中华小儿外科杂志
中華小兒外科雜誌
중화소인외과잡지
CHINESE JOURNAL OF PEDIATRIC SURGERY
2014年
9期
698-702
,共5页
高明娟%刘远梅%毛羽晨%金祝%郑泽兵%曲颜%孔萌
高明娟%劉遠梅%毛羽晨%金祝%鄭澤兵%麯顏%孔萌
고명연%류원매%모우신%금축%정택병%곡안%공맹
消化系统畸形%直肠%肛门%神经生长因子%肠神经系统
消化繫統畸形%直腸%肛門%神經生長因子%腸神經繫統
소화계통기형%직장%항문%신경생장인자%장신경계통
Digestive system abnormalities%Rectum%Anus%Nerve growth factors%Enteric nervous system
目的 观察神经生长因子(nerve growth factor,NGF)及其受体(TrkA、P75NTR)在先天性肛门直肠畸形(anorectal malformation,ARM)患儿直肠末端的表达,探讨对其肠神经系统发育的影响.方法 收集2011年1月至2013年1月遵义医学院附属医院诊治的ARM患儿直肠末端标本30例.其中,男24例,女6例;高位ARM 15例,中位ARM 6例,低位ARM 9例;手术年龄2 d～13个月,平均1.4个月.一期腹会阴肛门成形8例;分期肛门成形13例,包括:腹腔镜辅助肛门成形7例,腹会阴肛门成形2例,后矢状入路肛门成形术4例,会阴肛门成形9例.肛门成形术中取直肠末端标本,运用苏木素/伊红(HE)染色计数直肠末端黏膜下及肌间神经丛数量,免疫组织化学(IHC)及实时定量聚合酶链反应(qRT-PCR)检测NGF及其受体(TrkA、P75NTR)在低位ARM和中高位ARM患儿直肠末端的表达情况.结果 HE染色:低位ARM患儿直肠末端黏膜下及肌间神经丛分布密集,神经丛面积大,节细胞数量多;中高位ARM患儿神经丛分布明显稀疏,神经丛数量较低位ARM患儿明显减少(1.30±0.82vs5.60±1.36)个,两相比较,差异有统计学意义(P＜0.01).IHC:NGF、TrkA、P75NTR主要在直肠末端黏膜下及肌间神经丛表达,NGF主要定位于胞核,TrkA主要定位于胞核及胞质,P75NTR主要定位于胞核、胞膜.NGF、TrkA、P75NTR在低位ARM患儿直肠黏膜下、肌间神经丛大量表达,其积分光密度(IOD值)分别为43.48±7.11、18.83±3.65和23.39±3.64;NGF、TrkA、P75NTR在中高位ARM患儿直肠末端表达明显减少,其积分光密度(IOD值)分别为3.70±1.21、8.49±2.68和7.17±2.28,两相比较,差异均有统计学意义(P＜0.01).qRT PCR:NGF、TrkA、P75NTR mRNA在低位ARM患儿直肠末端的相对表达量分别为(172.22±104.38、148.78±72.45和122.83±35.45);明显高于中高位ARM患儿(80.38±43.10、73.95±47.43和75.15±20.23),两相比较,差异均有统计学意义(P＜0.01).结论 NGF及其受体TrkA、P75NTR可能参与并影响ARM患儿直肠末端肠神经系统的发育.
目的 觀察神經生長因子(nerve growth factor,NGF)及其受體(TrkA、P75NTR)在先天性肛門直腸畸形(anorectal malformation,ARM)患兒直腸末耑的錶達,探討對其腸神經繫統髮育的影響.方法 收集2011年1月至2013年1月遵義醫學院附屬醫院診治的ARM患兒直腸末耑標本30例.其中,男24例,女6例;高位ARM 15例,中位ARM 6例,低位ARM 9例;手術年齡2 d～13箇月,平均1.4箇月.一期腹會陰肛門成形8例;分期肛門成形13例,包括:腹腔鏡輔助肛門成形7例,腹會陰肛門成形2例,後矢狀入路肛門成形術4例,會陰肛門成形9例.肛門成形術中取直腸末耑標本,運用囌木素/伊紅(HE)染色計數直腸末耑黏膜下及肌間神經叢數量,免疫組織化學(IHC)及實時定量聚閤酶鏈反應(qRT-PCR)檢測NGF及其受體(TrkA、P75NTR)在低位ARM和中高位ARM患兒直腸末耑的錶達情況.結果 HE染色:低位ARM患兒直腸末耑黏膜下及肌間神經叢分佈密集,神經叢麵積大,節細胞數量多;中高位ARM患兒神經叢分佈明顯稀疏,神經叢數量較低位ARM患兒明顯減少(1.30±0.82vs5.60±1.36)箇,兩相比較,差異有統計學意義(P＜0.01).IHC:NGF、TrkA、P75NTR主要在直腸末耑黏膜下及肌間神經叢錶達,NGF主要定位于胞覈,TrkA主要定位于胞覈及胞質,P75NTR主要定位于胞覈、胞膜.NGF、TrkA、P75NTR在低位ARM患兒直腸黏膜下、肌間神經叢大量錶達,其積分光密度(IOD值)分彆為43.48±7.11、18.83±3.65和23.39±3.64;NGF、TrkA、P75NTR在中高位ARM患兒直腸末耑錶達明顯減少,其積分光密度(IOD值)分彆為3.70±1.21、8.49±2.68和7.17±2.28,兩相比較,差異均有統計學意義(P＜0.01).qRT PCR:NGF、TrkA、P75NTR mRNA在低位ARM患兒直腸末耑的相對錶達量分彆為(172.22±104.38、148.78±72.45和122.83±35.45);明顯高于中高位ARM患兒(80.38±43.10、73.95±47.43和75.15±20.23),兩相比較,差異均有統計學意義(P＜0.01).結論 NGF及其受體TrkA、P75NTR可能參與併影響ARM患兒直腸末耑腸神經繫統的髮育.
