Li Yanyun,Liu Xuelai,Cui Zhao,et al.Histological observation of TLR4/MyD88 signaling pathway in septum arisen from type Ⅰ intestinal atresia[J].Journal of Clinical Pediatric Surgery,2023,22(02):180-184.[doi:10.3760/cma.j.cn101785-202202018-015]
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
- Title:
- Histological observation of TLR4/MyD88 signaling pathway in septum arisen from type Ⅰ intestinal atresia
- Keywords:
- Intestinal Atresia; Etiology; Intestinal Atresia; Pathology; Septum; Immunology; Toll-like Receptor Signaling Pathway
- 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0.6 316±0.0 268,0.5 066±0.0 378,0.6 091±0.0 270,0.3 841±0.0 610,正常肠壁分别为0.4 333±0.0 409,0.3 758±0.0 365,0.4 108±0.0 263,0.2 108±0.0 573,P值分别为0.0 042,0.0 360,0.0 057,0.0 137。 结论 TLR4/MyD88信号通路参与了Ⅰ型肠闭锁隔膜组织的形成或发展。
- Abstract:
- Objective To observe the expressions of TRIF,TRAF3,IFN-β and 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-β and 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-β and 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 0.6 316±0.0 268,0.5 066±0.0 378,0.6 091±0.0 270,0.3 841±0.0 610;normal intestinal wall 0.4 333±0.0 409,0.3 758±0.0 365,0.4 108±0.0 263,0.2 108±0.0 573,p value point ratio 0.0042,0.0 360,0.0 057,0.0 137).Conclusion TLR4/TRIF signaling pathway is involved in formation and development of septum tissue in type I intestinal atresia during fetal period.
参考文献/References:
[1] Sahasrabudhe NM,Dokter-Fokkens J,de Vos P.Particulate β-glucans synergistically activate TLR4 and Dectin-1 in human dendritic cells[J].Mol Nutr Food Res,2016,60(11):2514-2522.DOI:10.1002/mnfr.201600356.
[2] Yao SY,Xu MK,Li YS,et al.Staphylococcal enterotoxin C2 stimulated the maturation of bone marrow derived dendritic cells via TLR-NFκB signaling pathway[J].Exp Cell Res,2018,370(2):237-244.DOI:10.1016/j.yexcr.2018.06.024.
[3] 刘雪来,曹学会,宋岩彪,等.先天性小肠Ⅰ型闭锁胚胎期再贯通残留液泡的组织学形态观察[J].中华小儿外科杂志,2019,40(3):241-244.DOI:10.3760/cma.j.issn.0253-3006.2019.03.010. Liu XL,Cao XH,Song YB,et al.Histological observations of septum from type Ⅰ intestinal atresia and residual vacuoles during recanalization in fetal period[J].Chin J Pediatr Surg,2019,40(3):241-244.DOI:10.3760/cma.j.issn.0253-3006.2019.03.010.
[4] 刘雪来,李龙,谢向辉,等.TLR4、MyD88、TRAF6和IRAK4分子在Ⅰ型小肠闭锁隔膜组织中表达的研究[J].中华小儿外科杂志,2021,42(11):1020-1025.DOI:10.3760/cma.j.cn421158-20200307-00148. Liu XL,Li L,Xie XH,et al.Expressions of TLR4,MyD88,TRAF6 and IRAK4 in septum derived from type I intestinal atresia[J].Chin J Pediatr Surg,2021,42(11):1020-1025.DOI:10.3760/cma.j.cn421158-20200307-00148.
[5] Adams SD,Stanton MP.Malrotation and intestinal atresias[J].Early Hum Dev,2014,90(12):921-925.DOI:10.1016/j.earlhumdev.2014.09.017.
[6] Langer JC.Intestinal rotation abnormalities and midgut volvulus[J].Surg Clin North Am,2016,97(1):147-159.DOI:10.1016/j.suc.2016.08.011.
[7] Morris G,Kennedy A Jr,Cochran W.Small bowel congenital anomalies:a review and update[J].Curr Gastroenterol Rep,2016,18(4):16.DOI:10.1007/s11894-016-0490-4.
[8] Batra A,Keys SC,Johnson MJ,et al.Epidemiology,management and outcome of ultrashort bowel syndrome in infancy[J].Arch Dis Child Fetal Neonatal Ed,2017,102(6):F551-F556.DOI:10.1136/archdischild-2016-311765.
[9] Rigante D.New mosaic tiles in childhood hereditary autoinflammatory disorders[J].Immunol Lett,2018,193:67-76.DOI:10.1016/j.imlet.2017.11.013.
[10] Mohan S,Gupta D.Crosstalk of toll-like receptors signaling and Nrf2 pathway for regulation of inflammation[J].Biomed Pharmacother,2018,108:1866-1878.DOI:10.1016/j.biopha.2018.10.019.
