TLR4、MyD88、TRAF6和IRAK4分子在Ⅰ型小肠闭锁隔膜组织中表达的研究  

Expressions of TLR4, MyD88, TRAF6 and IRAK4 in septum derived from type I intestinal atresia

作　　者：刘雪来[1] 李龙[1] 谢向辉 宋岩彪 杜娟[3] 李索林[4] 李英超[4] 高鹏 崔钊[6] Liu Xuelai;Li Long;Xie Xianghui;Song Yanbiao;Du Juan;Li Suolin;Li Yingchao;Gao Peng;Cui Zhao(Department of Surgery,Capital Institute of Pediatrics affiliated Children Hospital,Beijing 100020,China;Department of Central Labarotory,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Endocrinology,Jilin Provincial People's Hospital,Changchun 130021,China;Department of Pediatric Surgery,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Surgery,Harbin Children Hospital,Harbin 150010,China;Department of Surgery,Changchun Children'Hospital,Changchun 130061,China)

机构地区：[1]首都儿科研究所附属儿童医院外科,北京100020 [2]河北医科大学第二医院中心实验室,石家庄050000 [3]吉林省人民医院内分泌科,长春130021 [4]河北医科大学第二医院小儿外科,石家庄050000 [5]哈尔滨市儿童医院外科,150010 [6]长春市儿童医院外科,130061

出　　处：《中华小儿外科杂志》2021年第11期1020-1025,共6页Chinese Journal of Pediatric Surgery

基　　金：北京市属医学科研院所公益发展改革试点项目 (京医研2019-11)。

摘　　要：目的观察Ⅰ型小肠闭锁肠腔隔膜组织黏膜Toll样受体4(toll-like receptor-4,TLR4)和髓样分化蛋白88(myeloid differential protein-88,MyD88)信号通路分子表达,并探索胚胎期TLR4/MyD88依赖性信号通路与肠管再通过程的相关性。方法收集16例术中证实为Ⅰ型小肠闭锁患儿的肠腔隔膜组织。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);手术年龄为出生后1~3 d。以同一患儿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。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,采用免疫组织化学染色和观察TLR4和MyD88,以及该信号通路下游的肿瘤坏死因子受体相关分子6(tumor necrosis factor receptor-associated factor 6,TRAF6)和白细胞介素-1受体相关激酶4(interleukin-1 receptor associated kinase-4,IRAK4)在正常新生儿肠壁组织黏膜层内、隔膜组织黏膜层内的表达。各组分别随机取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(IOD)值和面积(Area)并计算出平均光密度(OD)值。OD=IOD/Area。采用双盲法分析比较免疫组织化学平均光密度值并进行统计学分析。结果免疫组织化学结果显示,正常肠壁组织黏膜层TLR4、MyD88、TRAF6和IRAK4分子表达不明显,但上述指标在隔膜组织黏膜层表达显著。半定量结果显示,TLR4、MyD88、TRAF6和IRAK4分子在新生儿正常肠壁黏膜层的相对表达量分别为0.010、0.0122、0.1008和0.0102,而其在隔膜组织黏膜层的相对表达量分别为0.048、0.1072、0.0887和0.0468,两相比较,差异均有统计学意义(P=0.001、0.0046、0.0016和0.001)。结论Ⅰ型肠闭锁隔膜黏膜层TLR4、MyD88、TRAF6和IRAK4分子表达相对正常肠壁组织增多,提示TLR4/MyD88信号通路可能与Ⅰ型肠闭锁隔膜组织的形成或发展具有相关性。Objective To explore the expressions of molecules correlated with toll-like receptor-4(TLR4),myeloid differential protein-88(MyD88)signaling pathway in septum derived from type I intestinal atresia and to examine its correlation.Methods A total of 16 septa in surgical neonates at postnatal 1~3 days were collected,including 13 septum(male 5,female 8)derived from jejunum and 3 septum(male 1,female 2)from ileum.Normal intestinal wall tissues were collected intraoperatively as control in the same neonate.All fresh tissues were processed for serial tissue sectioning,hematoxylin-eosin(HE)stain and immunohistochemical analysis of TLR4/MyD88,down-stream tumor necrosis factor receptor-associated factor 6(TRAF6)and interleukin-1 receptor associated kinase-4(IRAK4).Four random visual fields under immunohistochemical staining were targeted for semi-quantitative calculation of average optical density(OD)by Image pro plus6.0 software,followed by double-blind statistical analysis and comparison.Results Macroscopic observation indicated that the expressions of TLR4,MyD88,TRAF6 and IRAK4 were significant in mucous layers of septum tissue as compared with those in normal neonatal intestinal wall.Semi-quantitative calculation indicated marked elevations in mucous layers of septum tissue in contrast to those in normal neonatal intestinal wall(TLR4 in normal neonatal intestinal wall∶TLR4 in septum tissue=0.048∶0.010,P=0.001;MyD88 in normal neonate intestinal wall∶MyD88 in septum tissue=0.1072∶0.0122,P=0.0046;TRAF6 in normal neonatal intestinal wall∶TRAF6 in septum tissue=0.0887∶0.1008,P=0.0016;IRAK4 in normal neonatal intestinal wall∶IRAK4 in septum tissue=0.0468∶0.0102,P=0.001).Conclusions Up-regulations of TLR4,MyD88,TRAF6 and IRAK4 hint that TLR4/MyD88 signaling pathway in mucous layers is involved in the formation and development of septum tissue during fetal period.

关 键 词：小肠闭锁 隔膜组织 黏膜层 信号通路 

分 类 号：R725.7[医药卫生—儿科]