CAJ | 학술논문

目的研究TNF-α与IL-1β在除草醚(Nitrofen)诱导的先天性膈疝(CDH)大鼠模型中的表达及意义.方法分别用免疫组织化学法与ELISA法检测TNF-α与IL-1β蛋白在在Nitrofen诱导CDH大鼠模型的胎肺与羊水中的表达；及采用实时荧光定量PCR法检测TNF-α与IL-1β基因在上述CDH大鼠模型胎肺及正常对照大鼠胎肺中的相对表达量(妊娠第16.5、18.5和21天时).结果免疫组织化学检测结果显示,Nitrofen组各时期胎肺中TNF-α表达均强于对照组(P＜0.05),而IL-1β仅在孕21 d表达强于对照组(P＜0.05).对孕21 d胎鼠的羊水标本行ELISA检测,结果显示Nitrofen组TNF-α与IL-1β含量在孕21 d时较对照组高,差异有统计学意义(P＜0.05).正常对照组胎肺中TNF-α与IL-1βmRNA表达水平在上述3个胎龄中以第16.5天表达量最高,之后随着胎龄增加呈下降趋势；Nitrofen组TNF-α与IL-1βmRNA表达趋势与正常胎肺相似,但表达量均显著高于同龄对照组,差异有统计学意义(P＜0.05).结论 Nitro fen可干扰CDH胎鼠肺组织中TNF-α与IL-1β的表达,并能引起羊水中TNF-α与IL-1β含量出现显著变化,这可能是其导致CDH胎肺发育异常的原理机制之一.
목적 연구TNF-α여IL-1β재제초미(Nitrofen)유도적선천성격산(CDH)대서모형중적표체급의의.방법 분별용면역조직화학법여ELISA법검측TNF-α여IL-1β단백재재Nitrofen유도CDH대서모형적태폐여양수중적표체；급채용실시형광정량PCR법검측TNF-α여IL-1β기인재상술CDH대서모형태폐급정상대조대서태폐중적상대표체량(임신제16.5、18.5화21천시).결과 면역조직화학검측결과현시,Nitrofen조각시기태폐중TNF-α표체균강우대조조(P＜0.05),이IL-1β부재잉21 d표체강우대조조(P＜0.05).대잉21 d태서적양수표본행ELISA검측,결과현시Nitrofen조TNF-α여IL-1β함량재잉21 d시교대조조고,차이유통계학의의(P＜0.05).정상대조조태폐중TNF-α여IL-1βmRNA표체수평재상술3개태령중이제16.5천표체량최고,지후수착태령증가정하강추세；Nitrofen조TNF-α여IL-1βmRNA표체추세여정상태폐상사,단표체량균현저고우동령대조조,차이유통계학의의(P＜0.05).결론 Nitro fen가간우CDH태서폐조직중TNF-α여IL-1β적표체,병능인기양수중TNF-α여IL-1β함량출현현저변화,저가능시기도치CDH태폐발육이상적원리궤제지일.
Objective The aim of this study was to investigate the dynamic expression of TNF-α and IL-1β in nitrofen-induced congenital diaphragmatic hernia (CDH) rat model.Methods Pregnant female Sprague-Dawley rats were randomly divided into 2 groups including control and nitrofen group.The rats were exposed to either olive oil or 100 mg of nitrofen at day 9.5 of gestation (D 9.5).Fetal lung tissues were harvested at D 16.5,D 18.5 and D 21 respectively.Real-time quantitative PCR and immunohistochemistry staining were performed to evaluate the level of TNF-α and IL-1β in lung tissues.Amniotic fluid was collected at D 21 and TNF-α and IL-1β concentrations were measured via ELISA assay.Results The expression of TNF-α protein in fetal lungs (at each time point) and amniotic fluid (at D 21) in nitrofen group was higher than that in control (P＜0.05),while no significant differences were noted for the expression of IL-1β protein between both groups at D 16.5 and 18.5 (P＞0.05).The TNF-α and IL-1β gene expression in lung tissues decreased at D 16.5,D 18.5 and D 21 in both groups.The TNF-α and IL-1β gene expression levels were found significantly higher in nitrofen-treated rats than control at each time point (P＜0.05).Conclusions The expression perturbation of TNF-α and IL-1β in nitrofen-induced CDH rat model was caused by nitrofen,which results in excessive production of the pro-inflammatory cytokine TNF-α and IL-1β in CDH rat model.It may be one of the feasible mechanisms that lead to pulmonary hypoplasia and increased neonatal mortality.