each time phase were significantly higher than those in the blank control(P<0.01);the scores of the EA group were lower than those in the model group,and the differences were significant between the at 3,7 and 12 d(P<0.01).2)TTC staining results showed that compared with the blank control(P<0.05,P<0.01),and the EA group in each phase was significantly less than the model group in the same time phase(P<0.05,P<0.01).3)Immuno-histochemical results:the expression of ACE in the model group was up-regulated to 18 hours and down-regulated,which was significantly higher than that in the blank control from 1 h-7 d(P<0.01);the expression of ace in the EA group was lower than that in the model group except 6 h,with significant differences at 9 h 18 h and 24 h(P<0.05,P<0.01).The expression of AngⅡin model group was up-regulated to 24 h after operation,and significantly higher than blank control(P<0.05,P<0.01)at 3-9 h and 18 h-7 d,and the difference was significant in 3 h,6 h and 18 h-3 d(P<0.05,P<0.01).AT1 expression in model group was up-regulated to 24 h after operation,and there was significant difference in 1 h and 9 h-24 h(P<0.05,P<0.01).The expression of AT1 in EA group was lower than that in model group after 9 h,and the difference was significant in 12,18,24 h(P<0.05).The expression of ACE2 in EA group was up-regulated from 3 h to 18 h after operation,and was higher than that in the blank control at 1 h-12 d,and there was significant difference in 1 h,9 h-3 d(P<0.05),and the difference was significant in the EA group at 24 h-3 d(P<0.05).The expression of Ang(1-7)in model group was up-regulated from 9 h to 12 h after operation,except for 1 h,6 h phase,which was significantly higher than that in the blank control(P<0.05,P<0.01).The EA group was up-regulated from 3-24 h later,and was higher than that in the model group from 24 h-7 d,and the difference was significant(P<0.05).The expression of MAS in model group was up-regulated from 12 h to 3 d,and the phase of MAS in the model group was higher than that in the blank control at 3,6 h and 12 h-12 d,and the difference was significant(P<0.05,P<0.01);the expression of the EA group was higher than that in the model group after 9 h,among which the difference was significant(P<0.05).[Conclusion]EA intervention can significantly improve the neurological symptoms of MCAO rats,reduce the volume of cerebral ischemia,improve CBF perfusion,down regulate the expression of ACE,AngⅡand ATL,up regulate the expression of ANG(1-7),ACE2 and MAS,and improve the prognosis of cerebral infarction.