Notch受体、配体在胆道发育不良患儿肝纤维化中的表达及意义

Expression and significance of Notch receptors and ligands in liver fibrosis of pediatric biliary dysplasia

目的探讨Notch信号通路蛋白在胆道发育不良(biliary dysplasia,BD)肝纤维化进程中的表达及意义。方法选取2015年1月到2019年1月天津市儿童医院和天津市第一中心医院收治的BD患儿(BD组)19例,术中取肝组织标本;同时收集10例正常肝组织样本(因非肝胆系统疾病死亡患儿)作为对照组。采用免疫组化方法检测各组肝组织中Notch受体和配体的表达。此外,按Ohkuma's肝纤维化分级标准,将BD组患儿分为2个亚组(0~Ⅱ级轻度纤维化组,A组,10例),(Ⅲ~Ⅳ级重度纤维化组,B组,9例);同时将BD组中年龄>180 d的10例患儿,按同样的标准分为轻度纤维化亚组(C组,5例)和重度纤维化亚组(D组,5例),分别比较各亚组间上述蛋白表达的差异。两组比较采用t检验。结果①免疫组化定性结果:Notch1在对照组和BD组中皆为阳性表达;Notch2、3及Jagged1在对照组均为弱阳性表达,而在BD组均为阳性表达。Notch4、Jagged2在各组中表达均为阴性。②免疫组化半定量结果:B组Notch3及Jagged1的表达水平均显著高于A组,差异均有统计学意义(P<0.05);而C组和D组患儿中,Notch2在D组表达水平低于A组,差异具有统计学意义(P<0.05)。结论Notch3及Jagged1的表达会随着肝纤维化程度的加重而升高,提示Notch3和Jagged1可能参与了胆道发育不良肝纤维化进程。在年龄>180 d胆道发育不良患儿中,Notch2的表达会随肝纤维化程度的加重而下降,提示其与胆道发育不良肝纤维化进展缓慢相关。

Objective To explore the expression and significance of Notch signaling pathway in the process of liver fibrosis in pediatric biliary dysplasia(BD).Methods From January 2015 to January 2019,19 BD children were selected.Liver tissue samples were harvested intraoperatively.And 10 normal liver tissue samples collected at the same time(autopsy of deceased patients of non-hepatobiliary diseases)served as control group.The expressions of Notch receptor(Notch1-4)and Notch ligand(Jagged 1/2)in liver tissues of each group were detected by immunohistochemical staining.In addition,children in BD group were divided into 2 subgroups according to the Ohkuma's grading criteria of liver fibrosis(0-2 grade:mild fibrosis group,n=10;3-4 grade:severe fibrosis group,n=9);At the same time,10 children in BD group aged>180 days were divided into the subgroups of mild fibrosis(n=5)and severe fibrosis(n=5)according to the same criteria.The differential expressions of the above proteins between subgroups were examined.Results Immunohistochemical staining results:Notch1 was positively expressed in both control and BD groups;Notch2,3 and Jagged1 were weakly positive in control group,but positive in BD group.Notch4 and Jagged2 were negative in each group.Immunohistochemical semi-quantitative results:The overall expressions levels of receptor of Notch3 and Jagged1 were higher in BD group than those in control group,and the expressions of Notch3 and Jagged1 were higher in severe fibrosis group than those in mild fibrosis group(P<0.05)and the difference was statistically significant(P<0.05).The expression level of Notch2 was lower in severe fibrosis group than that in mild fibrosis group.And the difference was statistically significant(P<0.05).Conclusions The expresssions of Notch3 and Jagged1 were up-regulated with the worsening degree of hepatic fibrosis.It suggested that Notch3 and Jagged1 might participated in the progression of hepatic fibrosis in biliary tract dysplasia.In children of biliary tract dysplasia aged above 180 days,the expression of Notch2 was down-regulated with the worsening degree of hepatic fibrosis.It implied that it was correlated with a slow progression of hepatic fibrosis in biliary tract dysplasia.