基于肥大细胞脱颗粒探讨艾灸对腹泻型肠易激综合征大鼠结肠低度炎性反应的影响  

Effect of moxibustion on colonic low-grade inflammatory response in rats with diarrheapredominant irritable bowel syndrome based on mast cell degranulation

作　　者：方明 储浩然 宋小鸽[5] 阮静茹 邹玲 李奎武 廖路敏 马文黎 韩啸宇 朱敬伟 王子叶 方誉澄 FANG Ming;CHU Haoran;SONG Xiaoge;RUAN Jingru;ZOU Ling;LI Kuiwu;LIAO Lumin;MA Wenli;HAN Xiaoyu;ZHU Jingwei;WANG Ziye;FANG Yucheng(Second Clinical Medical College,Anhui University of CM,Hefei 230061,China;Famous Doctors Hall,Second Affiliated Hospital of Anhui University of CM/Anhui Acupuncture-Moxibustion Hospital,Hefei 230061;Acupuncture-Moxibustion Clinical Research Institute,Anhui Academy of TCM,Hefei 230061;Anhui Acupuncture-Moxibustion Clinical Medicine Research Center,Hefei 230061;Institute of Acupuncture and Meridians,Anhui University of CM)

机构地区：[1]安徽中医药大学第二临床医学院,合肥230061 [2]安徽中医药大学第二附属医院/安徽省针灸医院名医堂,合肥230061 [3]安徽省中医药科学院针灸临床研究所,合肥230061 [4]安徽省针灸临床医学研究中心,合肥230061 [5]安徽中医药大学针灸经络研究所

出　　处：《中国针灸》2024年第3期283-294,共12页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金面上项目:81774399;储浩然全国名老中医药专家传承工作室项目:国中医药人教函[2022] 75号;安徽省中医药领军人才项目:中医药发展秘[2018]23号。

摘　　要：目的:观察艾灸对腹泻型肠易激综合征(IBS-D)模型大鼠结肠肥大细胞脱颗粒及炎性因子表达的影响,探讨艾灸治疗IBS-D的可能机制。方法:从5只健康SPF级SD孕鼠产下的45只幼鼠中随机选取8只作为正常组,其余37只采用母子分离+醋酸灌肠+慢性束缚应激的方法制备IBS-D模型,将造模成功的32只大鼠随机分为模型组、酮替芬组、艾灸组、灸药组,每组8只。酮替芬组给予酮替芬溶液(10 mL/kg)灌胃,艾灸组给予悬灸双侧“天枢”“上巨虚”,灸药组给予悬灸双侧“天枢”“上巨虚”结合酮替芬溶液灌胃,均每天1次,连续干预7 d。分别于造模前后及干预后计算各组稀便率、检测腹壁回缩反射(AWR)最小容量阈值。干预后,采用HE染色观察各组大鼠结肠组织形态,扫描电镜观察结肠黏膜超微结构,透射电镜观察结肠黏膜肥大细胞超微结构,甲苯胺蓝染色观察结肠黏膜肥大细胞脱颗粒情况,ELISA法检测大鼠血清和结肠组胺、白细胞介素(IL)-1β、IL-6、IL-1α、类胰蛋白酶、蛋白酶激活受体2(PAR-2)含量,Western blot法和实时荧光定量PCR法检测大鼠结肠IL-1β、IL-6、IL-1α、类胰蛋白酶、PAR-2蛋白和mRNA表达,免疫荧光法检测大鼠结肠组胺、IL-1β、IL-6、IL-1α、类胰蛋白酶、PAR-2阳性表达。结果:与正常组比较,模型组大鼠结肠组织被大量炎性细胞浸润,结肠黏膜严重破坏,绒毛排列散乱,肥大细胞内颗粒电子密度降低、颗粒数量明显减少;稀便率、肥大细胞脱颗粒率升高(P<0.01),AWR最小容量阈值降低(P<0.01);血清和结肠组胺、IL-1β、IL-6、IL-1α、类胰蛋白酶、PAR-2含量升高(P<0.01);结肠组胺阳性表达升高(P<0.01),IL-1β、IL-6、IL-1α、类胰蛋白酶、PAR-2蛋白、m RNA及阳性表达均升高(P<0.01)。与模型组比较,酮替芬组、艾灸组、灸药组大鼠结肠组织炎性细胞浸润明显减少,结肠黏膜较完整,绒毛排列较整齐,肥大细胞内颗粒�Objective To observe the effects of moxibustion on colonic mast cell degranulation and inflammatory factor expression in rats with diarrhea-predominant irritable bowel syndrome(IBS-D),and explore the potential mechanism of moxibustion in treating IBS-D.Methods Forty-five rat pups born from 5 healthy SPF-grade pregnant SD rats,with 8 rats were randomly selected as the normal group.The remaining 37 rats were intervened with maternal separation,acetic acid enema,and chronic restraint stress to establish the IBS-D model.The successfully modeled 32 rats were then randomly assigned to a model group,a ketotifen group,a moxibustion group,and a moxibustion-medication group,with 8 rats in each group.The rats in the ketotifen group were intervened with intragastric administration of ketotifen solution(10 mL/kg);the rats in the moxibustion group were intervened with suspended moxibustion on bilateral"Tianshu"(ST 25)and"Shangjuxu"(ST 37);the rats in the moxibustion-medication group were intervened with suspended moxibustion combined with intragastric administration of ketotifen solution.All interventions were administered once daily for 7 consecutive days.The diarrhea rate and minimum volume threshold of abdominal withdrawal reflex(AWR)were calculated before and after modeling,as well as after intervention.After intervention,colonic tissue morphology was observed using HE staining;colonic mucosal ultrastructure was examined by scanning electron microscopy;colonic mast cell ultrastructure was observed using transmission electron microscopy;mast cell degranulation was assessed by toluidine blue staining;serum and colonic levels of histamine,interleukin(IL)-1β,IL-6,IL-1α,trypsin-like enzyme,and protease-activated receptor 2(PAR-2)were measured by ELISA;the Western blot and real-time quantitative PCR were employed to evaluate the protein and mRNA expression of colonic IL-1β,IL-6,IL-1α,trypsin-like enzyme,and PAR-2;the immunofluorescence was used to detect the positive expression of histamine,IL-1β,IL-6,IL-1α,trypsin-like en

关 键 词：腹泻型肠易激综合征 艾灸 肥大细胞 脱颗粒 低度炎性反应 组胺 类胰蛋白酶 

分 类 号：R245.81[医药卫生—针灸推拿学]