电针足三里对脓毒症大鼠肠上皮细胞间紧密连接结构的影响  

Effect of Electroacupuncture at Zusanli Point on Tight Junctions of Enterocytes in Rats with Sepsis

作　　者：楼屹[1] 朱之青 谢莉莉 冯杨荣[1] LOU Yi;ZHU Zhiqing;XIE Lili;FENG Yangrong

机构地区：[1]浙江中医药大学附属宁波市中医院,浙江宁波315010

出　　处：《新中医》2022年第15期163-168,共6页New Chinese Medicine

基　　金：宁波市自然科学基金项目(2018A610424);浙江省中医药科技计划项目(2016ZA21619)。

摘　　要：目的:研究电针足三里对脓毒症大鼠肠上皮细胞间紧密连接结构的影响及可能机制。方法:将60只SD大鼠按随机数字表法分为假手术组、模型组、电针组各20只,假手术组仅进行开关腹手术,模型组及电针组用盲肠结扎穿孔法(CLP)造模,电针组造模后电针双侧足三里,连续3 d。观察大鼠一般情况及造模后3 d生存率,于造模后3 d各组取8只存活大鼠,留取肠组织电镜观察紧密连接超微结构,免疫蛋白印迹(Western-blot)、逆转录聚合酶链式反应(RT-PCR)检测闭合蛋白-3(Claudin-3)、闭锁小带蛋白-1(ZO-1)、咬合蛋白(Occludin)的蛋白表达及mRNA表达,酶联免疫吸附法(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)浓度,并抽血测定FITC标记葡聚糖40(FD-40)浓度和D-乳酸水平。结果:造模后3 d模型组存活率60%,电针组存活率90%,较模型组明显升高(P<0.05)。与假手术组比较,模型组血清FD-40浓度、D-乳酸水平及肠组织TNF-α、IL-6浓度升高(P<0.05),肠组织Claudin-3、ZO-1、Occludin蛋白及mRNA表达均降低(P<0.05);与模型组比较,电针组FD-40浓度、D-乳酸水平及肠组织TNF-α、IL-6浓度均降低(P<0.05),肠组织Claudin-3、ZO-1、Occludin蛋白及mRNA表达均升高(P<0.05)。结论:电针足三里能明显改善脓毒症导致的紧密连接结构病理损伤,上调紧密连接蛋白及mRNA的表达,从而降低肠道通透性,其机制可能与抑制肠道炎性因子有关。Objective:To study the effect and the possible mechanism of electroacupuncture at Zusanli point(ST 36)on tight junctions of enterocytes in rats with sepsis.Methods:A total of 60 SD rats were divided into the sham operation group,the model group and the electroacupuncture group according to the random number table method,with 20 rats in each group.The sham operation group was simply given abdominal opening and closing operation,and the model group and the electroacupuncture group were modeled by cecal ligation and puncture(CLP)method.After model establishment,the electroacupuncture group was given electroacupuncture at Zusanli point on both sides for three consecutive days.The general situation of rats and the survival rate at the third day after model establishment were observed.Three days after model establishment,eight surviving rats from each group were selected;the intestinal tissues were taken to observe ultrastructure of tight junction by electroscope;the protein and mRNA expressions of claudin-3,zona occludens-1(ZO-1)and occludin were detected by western-blot and reverse transcription-polymerase chain reaction(RT-PCR);the concentration of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were detected by enzyme-linked immunosorbent assay(ELISA)method;the concentration of FITC-labeled dextran 40(FD-40)and the levels of D-lactate were measured by blood drawing.Results:Three days after model establishment,the survival rate was 60%in the model group and 90%in the electroacupuncture group,and the one in the electroacupuncture group was significantly increased when compared with that in the model group(P<0.05).The concentration of FD-40 in serum and TNF-αand IL-6 in intestinal tissues,as well as the D-lactate level in the model group were increased when compared with those in the sham operation group(P<0.05),and the protein and mRNA expressions of claudin-3,ZO-1 and occludin in intestinal tissues were decreased(P<0.05).The concentration of FD-40 in serum and TNF-αand IL-6 in intestinal tissues,as well as

关 键 词：脓毒症 电针 足三里 肠道通透性 肠上皮细胞 紧密连接结构 动物实验 大鼠 

分 类 号：R245.97[医药卫生—针灸推拿学]