艾灸“天枢”穴对溃疡性结肠炎模型大鼠结肠组织铁死亡及氧化损伤的影响  

Effects of Moxibustion at“Tianshu”(ST25)on Ferroptosis and Oxidative Damage in Colon Tissue of Ulcerative Colitis Model Rats

作　　者：刘力 卢云琼 曹姚佳妮 李红娜 金悠悠[2] 孔谐和 樊晓鹏[1] 洪珏[3] 马晓芃[3] 张丹[3] LIU Li;LU Yunqiong;CAO Yaojiani;LI Hongna;JIN Youyou;KONG Xiehe;FAN Xiaopeng;HONG Jue;MA Xiaopeng;ZHANG Dan(Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medi-cine,Shanghai,200437;Kunshan Hospital of Chinese Medicine,Jiangsu;Shanghai Research Institute of Acupuncture and Meridian)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院,上海市200437 [2]江苏省昆山市中医医院 [3]上海市针灸经络研究所

出　　处：《中医杂志》2023年第15期1576-1584,共9页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82104985);上海市自然科学基金(19ZR1451600,23ZR1460100);上海市领军人才项目(060)。

摘　　要：目的 探讨艾灸天枢穴治疗溃疡性结肠炎(UC)的可能作用机制。方法 将42只SD大鼠随机分为正常组9只和造模组33只。造模组采用自由饮用3.5%葡聚糖硫酸钠盐(DSS)溶液7天制备UC大鼠模型。32只造模成功,将其随机分为模型组、艾灸组、铁死亡抑制剂组、溶媒对照组,每组8只。正常组和模型组只做与干预组相同的抓取固定;艾灸组取“天枢穴”(双侧)进行温和灸,每次每穴10 min;铁死亡抑制剂组采用浓度为6 mg/ml的铁死亡抑制剂利普司他丁-1 (Liproxstatin-1)以10 mg/kg腹腔注射;溶媒对照组给予5%二甲基亚砜(DMSO) 1.67 ml/kg腹腔注射。各组均每日1次,连续8天。干预后进行结肠大体形态损伤指数(CMDI)评分,HE染色观察大鼠结肠组织形态结构变化,酶联免疫吸附测定法检测血清二胺氧化酶(DAO)表达,微板法检测血清D-乳酸(D-LA)含量,TUNEL染色观察结肠细胞死亡情况,免疫荧光法检测结肠组织活性氧(ROS)表达,比色法检测结肠铁离子的含量,免疫组织化学法检测结肠谷胱甘肽过氧化物酶4 (GPX4)表达。结果 与正常组比较,模型组与溶媒对照组大鼠结肠结构破坏严重,CMDI评分升高(P<0.01),HE染色可见浅表性溃疡,黏膜层破坏明显,黏膜下层大量炎性细胞浸润和水肿,结肠细胞死亡数增多,血清DAO、D-LA及结肠组织铁离子、ROS含量均增多,结肠组织GPX4蛋白表达减少(P<0.05或P<0.01)。与模型组比较,艾灸组、铁死亡抑制剂组大鼠结肠黏膜、腺体有不同程度修复,肠道炎症减轻,细胞死亡数显著减少,CMDI评分明显降低,血清DAO、D-LA及结肠组织ROS、铁离子含量均明显减少,结肠组织GPX4蛋白表达增多(P<0.05或P<0.01)。结论 艾灸“天枢穴”能降低结肠组织铁离子和ROS含量,上调GPX4蛋白表达,抑制铁死亡,可能是其促进UC结肠黏膜愈合、治疗UC的作用机制之一。Objective To explore the possible mechanism of moxibustion at tianshu(ST 25)in treating ulce-rative colitis(UC).Methods Forty-two SD rats were randomly divided into normal(n=9)and modeling group(n=33).UC model was prepared by freely drinking of 3.5%dextran sulfate sodium salt(DSS)solution for 7 days in the modeling group.Thirty-two successfully modeled rats were randomly divided into model group,moxibustion group,fer⁃roptosis inhibitor group,and vehicle control group,with eight rats in each group.The normal and model groups only did the same grasping fixation as the intervention groups;the moxibustion group was treated with mild moxibustion at the bilateral tianshu(ST 25)for 10 min each time;the ferroptosis inhibitor group was intervened with 6 mg/ml of fer⁃roptosis inhibitor liproxstatin-1 by intraperitoneal injection(10 mg/kg);and the vehicle group was given 1.67 ml/kg of 5%dimethyl sulfoxide(DMSO)intraperitoneally.Each group was treated once a day for 8 days.After the intervention,the Colon Gross Morphological Damage Index(CMDI)score was evaluated.HE staining was used to observe the morphological and structural changes of rat colon tissue.The serum diamine oxidase(DAO)expression was detected by enzyme-linked immunosorbent assay(ELISA),while the content of D-lactic acid(D-LA)in the serum was deter⁃mined by the microplate method.The colonic cell death was observed by TUNEL staining,and the expression of reac⁃tive oxygen species(ROS)in the colon was detected by immunofluorescence.The content of colonic Fe ions was tested by the colorimetric method,and the expression of lutathione peroxidase 4(GPX4)in the colon was detected by immu⁃nohistochemistry.Results Compared to those in the normal group,the colonic tissue damage of rats in the model group and vehicle control group was serious,with an increased CMDI score(P<0.01);visible superficial ulcers,ob⁃vious destruction of the mucosal layer,and a large number of inflammatory cells infiltrating the submucosal layer alongside edema were found;the colonic cel

关 键 词：溃疡性结肠炎 艾灸 天枢穴 铁死亡 氧化损伤 

分 类 号：R245[医药卫生—针灸推拿学]