TLR4/TRIF信号参与Ⅰ型小肠闭锁隔膜组织黏膜免疫应答的组织学研究  

Histological observation of TLR4/MyD88 signaling pathway in septum arisen from type Ⅰ intestinal atresia

作　　者：李艳云 刘雪来[2] 崔钊[3] 高鹏 宋岩彪 Li Yanyun;Liu Xuelai;Cui Zhao;Gao Peng;Song Yanbiao(Department of Neonatology,Bayi Children's Hospital,Seventh Medical Center of PLA General Hospital,Beijing 100700,China;Department of Surgery,Capital Institute of Pediatrics Affiliated Children's Hospital,Beijing 100020,China;Department of Surgery,Municipal Children's Hospital,Chang Chun 130061,China;Department of Surgery,Harbin Children's Hospital,Harbin 150010,China;Department of Central Labarotory,Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]解放军总医院儿科医学部,解放军总医院第七医学中心儿科研究所,出生缺陷防控关键技术国家工程实验室,儿童器官功能衰竭北京市重点实验室,北京100700 [2]首都儿科研究所附属儿童医院外科,北京100020 [3]长春市儿童医院外科,长春130061 [4]哈尔滨市儿童医院外科,哈尔滨150010 [5]河北医科大学第二医院中心实验室,石家庄050000

出　　处：《临床小儿外科杂志》2023年第2期180-184,共5页Journal of Clinical Pediatric Surgery

基　　金：北京市自然科学基金(7222015)

摘　　要：目的观察Ⅰ型小肠闭锁肠腔隔膜组织黏膜β干扰素TIR结构域衔接蛋白(TIR-domain-containing adapter-inducing interferon-β,TRIF)、肿瘤坏死因子受体相关因子蛋白3(tumor receptor associated factor,TRAF3)、干扰素-β(interferon-beta,IFN-β)和干扰素-λ1(interferon-λ1,IFN-λ1)分子的表达特征,探索胚胎期Toll样受体4(toll like receptor 4,TLR4)-TRIF非依赖性信号通路与肠管再通过程的相关性。方法采用免疫组化技术观察和比较新生儿正常肠壁和Ⅰ型肠闭锁隔膜组织黏膜层内TRIF通路内重要分子的表达特征。收集16例患儿组织标本(解放军总医院儿科医学部1例,首都儿科研究所1例,河北医科大学第二医院6例,哈尔滨市儿童医院5例,长春市儿童医院3例),均为术中证实的Ⅰ型肠闭锁隔膜组织,为隔膜组;以同一患儿术中行肠切除、肠吻合过程中钳取收集的正常肠壁组织为对照组。患儿手术年龄为出生后1~3 d,其中空肠闭锁13例(男5例,女8例),回肠闭锁3例(男1例,女2例)。将组织标本转入快速组织处理系统进行组织标本连续切片。采用常规HE染色进行定位,隔膜组和对照组各16例,每个病例染色后随机取4个视野(非肌肉区域),应用图像分析软件Image pro plus 6.0测量积分光密度(integral-optical density,IOD)和面积(Area),并计算出平均吸光度(A)(OD)。OD=IOD/Area。采用t检验和双盲法分析免疫组化平均光密度。结果新生儿正常肠壁组织黏膜层的TRIF、TRAF3、IFN-β和IFN-λ1的分子表达不明显或低表达,在隔膜组织黏膜层表达显著。半定量分析结果显示,TLR4、MyD88、TRAF6和IRAK4分子在隔膜组织黏膜层表达比新生儿正常肠壁黏膜层多,二者差异有统计学意义(隔膜组织分别为0.6316±0.0268,0.5066±0.0378,0.6091±0.0270,0.3841±0.0610,正常肠壁分别为0.4333±0.0409,0.3758±0.0365,0.4108±0.0263,0.2108±0.0573,P值分别为0.0042,0.0360,0.0057,0.0137)。结论TLR4/MyD88信号通路参与了Ⅰ�Objective To observe the expressions of TRIF,TRAF3,IFN-βand IFN-λ1 in septum derived from type I intestinal atresia and examine their correlations.Methods Immunohistochemistry was utilized for observing and comparing the expressions of important molecules in TRIF pathway in mucosa of normal intestinal wall and type I intestinal atresia diaphragm.A total of 16 tissue samples were collected from Department of Pediatric Medicine,General Hospital of PLA(n=1),Capital Institute of Pediatrics(n=1),Second Hospital,Hebei Medical University(n=6),Harbin Children's Hospital(n=5)and Changchun Children's Hospital(n=3).All samples were intraoperatively confirmed as type I intestinal atresia diaphragm tissue(diaphragm group).Normal intestinal wall tissues collected during intestinal resection and anastomosis in the same child were selected as control group.Totally 16 septa in operated neonates at post-birth 1~3 days were collected,including 13 septa(male 5,female 8)derived from jejunum and 3 septa(male 1,female 2)from ileum.Normal intestinal wall tissues were collected as control during surgery in the same neonate.All fresh tissues were processed for serial tissue sectioning and hematoxylin-eosin(HE)&immunohistochemical stains of TRIF,TRAF3,IFN-βand IFN-λ1 molecules in each group for histological observations.Four visual fields under immunohistochemical stain were targeted randomly for semi-quantitative calculation of average optical density(OD)by Image pro plus6.0 software.Statistical methods of t test and double blinding were employed for analyzing the average optical density of immunohistochemistry.Results Immunohistochemical stain indicated that the expressions of TRIF,TRAF3,IFN-βand IFN-λ1 in septum tissue were significant compared with those in normal neonatal intestinal wall.Semi-quantitative calculation indicated marked elevations in septum tissue in contrast to those in normal neonatal intestinal wall(fraction of diaphragm tissue was 0.6316±0.0268,0.5066±0.0378,0.6091±0.0270,0.3841±0.0610;normal intestinal wal

关 键 词：肠闭锁 病因学 肠闭锁 病理学 隔膜组织 免疫学 Toll样信号通路 

分 类 号：R726.5[医药卫生—儿科]