电针足三里穴对肠缺血再灌注损伤大鼠小肠黏膜上皮紧密连接蛋白ZO-1的调节作用  

Regulatory effect of electroacupuncture at Zusanli point on small intestinal mucosal epithelial tight junction protein ZO-1 in rats with intestinal ischemia-reperfusion injury

作　　者：杜明华[1,2] 郭润[3] 张文华 胡森[2] Du Minghua;Guo Run;Zhang Wenhua;Hu Sen(Department of Emergency,Medical Innovation Research Division,First Medical Center of Chinese PLA General Hospital,Beijing 100853,China;Research Center of Trauma Repair and Tissue Regeneration,Medical Innovation Research Division,First Medical Center of Chinese PLA General Hospital,Beijing 100853,China;Department of Dermatology,Guang′anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China)

机构地区：[1]解放军总医院第一医学中心急诊医学科,北京100853 [2]解放军总医院第一医学中心医学创新研究部创伤修复与组织再生研究中心,北京100853 [3]中国中医科学院广安门医院皮肤科,北京100053

出　　处：《中华损伤与修复杂志（电子版）》2022年第4期315-321,共7页Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)

基　　金：国家自然科学基金面上项目(81774422);解放军总医院苗圃项目青年基金(18KMM40)。

摘　　要：目的探讨电针足三里穴对肠缺血再灌注损伤大鼠小肠黏膜上皮紧密连接蛋白闭锁小带蛋白(ZO)-1的调节作用。方法按随机数字表法将30只SD大鼠分为模型组、电针足三里穴组、电针非经非穴组,每组10只。每组大鼠均于麻醉后沿腹正中线切口,分离肠系膜上动脉,用无损伤血管夹夹闭其起始部位,30 min后松夹恢复血流60 min,造成小肠缺血/再灌注损伤。电针足三里穴组于大鼠缺血后即刻行电针双侧足三里穴30 min,强度为2~3 mA,频率2~100 Hz;电针非经非穴组采用相同频率和强度于缺血后即刻刺激大鼠非经非穴(足三里穴外侧旁开0.5 cm)30 min;模型组不做任何治疗。于缺血再灌注损伤60 min时,以腹主动脉放血法处死各组大鼠,留取每只大鼠的血液和10 cm远端回肠组织。将留取的大鼠血液经10000×g,4℃离心10 min后取上清得到血浆20μL,全自动生化分析仪检测大鼠血浆脏器功能指标,包括谷丙转氨酶(GPT)、肌酸激酶同工酶(CK-MB)、尿素氮、肌酐;免疫荧光染色及蛋白质印迹法检测各组大鼠小肠黏膜上皮紧密连接蛋白ZO-1分布情况与表达水平。数据比较采用单因素方差分析和LSD-t检验。结果缺血再灌注损伤60 min时,模型组大鼠GPT、CK-MB、尿素氮和肌酐水平分别为(88.1±11.6)μ/L、(482.3±69.7)μ/L、(11.6±3.7)mmol/L、(52.3±13.2)μmol/L,电针足三里穴组大鼠分别为(37.3±6.4)μ/L、(213.7±44.3)μ/L、(5.2±1.4)mmol/L、(30.1±5.7)μmol/L,电针非经非穴组大鼠分别为(85.7±13.4)μ/L、(460.2±72.3)μ/L、(10.8±4.2)mmol/L、(53.4±12.1)μmol/L,3组大鼠各指标比较,差异均有统计学意义(F=69.4063、55.3611、10.9582、14.6817,P<0.05)。与模型组比较,电针足三里穴组大鼠血浆中GPT、CK-MB、尿素氮和肌酐水平与均降低,2组比较差异均有统计学意义(t=12.126、10.285、5.116、4.883,P<0.05),而电针非经非穴组大鼠血浆中GPT、CK-MB、尿素氮和肌酐水平与模型组相近,Objective To investigate the regulative effects of electroacupuncture at Zusanli point on the tight junction protein zonula occludens(ZO)-1 of small intestinal mucosal epithelium in rats with intestinal ischemia-reperfusion injury.Methods Thirty SD rats were divided into model group,electroacupuncture Zusanli group,electroacupuncture non-meridian non-acupoint group according to the random number table method,10 rats in each group.After anesthesia,the rats in each group were cut along the median abdominal line,the superior mesenteric artery was separated,and its initial position was clamped with a non-invasive vascular clamp.After 30 minutes,the clamp was released to restore blood flow for 60 minutes,causing intestinal ischemia/reperfusion injury.In the electroacupuncture Zusanli group,electroacupuncture was performed at Zusanli point for 30 minutes immediately after ischemia,with an intensity of 2-3 mA and a frequency of 2-100 Hz;in the electroacupuncture non-meridian non-acupoint group,rats were stimulated at non-meridian and non-acupoints with the same frequency and intensity immediately after ischemia(0.5 cm beside the lateral side of Zusanli point)for 30 minutes;the model group received no treatment.At 60 minutes of ischemia-reperfusion injury,the rats in each group were killed by abdominal aortic bloodletting,and the blood and 10 cm distal ileal tissue of each rat were retained.The collected rat blood was centrifuged at 10000×g for 10 min at 4℃and the supernatant was taken to obtain 20μL of plasma.An automatic biochemical analyzer was used to detect the functional indexes of the rat plasma organs,including glutamic-pyruvic transaminase(GPT),creatine kinase isoenzyme(CK-MB),urea nitrogen and creatinine.Immunofluorescence staining and Western blotting were used to detect the distribution and expression of tight junction protein ZO-1 in the small intestinal mucosa of rats in each group.Data were compared by one-way ANOVA and LSD-t test.Results At 60 minutes of ischemia-reperfusion injury,the levels of GPT,C

关 键 词：电针 大鼠 小肠 缺血再灌注 紧密连接蛋白 足三里穴 

分 类 号：R245[医药卫生—针灸推拿学]