粪菌移植对不同溃疡性结肠炎小鼠的疗效及其中药性味研究  

Medicinal Flavor of Fecal Microbiota and Efficacies of Its Transplantation in Mice with Different Types of Ulcerative Colitis

作　　者：马晓飞 胡家丽 崔曼曼 葛巍[2] 杨苏琴[2] 丁成华[3] 张磊昌[2] MA Xiaofei;HU Jiali;CUI Manman;GE Wei;YANG Suqin;DING Chenghua;ZHANG Leichang(Graduate School,Jiangxi University of Chinese Medicine,Nanchang 330004,China;Affiliated Hospital of Jiangxi University of Chinese Medicine,Nanchang 330046,China;School of Basic Medical Sciences,Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]江西中医药大学研究生院,江西省南昌市330004 [2]江西中医药大学附属医院,江西省南昌市330046 [3]江西中医药大学基础医学院,江西省南昌市330004

出　　处：《中国全科医学》2022年第15期1875-1882,共8页Chinese General Practice

基　　金：国家自然科学基金资助项目(81760871);江西省教育厅课题(GJJ190682);江西省自然科学基金资助项目(2020BABL206135)。

摘　　要：背景溃疡性结肠炎被世界卫生组织列为现代难治性疾病之一。目前治疗技术均有相应的不足之处,粪菌移植(FMT)治疗溃疡性结肠炎的有效性已被证实,但疗效参差不齐。目的比较FMT治疗小鼠普通型溃疡性结肠炎模型(CUCM)与湿热型溃疡性结肠炎模型(DUCM)的效果及FMT(新金汁)的中药性味。方法于2019年12月9—28日,将35只雄性SPF级C57BL/6小鼠分为7组,分别为正常对照组(Control组)、CUCM组、CUCM+FMT组、CUCM+5-氨基水杨酸(5-ASA)组、DUCM组、DUCM+FMT组、DUCM+5-ASA组,每组各5只小鼠。Control组不做任何干预处理;CUCM组和DUCM组根据造模需求分别制备模型;CUCM+FMT组和DUCM+FMT组于造模成功后,给予制备的粪菌液0.2 ml对小鼠进行灌肠;CUCM+5-ASA组和DUCM+5-ASA组于造模成功后,给予0.0195 g/ml 5-ASA对小鼠进行灌肠。通过HE染色观察各组肠组织的变化,透射电镜观察组织的超微结构变化,流式细胞检测辅助性T细胞1(Th1)、Th2细胞含量,血常规检测血液中白细胞计数(WBC)、红细胞计数(RBC)、血小板计数(PLT)和血红蛋白(HGB),尼莫地平法评估干预前后疗效指数,16S rRNA高通量测序技术检测肠内容物中的菌群分布。结果各组小鼠均造模成功。肠组织HE染色显示:CUCM组与DUCM组肠黏膜表面出现不同程度的缺损或脱落坏死;CUCM+FMT组、CUCM+5-ASA组、DUCM+FMT组、DUCM+5-ASA组肠黏膜基本完整;并且DUCM+FMT组与CUCM+FMT组相比,前者腺体较后者排列整齐并紧密。肠组织透射电镜超微结构显示:CUCM组与DUCM组上皮细胞表面微绒毛稀疏,杯状细胞减少;CUCM+FMT组、CUCM+5-ASA组、DUCM+FMT组、DUCM+5-ASA组微绒毛较为致密,杯状细胞数目较多;并且DUCM+FMT组与CUCM+FMT组相比,前者微绒毛较后者紧密,杯状细胞也较后者多。各组小鼠Th1、Th2细胞含量、WBC、RBC、PLT、HGB比较,差异均有统计学意义(P<0.001)。CUCM+5-ASA组、DUCM+FMT组、DUCM+5-ASA组疗效指数高于CUCM+FMT组(P<0.0Background As an intractable disease recognized by the WHO,current available treatments have limitations for ulcerative colitis(UC).Although the effectiveness of fecal microbiota transplantation(FMT)has been confirmed,the reported efficacies are various.Objective To compare the effect of FMT between a mouse model of common UC and a mouse model of damp-heat UC,and the medicinal flavor of fecal microbiota(new jinzhi).Methods This experiment was implemented from December 9th to 28th,2019.Thirty-five male SPF C57BL/6 mice were equally divided into seven groups:normal control group(no intervention),common UC model(CUCM)group(received interventions to be a CUCM according to the requirements),CUCM+FMT group(received interventions to be a CUCM successfully,then received an enema of 0.2 ml prepared fecal microbiota solution),CUCM+5-aminosalicylic acid(5-ASA)group(received interventions to be a CUCM successfully,then received an enema of 0.0195 g/ml 5-ASA),damp-heat UC model(DUCM)group(received interventions to be a DUCM according to the requirements),DUCM+FMT group(received interventions to be a DUCM successfully,then received an enema of 0.2 ml prepared fecal microbiota solution),and DUCM+5-ASA group(received interventions to be a DUCM successfully,then received an enema of 0.0195 g/ml 5-ASA).The intestinal tissues of each group of mice were taken out when the experiment ended,and were stained with H&E for observing the morphology,and ultrastructure of them was observed using transmission electron microscope,Th1 and Th2 cells in them were detected using flow cytometry,and microbial community in them were detected using high-throughput 16S rRNA gene sequencing.Routine blood test was conducted to measure the levels of white blood cell count(WBC),red blood cell count(RBC),platelet count(PLT)and hemoglobin(HGB).Nimodipine method was used to evaluate the efficacy index before and after intervention.Results The mice in each group were successfully established.The morphology of intestinal tissues stained with H&E:the intestinal

关 键 词：结肠炎 溃疡性 粪菌移植 5-氨基水杨酸 菌群多样性 Th1-Th2平衡 新金汁 

分 类 号：R574.62[医药卫生—消化系统]