CAJ | 학술논문

目的研究Wnt7b在Nitrofen诱导的先天性膈疝(congenital diaphragmatic hernia,CDH)胎肺中的表达特点,探讨其在CDH肺发育不良发生机制中的可能作用.方法采用实时荧光定量PCR方法检测Wnt7b基因在妊娠E17.5、E19.5、E21.5的Nitrofen诱导CDH大鼠模型胎肺及正常对照大鼠胎肺中的相对表达量,并用免疫组化方法检测Wnt7b蛋白在胎肺中的表达.结果正常对照组和CDH组胎肺中Wnt7b mRNA的表达水平均随着胎龄的增加呈下降趋势,其中E17.5胎龄CDH组胎肺中Wnt7b mRNA的表达水平与对照组相比,差异无统计学意义;而E19.5、E21.5胎龄CDH组胎肺中Wnt7b mRNA的表达水平高于对照组,差异有统计学意义(P＜0.05).对E21.5胎龄对照组和CDH组的标本进行免疫组化检测,结果显示Wnt7b免疫阳性细胞主要分布在支气管和细支气管上皮,且CDH组Wnt7b免疫阳性程度(以平均光密度值表示)较正常对照组升高,有统计学差异.结论 Wnt7b mRNA表达量随大鼠胎肺逐渐发育成熟而下降,提示其在胎肺中的表达量与胎肺发育的成熟度有关.CDH组中E19.5、E21.5胎龄时Wnt7b mRNA的表达水平明显高于对照组,是CDH孕晚期胎肺发育滞后的分子基础之一,提示Wnt7b在CDH孕晚期胎肺中表达增高可能参与了肺发育不良.Wnt7b蛋白在肺发育中特异性定位于气道上皮,提示信号町能在肺上皮与间质细胞之间有重要联系,协同促进上皮和间质的发育.
목적 연구Wnt7b재Nitrofen유도적선천성격산(congenital diaphragmatic hernia,CDH)태폐중적표체특점,탐토기재CDH폐발육불량발생궤제중적가능작용.방법 채용실시형광정량PCR방법검측Wnt7b기인재임신E17.5、E19.5、E21.5적Nitrofen유도CDH대서모형태폐급정상대조대서태폐중적상대표체량,병용면역조화방법검측Wnt7b단백재태폐중적표체.결과 정상대조조화CDH조태폐중Wnt7b mRNA적표체수평균수착태령적증가정하강추세,기중E17.5태령CDH조태폐중Wnt7b mRNA적표체수평여대조조상비,차이무통계학의의;이E19.5、E21.5태령CDH조태폐중Wnt7b mRNA적표체수평고우대조조,차이유통계학의의(P＜0.05).대E21.5태령대조조화CDH조적표본진행면역조화검측,결과현시Wnt7b면역양성세포주요분포재지기관화세지기관상피,차CDH조Wnt7b면역양성정도(이평균광밀도치표시)교정상대조조승고,유통계학차이.결론 Wnt7b mRNA표체량수대서태폐축점발육성숙이하강,제시기재태폐중적표체량여태폐발육적성숙도유관.CDH조중E19.5、E21.5태령시Wnt7b mRNA적표체수평명현고우대조조,시CDH잉만기태폐발육체후적분자기출지일,제시Wnt7b재CDH잉만기태폐중표체증고가능삼여료폐발육불량.Wnt7b단백재폐발육중특이성정위우기도상피,제시신호정능재폐상피여간질세포지간유중요련계,협동촉진상피화간질적발육.
Objective To investigate the Wnt7b expression in fetal lungs of rat models of congenital diaphragmatic hernia(CDH) induced by nitrofen and to investigate the role of Wnt7b in the formation of pulmonary dysplasia in the process of CDH.Methods The expression of Wnt7b mRNA were detected in different developmental stages (E17.5,E19.5 and E21.5) in fetal lungs of rat model of CDH and in lungs of normal rats by Real-time Quantitative RT-PCR The expression of Wnt7b protein in rat fetal lungs were detected by immunohistochemistry.Results The relative quantities of Wnt7b mRNA in the fetal lungs decreased during E17.5,E19.5 and E21.5 both in normal group and CDH group.The Wnt7b expression was significantly higher in lungs at E19.5 and E21.5 in lungs of nitrofen-treated rats than that in normal lungs,while at E17.5,no significant difference were noted between both groups.Immunohistochemical staining showed that Wnt7b was expressed predominantly in the airway epithelia.The immunological activity (shown by the average optical density) in CDH group was higher than that in the control group.Conclusions The Wnt7b mRNA expression decreases gradually with development of fetal lungs,which may indicate that Wnt7b mRNA probably associates with the maturity of fetallungs.The relatively increased level of Wnt7b mRNA in CDH group at the late gesration may be the molecular basis of pulmonary dysplasia in rat models of CDH.The exclusive expression of Wnt7b in the airway epithelia indicates that Wnt7b signaling pathway may be important between epithelia and mesenchymal,which suggests that Wnt7b can improve the concordant development of epithelia and mesenchymal.