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标本
配穴
针灸
腹泻
型肠易
血清
炎症
因子
表达
影响
李奕宏
5【论著】标本配穴针灸法对腹泻型肠易激综合征模型大鼠紧密连接及血清炎症因子表达的影响李奕宏,胡江杉,李佳,王华(湖北中医药大学/针灸治未病湖北省协同创新中心,湖北 武汉 430061)摘要:目的观察标本配穴针灸对腹泻型肠易激综合征模型大鼠一般体征、胃排空率、小肠推进率、肠组织形态学及血清中白介素-6(IL-6)、白介素-8(IL-8)及肿瘤坏死因子-(TNF-)表达的影响。方法选取40只SPF级雌性Wistar大鼠随机分为空白组、模型组、标本配穴组、常规配穴组。以急慢性应激相结合CAS方法制备腹泻型肠易激综合征大鼠模型。造模同时测定大鼠粪便含水量、胃排空率、小肠推进率、透射电镜观察大鼠肠黏膜超微结构、Elisa法检测大鼠血浆中IL-6、IL-8和TNF-的含量。结果与空白组相比,其他3组的粪便含水量升高(P0.05);标本配穴组和常规配穴组较模型组粪便含水量降低(P0.05),且标本配穴组低于常规配穴组(P0.05)。与空白组比较,其他3组胃排空率均降低,小肠推进率均增高(0.05);标本配穴组与常规配穴组较模型组大鼠胃排空率增高,小肠推进率降低(P0.05);与空白组相比,模型组大鼠肠黏膜结构损伤,治疗后标本配穴组和常规配穴组较模型组大鼠的肠黏膜超微结构相对完整,且标本配穴组优于常规配穴组。与空白组相比,其他3组血清中IL-6、IL-8和TNF-含量均增高(P0.05),标本配穴组和常规配穴组较模型组血清中 IL-6、IL-8和 TNF-含量均降低(P0.05),且标本配穴组低于常规配穴组(P0.05)。结论标本配穴针灸可以改善腹泻型肠易激大鼠一般行为学体征、胃排空率、小肠推进率及肠组织形态学,降低血清炎症因子IL-6、IL-8和TNF-的表达,从而改善腹泻型肠易激综合征患者的症状。关键词:针灸;标本配穴;肠易激综合征;白介素-6;白介素-8;肿瘤坏死因子-中图分类号:R285.5文献标识码:Adoi:10.3969/j.issn.1008 987x.2023.02.01Effect of acupuncture andmoxibustion with“Biao-Ben”acupoint combinationon ti-ght junction and inflammatory factor in serum in IBS-D model ratsLI Yi-hong,HU Jiang-shan,LI Jia,WANG Hua(Hubei University of Chinese Medicine/Hubei Collaborative Innovation Center of Acupuncture and Moxibustion forDisease Prevention,Wuhan 430061)Abstracts:ObjectiveTo observe the general signs,gastric emptying rate,small intestine propulsion rate,intestinal his-tomorphologyandserum content of diarrhea-predominant irritable bowel syndrome modelrats with acupuncture and acupunctureEffects of interluekin-6(IL-6),IL-8 and tumor necrosis factor-(TNF-)expression.MethodsForty SPF female Wistar ratswere randomlydivided intoblankgroup,modelgroup,“Biao-Ben”acupoint combination group,and regularacupointcombinationgroup.The irritable bowel syndrome rat model was established by combining acute and chronic stress with CAS method.UseSimultaneous determination to detect fecal water content,gastric emptying rate,small intestinal propulsion rate,transmissionelectron microscope to observe rat intestinal mucosal ultrastructure,and Elisa method to detect the contents of IL-6,IL-8 and基金项目:国家自然科学基金面上项目(项目编号:81973931);国家中医药管理局岐黄工程项目(国中医药人教函 2018 284 号)。作者简介:李奕宏(1998-),男,湖北中医药大学针灸骨伤学院 2020 级针灸推拿学专业硕士研究生,研究方向:针灸效应的机制研究。通信作者:王华(1955-),男,湖北中医药大学教授,研究方向:针灸效应的机制研究,E-mail:。