穴位埋线对溃疡性结肠炎大鼠肠黏膜上皮紧密连接的影响  

Effect of acupoint thread-embedding on tight junction of intestinal mucosa epithelium in rats with ulcerative colitis

作　　者：王欢 朱莹 WANG Huan;ZHU Ying(Yueyang Hospital Affiliated to Hunan University of CM,Yueyang 414000,China;First Affiliated Hospital of Hunan University of CM,Changsha 410007)

机构地区：[1]湖南中医药大学附属岳阳医院,岳阳414000 [2]湖南中医药大学第一附属医院,长沙410007

出　　处：《中国针灸》2021年第8期899-905,共7页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金面上项目:81874466;湖南省中医药管理局重点课题项目:201903。

摘　　要：目的:观察穴位埋线对"虚、瘀"状态下溃疡性结肠炎(UC)大鼠肠黏膜上皮紧密连接的影响,并探讨其作用机制。方法:60只雄性SD大鼠随机分为对照组(12只)和造模组(48只),采用腺嘌呤、番泻叶分阶段灌胃制备大鼠"虚、瘀"状态,然后用5%2,4,6-三硝基苯磺酸(TNBS,100 mg/kg)加0.25 mL 50%乙醇灌肠诱导大鼠UC模型。选取造模成功的30只大鼠随机分为模型组、药物组和埋线组,每组10只。埋线组取双侧"足三里""天枢""膈俞""脾俞""肾俞""大肠俞"穴进行埋线治疗,每14天1次,共3次;药物组采用柳氮磺吡啶肠溶片灌胃治疗,每日1次;对照组、模型组和埋线组同时予等量0.9%氯化钠溶液,均干预42 d。观察各组大鼠的一般情况及体质量变化,采用肉眼及HE染色观察大鼠结肠组织外观及形态学变化,ELISA法检测各组大鼠血清中钙调素依赖性蛋白激酶Ⅱ(CaMKⅡ)和肌球蛋白轻链激酶(MLCK)的含量,实时荧光定量PCR法检测各组大鼠结肠组织肠黏膜上皮紧密连接蛋白occludin、claudin1、claudin2、ZO-1 mRNA水平,Western blot法检测各组大鼠结肠组织肠黏膜上皮紧密连接蛋白occludin、claudin1、claudin2、ZO-1及CaM KⅡ、MLCK的蛋白水平。结果:与对照组比较,模型组大鼠体质量下降(P<0.01),结肠组织明显肿胀,出血,大量炎性细胞浸润,结肠组织学损伤指数评分升高(P<0.01),血清CaM KⅡ和MLCK含量升高(P<0.01),结肠组织occludin、claudin1和ZO-1 mRNA和蛋白水平降低(P<0.01),claudin2 mRNA和蛋白水平升高(P<0.01),CaMKⅡ和MLCK蛋白水平升高(P<0.01)。与模型组比较,药物组和埋线组大鼠体质量增加(P<0.01),结肠病变明显减轻,组织学损伤指数评分降低(P<0.01),血清CaMKⅡ和MLCK含量降低(P<0.01),结肠组织内occludin、claudin1和ZO-1 mRNA和蛋白水平增高(P<0.01,P<0.05),claudin2 mRNA和蛋白水平降低(P<0.01),CaMKⅡ和MLCK蛋白表达水平降低(P<0.01,P<0.05)。埋线组和药物组各指标比较差异无�Objective To observe the effect of acupoint thread-embedding on tight junction of intestinal mucosal epithelial barrier in rats with ulcerative colitis(UC) under the state of "deficiency and stasis", and to explore its mechanism. Methods Sixty male SD rats were randomly divided into a control group(n=12) and a UC model group(n=48). The rats were gavaged with adenine and folium sennae by stages to prepare the "deficiency and stasis" state, and then 5% 2,4,6-trinitrobenzene sulfonic acid(TNBS, 100 mg/kg) and 0.25 mL 50% ethanol were used to induce UC model. Thirty rats with successful model establishment were selected and randomly divided into a model group, a medication group and a thread-embedding group, 10 rats in each group. The rats in the thread-embedding group were intervened with thread-embedding at "Zusanli"(ST 36), "Tianshu"(ST 25), "Geshu"(BL 17), "Pishu"(BL 20), "Shenshu"(BL 23) and "Dachangshu"(BL 25), once every 14 days, totaling 3 times. The rats in the medication group were gavaged with sulfasalazine, once a day. The rats in the control group, model group and thread-embedding group were gavaged with the same amount of 0.9% sodium chloride solution at the same time for 42 days. The general condition and body weight of rats in each group were observed. The appearance and morphological changes of colonic tissue were observed by naked eye and HE staining. The serum levels of calmodulin-dependent protein kinaseⅡ(CaMKⅡ) and myosin light chain kinase(MLCK) were detected by ELISA. The mRNA levels of colonic tight junction(TJ) proteins including occludin, claudin1, claudin2 and ZO-1 were detected by quantitative real-time PCR. The protein expressions of occludin, claudin1, claudin2, ZO-1 and CaMKⅡ, MLCK were detected by Western blot. Results Compared with the control group, in the model group the body weight was decreased(P<0.01), colon tissue was significantly swelled with bleeding and a large number of inflammatory cells infiltration, and the colon tissues damage index score was increased(P<0.01), se

关 键 词：溃疡性结肠炎 穴位埋线 肠黏膜上皮 紧密连接 CaMKⅡ MLCK 

分 类 号：R245.91[医药卫生—针灸推拿学]