[11] Kim S,Joe Y,Surh YJ,et al.Differential regulation of toll-like receptor-mediated cytokine production by unfolded protein response[J].Oxid Med Cell Longev,2018,2018:9827312.DOI:10.1155/2018/9827312.
[12] Pang Z,Junkins RD,Raudonis R,et al.Regulator of calcineurin 1 differentially regulates TLR-dependent MyD88 and TRIF signaling pathways[J].PLoS One,2018,13(5):e0197491.DOI:10.1371/journal.pone.0197491.
[13] 肖尚杰,杨文熠,朱小春,等.腹腔镜治疗54例新生儿先天性十二指肠梗阻的疗效分析[J].临床小儿外科杂志,2019,18(2):141-146.DOI:10.3969/j.issn.1671-6353.2019.02.014. Xiao SJ,Yang WY,Zhu XC,et al.Analysis of laparoscopic diagnosis and treatment for congenital duodenal obstruction in neonates:a report of 54 cases[J].J Clin Ped Sur,2019,18(2):141-146.DOI:10.3969/j.issn.1671-6353.2019.02.014.
相似文献/References:
[1]张生 金先庆 李晓庆 周德凯 向丽 刘伟. 先天性肠闭锁、肠狭窄120例临床分析[J].临床小儿外科杂志,2011,10(04):273.
[J].Journal of Clinical Pediatric Surgery,2011,10(02):273.
[2]李光源 李婉冰 高梅玲. 肠闭锁产前诊断与产后诊疗21例体会[J].临床小儿外科杂志,2012,11(03):234.
[J].Journal of Clinical Pediatric Surgery,2012,11(02):234.
[3]夏仁鹏 李碧香 周崇高. 先天性结肠闭锁18例诊治分析[J].临床小儿外科杂志,2014,13(05):412.
[4]吴典明 崔旭 林宇. 先天性肠闭锁113例预后分析[J].临床小儿外科杂志,2014,13(06):492.
[5]王娟,丁敏,印其友. PICC在新生儿先天性肠闭锁围手术期的应用[J].临床小儿外科杂志,2015,14(06):555.
[6]张镟,邓涛,张九鸿,等.儿童干燥闭塞性阴茎头炎 8 例及文献复习[J].临床小儿外科杂志,2018,17(07):514.
Zhang Xuan,Deng Tao,Zhang Jiuhong,et al.Balanitis xerotica obliterans in boys:a report of 8 cases and literature review.[J].Journal of Clinical Pediatric Surgery,2018,17(02):514.
[7]王建峰,朱慧,陈杰.肠道间质细胞参与调控结肠动力的研究进展[J].临床小儿外科杂志,2022,21(12):1191.[doi:10.3760/cma.j.cn101785-202010023-017]
Wang Jianfeng,Zhu Hui,Chen Jie.Recent advances in gastrointestinal interstitial cells regulating colonic motility[J].Journal of Clinical Pediatric Surgery,2022,21(02):1191.[doi:10.3760/cma.j.cn101785-202010023-017]
[8]李博雅,李娇,杨博,等.存在治疗抵抗因素的单症状性夜遗尿儿童临床表现及治疗效果分析[J].临床小儿外科杂志,2023,22(02):105.[doi:10.3760/cma.j.cn101785-202212013-002]
Li Boya,Li Jiao,Yang Bo,et al.Clinical symptoms and treatment outcomes of children with monosymptomatic nocturnal enuresis having treatment-resistant factors[J].Journal of Clinical Pediatric Surgery,2023,22(02):105.[doi:10.3760/cma.j.cn101785-202212013-002]
[9]傅润熹,王阳,蔡威.DLL3基因遗传多态性与先天性巨结肠易感性的相关性分析[J].临床小儿外科杂志,2023,22(04):351.[doi:10.3760/cma.j.cn101785-202203030-010]
Fu Runxi,Wang Yang,Cai Wei.Association study of delta-ligand 3 (DLL3) gene with Hirschsprung’s disease susceptibility[J].Journal of Clinical Pediatric Surgery,2023,22(02):351.[doi:10.3760/cma.j.cn101785-202203030-010]
[10]刘万林,孙亮,赵振群,等.骨化核与发育性髋关节发育不良闭合复位后股骨头缺血坏死相关性的研究进展[J].临床小儿外科杂志,2023,22(06):584.[doi:10.3760/cma.j.cn101785-202010041-017]
Liu Wanlin,Sun Liang,Zhao Zhenqun,et al.Research advances on the relationship of ossification nucleus with avascular necrosis of developmental hip dysplasia after closed reduction[J].Journal of Clinical Pediatric Surgery,2023,22(02):584.[doi:10.3760/cma.j.cn101785-202010041-017]
备注/Memo
收稿日期:2021-8-31。
基金项目:北京市自然科学基金(7222015)
通讯作者:刘雪来,Email:liuxuelai_steven@163.com