6 TNF-in ratplasma.ResultsComparedwith blankgroup,the fecal water contentofthe other three groups increased(P0.05);the fecal water content of“Biao-Ben”acupoint combination group and regular acupoint combination group was lower thanthat of model group(P0.05),and“Biao-Ben”acupoint combination group was lower than model group in regular acupointcombinations group(P0.05);compared with blank group,the gastric emptying rate of the other three groups was increased,and the small intestine propulsion rate was decreased(P0.05);compared with blank group,the intestinal mucosal structure ofthe rats inmodelgroupwasdamaged,andthe ultrastructure of the intestinalmucosa ofthe ratsin“Biao-Ben”acupointcombinationgroup and regular acupoint combination group after treatment was relatively more intact compared with model group,and“Biao-Ben”acupoint combination group was better than regular acupoint combinations group;compared with blank group,the serum levels of IL-6,IL-8 and TNF-in the other three groups were all increased(P0.05);compared with model group,the levels of IL-6,IL-8 and TNF-of“Biao-Ben”acupoint combination group and regular acupoint combination group weredecreased(P0.05),and the“Biao-Ben”acupoint combination group was lower thanregular acupoint combination group(P0.05).ConclusionAcupuncture with acupoints of“Biao-Ben”acupoint combination can improve the general behavioralsigns,gastric emptying rate,small intestinal propulsion rate and intestinal histomorphology in IBS-D rats,and reduce theexpressions of serum inflammatory factors IL-6,IL-8 and TNF-,thereby reducing the inflammatory factors and relievingsymptoms in patients with diarrhea-predominant irritable bowel syndrome.Keywords:Acupuncture;“Biao-Ben”acupoint combination;Irritable bowel syndrome;IL-6;IL-8;TNF-肠易激综合征(Irritablebowelsyndrome,IBS)是一种以肠道功能紊乱为主要表现的疾病,其临床症状主要表现为持续存在且间断发作的腹痛、腹部不适、以及排便异常。还可出现睡眠障碍、精神抑郁或焦躁、慢性疲劳综合征等胃肠道外症状1。现代医学根据罗马诊断标准将 IBS 分为4种类型,分别为腹泻型(diarrhea-predominantIBS,IBS-D)、便秘型(constipation-predomi-nantIBS,IBS-C)、混合型(mixed-typeIBS,IBS-M)和不定型(unsubtypedIBS,IBS-U),且以 IBS-D 发病率最高2-4。目前临床研究证实针灸可以明显改善 IBS-D患者的腹痛和腹泻症状5-6,也有实验证实针灸能调控多个炎性因子7,而且配穴及组方多样8。同时有研究表明 IBS 患者存在肠上皮紧密连接破坏9,出现腹痛、腹泻、大便性状改变等一系列症状。本研究通过观察标本配穴针灸对 IBS 模型大鼠血清中白介素-6(IL-6)、白介素-8(IL-8)、肿瘤坏死因子-(TNF-)含量,肠组织形态学及胃肠转运功能的调控,探讨标本配穴针灸在改善IBS-D方面的效应,为进一步探究针灸治疗IBS-D的科学机制提供客观依据。1材料与方法1.1实验动物3月龄 SPF 级雌性 Wistar 健康大鼠40只,体质量(22020)g,购买于湖北省动物实验研究中心,动物许可证号:SCXK(鄂)2020-0018,饲养于湖北中医药大学针灸研究所,实验室温度维持于2225,昼夜节律交替的时间为12h/12h,进行1周适应性喂养后行分组、造模、治疗。1.2主要试剂与仪器大鼠 IL-6ELISA 试剂盒、大鼠 IL-8ELISA 试剂盒、大鼠 TNF-ELISA 试剂盒(均购于赛默飞世尔科技(中国)有限公司,批号:324017-002,324317-002,291276-001)。ST-360型酶标仪(上海旦鼎国际贸易有限公司);aquaplus2系列艾科浦超纯水制成机(美国艾科浦国际有限公司);TGL-10B 系列台式离心机(上海安亭科学仪器厂)。1.3实验分组及造模干预大鼠适应性喂养7d 后开始进行实验,采用随机数字表法,将40只大鼠随机分为4组,分别为空白组、模型组、标本配穴组和常规配穴组,每组各10只。空白组:正常喂养,不做任何干预;将其余各组大鼠以急慢性应激相结合 CAS 方法制备 I