基于AMPK/PFK-2对经皮穴位电刺激“内关”穴防护模拟失重大鼠心肌损伤的效应机制研究  

Mechanism of TEAS at PC6 in Protecting Myocardial Injury in Rats with Simulated Microgravity Based on AMPK/PFK-2 Signaling Pathway

作　　者：张玲[1] 嵇波[1] 刘翼天 丁柏[2] 李莹辉[2] 谢亚娜 汪德生[2] 葛云鹏 方洋 李获朋 ZHANG Ling;JI Bo;LIU Yitian;DING Bai;LI Yinghui;XIE Yana;WANG Desheng;GE Yunpeng;FANG Yang;LI Huopeng(Beijing University of Chinese Medicine,Beijing 100029,China;Astronaut Center of China,Beijing 100094,China;China Agricultural University,Beijing 100193,China)

机构地区：[1]北京中医药大学,北京100029 [2]中国航天员科研训练中心,北京100094 [3]中国农业大学,北京100193

出　　处：《针灸临床杂志》2024年第6期70-77,共8页Journal of Clinical Acupuncture and Moxibustion

基　　金：国家自然科学基金面上项目,编号:81373558;航天医学基础与应用国家重点实验室开放基金,编号:SMFA16K02。

摘　　要：目的:观察经皮穴位电刺激(TEAS)对模拟失重大鼠心功能、心肌病理组织形态、心肌能量代谢和糖酵解相关因子腺苷酸活化蛋白激酶α1(AMPKα1)、磷酸果糖激酶2(PFK-2)蛋白和mRNA表达的影响,探讨TEAS防治模拟失重诱发大鼠心肌损伤的效应和机制。方法:将36只SPF级Wistar大鼠随机分为空白组、模型组和经皮电组,每组各12只;采用尾部悬吊法制备模拟失重大鼠模型,持续30 d。于造模后第2天对经皮电组选用双侧“内关”穴进行干预,疏密波,频率2 Hz/100 Hz,强度1 mA,1次/2 d,每次30 min,共14次。每周记录大鼠体质量变化,4周后测量大鼠心脏、右肢比目鱼肌质量和胫骨长度;采用小动物超声心动图检测大鼠心脏功能;HE染色观察心肌组织病理形态改变;比色法和化学发光法分别检测心肌组织ATP和AMP含量;Western blot和Real-Time PCR法检测心肌组织中AMPKα1、PFK-2蛋白和mRNA表达水平。结果:与空白组比较,模型组大鼠体质量、比目鱼肌湿重及与体质量的比值、左心室质量和心胫比均显著降低,差异具有统计学意义(P<0.01),心脏功能下降,心肌纤维排列稀疏、断裂和溶解,心肌组织ATP含量显著降低,差异具有统计学意义(P<0.01)和AMP含量升高(P>0.05),心肌组织中AMPKα1、PFK-2蛋白和mRNA表达显著降低,差异具有统计学意义(P<0.01)。与模型组比较,经皮电组体质量增加,左心室质量、心胫比显著升高,差异具有统计学意义(P<0.05),心脏功能上升,心肌纤维肌排列整齐,无明显坏死、溶解,心肌组织ATP含量显著升高和AMP显著降低,差异具有统计学意义(P<0.05),心肌组织中AMPKα1、PFK-2蛋白和mRNA表达水平显著升高,差异具有统计学意义(P<0.01或P<0.05)。结论:经皮穴位电刺激“内关”穴可改善模拟失重大鼠心脏功能和结构损伤,提高心脏能量代谢水平,其机制可能与上调心肌组织AMPKα1、PFK-2表达水平进而增强心肌糖酵解过程来防Objective:To observe the impacts of transcutaneous electrical acupoint stimulation(TEAS) on protein and mRNA expressions of cardiac function,myocardial pathological morphology,myocardial energy metabolism and glycolytic related factors adenylate activated protein kinase α1(AMPKα1) and 6-phosphofructo-2-kinase(PFK-2) in rats with simulated microgravity,thus to explore the protective effect and mechanism of TEAS on myocardium injury induced by simulated microgravity.Methods:A total of 36 SPF Wistar rats were randomly divided into the blank group,the model group,and the TEAS group,with 12 rats in each group.The simulated microgravity model was prepared by tail suspension method for 30 days.On the second day after modeling,the TEAS group was interfered with electro-needling bilateral Neiguan(PC6),density wave,frequency of 2 Hz/100 Hz,intensity of 1 mA,once every other day,30 min each time,a total of 14 times.Changes in body weight were recorded weekly;mass of heart and right soleus muscle and tibia length were measured after 4 weeks of treatment.Echocardiography was used to detect heart function,HE staining was used to observe the pathological changes of myocardial tissues,Colorimetry and chemiluminescence were used to detect the contents of ATP and AMP in myocardial tissues,and Western blot and Real-Time PCR were used to detect the protein and mRNA expressions of AMPKα1 and PFK-2.Results:Compared with those in the blank group,the body weight,wet weight of soleus muscle,and its ratio to body weight,the left ventricular mass,the ratio of heart mass to tibial length were significantly reduced in the model group(P<0.01);the cardiac function declined,and the myocardial fibers arranged sparsely,fractured,and dissolved in the model group;the content of ATP in myocardial tissues significantly decreased(P<0.01),the content of AMP significantly increased(P<0.05),and the protein and mRNA expressions of AMPKα1 and PFK-2 significantly decreased in the model group(P<0.01).Compared with those in the model group,the body weight

关 键 词：模拟失重 经皮穴位电刺激 内关穴 糖酵解 腺苷酸活化蛋白激酶 磷酸果糖激酶2 

分 类 号：R246.1[医药卫生—针灸推拿学]