[1]何妙嬴,张燚格,柳冬冬,等.电针内关预处理对心肌缺血再灌注损伤大鼠心肌PI3K/Akt通路的影响[J].陕西中医,2023,(3):298-303.[doi:DOI:10.3969/j.issn.1000-7369.2023.03.006]
　HE Miaoying,ZHANG Yige,LIU Dongdong,et al.Effect of electroacupuncture Neiguan pretreatment on myocardial PI3K/Akt pathway in rats with myocardial ischemia reperfusion injury[J].,2023,(3):298-303.[doi:DOI:10.3969/j.issn.1000-7369.2023.03.006]
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电针内关预处理对心肌缺血再灌注损伤大鼠心肌PI3K/Akt通路的影响
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2023年3期
页码:
298-303
栏目:
基础研究
出版日期:
2023-03-05

文章信息/Info

Title:
Effect of electroacupuncture Neiguan pretreatment on myocardial PI3K/Akt pathway in rats with myocardial ischemia reperfusion injury
作者:
何妙嬴; 张燚格; 柳冬冬; 李 丹; 张程程; 王 超
(湖南中医药大学,湖南 长沙 410208)
Author(s):
HE Miaoying; ZHANG Yige; LIU Dongdong; LI Dan; ZHANG Chengcheng; WANG Chao
(Hunan University of Chinese Medicine,Changsha 410208,China)
关键词:
心肌缺血再灌注;  内关;  电针预处理;  自噬;  磷脂酰肌醇-3-激酶/蛋白激酶B通路;  大鼠
Keywords:
Myocardial ischemia reperfusion;  Neiguan;  Electroacupuncture preconditioning;  Autophagy;  PI3K/Akt pathway;  Rat
分类号:
R 542.2
DOI:
DOI:10.3969/j.issn.1000-7369.2023.03.006
文献标志码:
A
摘要:
目的:观察电针内关穴预处理对心肌缺血再灌注损伤(MIRI)模型大鼠心肌组织自噬相关通路磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)的影响,探讨电针对MIRI的治疗作用机制。方法:48只SD大鼠随机分为空白组、假手术组、模型组、针刺组,每组12只。采用冠状动脉左前降支结扎法建立MIRI模型。针刺组电针双侧“内关”穴,30 min/次,1 次/d,连续干预7 d。记录不同组大鼠在造模前、结扎后40 min、松解后1 min、再灌注60 min的心电图并分析其ST段电位变化。HE染色观察心肌明显损伤部位病理变化; 免疫组化法观察大鼠心肌组织PI3K、Akt表达。Western blotting法检测心肌组织PI3K、Akt、磷酸化Akt(p-Akt)表达水平,并计算各组p-Akt/Akt比值。结果:模型组和针刺组结扎后40 min与松解后1 min的ST段电位均较假手术组升高,差异有统计学意义(均P<0.05)。模型组和针刺组松解后1 min、再灌注60 min时心电图ST段电位均较结扎后40 min降低,差异有统计学意义(均P<0.05)。针刺组再灌注60 min的心电图ST段电位较模型组低,差异无统计学意义(P>0.05)。空白组心肌组织肌纤维呈红染,排列紧密,结构完整; 假手术组心肌纤维呈规则排列,细胞核呈现蓝色,形态无异常,无明显病理变化。模型组心肌纤维断裂、紊乱,肌间隙明显增宽,间质可见出血,心肌纤维间散在炎性细胞浸润。针刺组无明显充血、水肿及炎性细胞浸润,可见少量心肌纤维断裂以及肌间隙增宽,但较模型组有较大程度减轻。从免疫组化结果来看,与空白组、假手术组相比较,模型组心肌组织中PI3K、Akt阳性表达较高,差异有统计学意义(均P<0.01)。针刺组心肌组织中PI3K、Akt阳性表达低于模型组,差异有统计学意义(均P<0.01)。从Western blotting检测结果来看,各组大鼠的Akt蛋白水平比较,差异无统计学意义(均P>0.05)。模型组PI3K、p-Akt表达水平和p-Akt/Akt比值均低于空白组和假手术组,差异有统计学意义(均P<0.05)。针刺组PI3K、p-Akt表达水平和p-Akt/Akt比值均高于模型组,差异有统计学意义(均P<0.05)。结论:电针内关预处理可通过激活MIRI大鼠心肌细胞PI3K/Akt通路抑制过度自噬,从而发挥对MIRI的治疗作用。
Abstract:
Objective:To observe effect of electroacupuncture at Neiguan on the autophagy-related pathway phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt)in myocardial tissue of rats with myocardial ischemia reperfusion injury(MIRI),and to explore the therapeutic mechanism of electroacupuncture on MIRI.Methods:48 SD rats were randomly divided into blank group,sham group,model group and acupuncture group,with 12 rats in each group.The MIRI model were established by ligating the anterior descending branch of the left coronary artery,and the acupuncture group were subjected to electric acupuncture for 30 min/time,once a day for seven days.ECG of rats in different groups were recorded before model establishment,40 min after ligation,1 min after release and 60 min after reperfusion,and the changes of ST segment potential were analyzed.HE staining was used to observe the pathological changes of the obvious injured parts of myocardium.Immunohistochemistry was performed to observe the expression of PI3K and Akt in myocardial tissues of rats.Western blotting was used to detect the expression levels of PI3K,Akt and phosphorylated Akt(p-Akt)in myocardial tissues and the p-Akt/Akt ratio calculated for each group.Results:The ST segment potentials of the model group and the acupuncture group were significantly higher than those in the sham group at 40 min after ligation and 1min after release,difference statistically significant(all P<0.05).The ST segment potentials of ECG in the model group and the acupuncture group at 1 min after release and 60 min after reperfusion were significantly lower than those at 40 min after ligation,difference statistically significant(all P<0.05).The ST segment potentials of ECG in the acupuncture group after 60 min reperfusion were lower than those in the model group,difference not statistically significant(P>0.05).In the blank group the myocardial fibers were red stained,tightly arranged and structurally intact.In the sham group the myocardial fibers were arranged regularly nuclei were blue morphology were normal,and there were no obvious pathological changes.In the model group myocardial fibers were broken and disordered muscle spaces were widened,interstitial hemorrhage was seen,and inflammatory cells were scattered among myocardial fibers.There was no obvious hyperemia,edema and inflammatory cell infiltration in the acupuncture group and a small amount of myocardial fiber breakage and muscle space widening were observed,but they were significantly reduced compared with the model group.From the results of immunohistochemistry,compared with the blank group and the sham group,the positive expressions of PI3K and Akt in the model group were higher,difference statistically significant(all P<0.01).Compared with the model group the positive expression of PI3K and Akt in myocardial tissue of the acupuncture group decreased,difference statistically significant(all P<0.01).There was no significant difference in Akt protein levels among the groups(all P>0.05).Compared with the blank group and the sham group,expression levels of PI3K,p-Akt and the ratio of p-Akt/Akt in the model group decreased,difference statistically significant(all P<0.05).Compared with the model group,expression levels of PI3K,p-Akt and the ratio of p-Akt/Akt in the acupuncture group increased,difference statistically significant(all P<0.05).Conclusion:Electroacupuncture Neiguan pretreatment may inhibit excessive autophagy by activating PI3K/Akt pathway in cardiomyocytes of MIRI rats,thus exerting therapeutic effect on MIRI.

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备注/Memo

备注/Memo:
基金项目:湖南省自然科学基金资助面上项目(2021JJ30507)
更新日期/Last Update: 2023-03-13