목적 관찰신경생장인자(nerve growth factor,NGF)급기수체(TrkA、P75NTR)재선천성항문직장기형(anorectal malformation,ARM)환인직장말단적표체,탐토대기장신경계통발육적영향.방법 수집2011년1월지2013년1월준의의학원부속의원진치적ARM환인직장말단표본30례.기중,남24례,녀6례;고위ARM 15례,중위ARM 6례,저위ARM 9례;수술년령2 d～13개월,평균1.4개월.일기복회음항문성형8례;분기항문성형13례,포괄:복강경보조항문성형7례,복회음항문성형2례,후시상입로항문성형술4례,회음항문성형9례.항문성형술중취직장말단표본,운용소목소/이홍(HE)염색계수직장말단점막하급기간신경총수량,면역조직화학(IHC)급실시정량취합매련반응(qRT-PCR)검측NGF급기수체(TrkA、P75NTR)재저위ARM화중고위ARM환인직장말단적표체정황.결과 HE염색:저위ARM환인직장말단점막하급기간신경총분포밀집,신경총면적대,절세포수량다;중고위ARM환인신경총분포명현희소,신경총수량교저위ARM환인명현감소(1.30±0.82vs5.60±1.36)개,량상비교,차이유통계학의의(P＜0.01).IHC:NGF、TrkA、P75NTR주요재직장말단점막하급기간신경총표체,NGF주요정위우포핵,TrkA주요정위우포핵급포질,P75NTR주요정위우포핵、포막.NGF、TrkA、P75NTR재저위ARM환인직장점막하、기간신경총대량표체,기적분광밀도(IOD치)분별위43.48±7.11、18.83±3.65화23.39±3.64;NGF、TrkA、P75NTR재중고위ARM환인직장말단표체명현감소,기적분광밀도(IOD치)분별위3.70±1.21、8.49±2.68화7.17±2.28,량상비교,차이균유통계학의의(P＜0.01).qRT PCR:NGF、TrkA、P75NTR mRNA재저위ARM환인직장말단적상대표체량분별위(172.22±104.38、148.78±72.45화122.83±35.45);명현고우중고위ARM환인(80.38±43.10、73.95±47.43화75.15±20.23),량상비교,차이균유통계학의의(P＜0.01).결론 NGF급기수체TrkA、P75NTR가능삼여병영향ARM환인직장말단장신경계통적발육.
Objective To explore the roles of nerve growth factor (NGF) and nerve growth factor receptor (NGFR) (TrkA,P75NTR) in enteric nervous system of distal rectum of children with anorectal malformations (ARM).Methods Specimens were obtained from 30 (24 males,6 females) children's distal rectum with ARM at our hospital during January 2011 to January 2013; According to the position of malformations,there were high (n =15),intermediate (n =6) and low (n =9).Their average age was 1.4 months (range,2 days to 13 months).One-stage abdominal-perineal anoplasty were performed in 8 cases.Among 13 cases of anoplasty by stages,the procedures included laparoscopic-assisted anorectal pull-through (LAARP) (n =7),abdominal-perineal anoplasty (n =2),posterior sagittal anorectoplasty (n=4) and one-stage anpoerineal plasty (n=9).Distal rectum was collected during operation.The hematoxylin-eosin (HE) staining was used to count the number of submucosal plexus and myenteric plexus of distal rectum.And immunohistochemistry (IHC) and quantitative real-time polymerase chain reaction (qRT-PCR) were employed to detect the expressions of NGF and NGFR in distal rectum of patients with ARM.And the expression level of low ARM was compared with that of intermediate or high ARM.Results HE staining:for patients with low ARM,distal rectum submucous and myenteric plexuses were dense with a large plexus area and a large number of intestinal ganglion cell while the distribution of nerve plexus of patients with intermediate or high ARM were obviously few and scattered.Comparing with low ARM,the number of nerve plexus obviously decreased (1.30 ± 0.82 vs.5.60 ± 1.36,P＜0.01) ; IHC:NGF,TrkA and p75NTR were predominantly distributed in distal rectum submucous and myenteric plexuses,NGF & TrkA in nucleus and cytoplasm and P75NTR in nucleus and cell membrane; NGF,TrkA and P75NTR had abundant expression in patients with low ARM in distal rectum submucous and myenteric plexuses,the expression quantity of patients with intermediate or high ARM obviously decreased.The integral optical density (IOD) was (43.48 ±7.11 vs 3.70 ± 1.21; 18.83 ± 3.65 vs 8.49 ±2.68; 23.39 ± 3.64 vs 7.17 ± 2.28,P＜0.01) ; qRT-PCR:The relative expression of NGF,TrkA and P75NTR mRNA at patients with low ARM was obviously higher than that of intermediate or high ARM's (172.22 ± 104.38 vs 80.38 ± 43.10; 148.78 ± 72.45 vs 73.95 ± 47.43; 122.83 ± 35.45 vs 75.15 ± 20.23,P＜ 0.01).Conclusions NGF,TrkA and p75NTR may play an important role in the development of enteric nervous system in distal rectum of children with ARM.
