摘要
目的产前应用维生素A（vitA）对除草醚（Nitrofen）诱导的先天性膈疝（congenitaldi—aphragmatic hernia，CDH）大鼠模型中胎肺晚期发育及胎肺中Hoxa5表达的影响，探讨VitA改善CDH肺发育不良的可能作用机制。方法12只孕鼠按随机数字表法随机分为Nitrofen诱导CDH肺发育不良组（CDH组），于大鼠孕第9．5天经胃管注入溶于1ml橄榄油的Nitrofen100mg／只；对照组，于孕第9．5天仅注入橄榄油1ml／只；维生素A产前干预组（CDH＋VitA组），于孕第9．5天注入Ni—trofen 100mg／只后，再于孕第10．5天经胃管注入VitA15000IU／只。各组均于孕21．5天行剖腹产，取胎肺样本；组织学检查方法分析各组胎肺的发育情况，实时荧光定量PCR（QPCR）、免疫蛋白印迹（Westernblot，WB）方法检测各组胎肺中Hoxa5表达水平的变化。结果CDH组膈疝发生率为50．0％（24／48），CDH＋VitA组膈疝发生率明显下降为28．3％（13／46），差异有统计学意义（P〈0．05）。组织学分析结果显示，CDH组、对照组、CDH＋VitA组的平均肺泡面积分别为（332．83±72．19）μm^2、（1443．37±285．94）μm^2和（907．07±54．78）μm^2，平均肺泡间隔厚度分别为（9．72±2．18）μm、（6．17±1．54）μm和（7．26±1．52）μm，肺血管数／HP分别为（3．68±1．03）个、（7．20±0．91）个和（6．24±0．88）个，管腔面积占血管总面积比分别（15．76±4．87）％、（38．74±4．94）％和（30．18±7．03）％。与CDH组比较，CDH＋VitA组胎肺的平均肺泡面积增大、平均肺泡间隔厚度减小、肺血管数／HP增加、管腔增大，且差异均有统计学意义（均P〈0．05），肺发育不良明显改善。以对照组胎肺样本作为参照，CDH组、CDH＋VitA组的Hoxa5mRNA相对表达量分别为1．76±0．23、0．92±0．16；Hoxa5蛋白相对表达量分别为2．95±0．12、1．36±0．07。CDH＋VitA组胎肺的Hoxa5表达水�
Objective To explore the effects of prenatal administration of vitamin A （VitA） on late fetal lung development and expression of Hoxa5 and elucidate the possible mechanism of pulmonary maturation in a rat model of congenital diaphragmatic hernia （CDH）. Methods A total of 12 pregnant rats were randomly divided into CDH, CDH ± VitA and control groups. CDH was induced in pregnant rats after administration of 100 mg nitrofen at E9. 5 （day 9. 5 of gestation, term in 22 days）. Nitrofen-induced CDH ± VitA group received VitA （15 000 IU dissolved in 1 ml oil） i.g. at El0. 5. Pregnant rats without nitrofen were selected as control group. Fetal lungs were collected at E21.5. Histological pulmonary development was analyzed by hematoxylin I~ eosin staining. The expressions of Hoxa5 mRNA and protein were detected by real-time quantitative polymerase chain reaction （qRT-PCR） and Western blot respectively. Results The rate of CDH decreased markedly after prenatal VitA （CDH group 50. 0% vs CDH ± VitA group 28. 3%）. The areas of alveolar space of CDH, control and CDH ± VitA groups were （332. 83 ± 72. 19）, （1 443. 37 ± 285.94）and （907. 07 ± 54. 78） μm^2; thickness of alveolar septum （9. 72 ± 2. 18）, （6. 17 ± 1.54） and （7. 26 ± 1.52）μm; number of pulmonary artery 3. 68 ± 1.03, 7. 20 ± 0. 91 and 6. 24 ± 0. 88% luminal area to total transectional area （15.76 ± 4.87）%, （38.74 ± 4.94）% and （30.18 ± 7.03）% respectively. Histological finding showed that antenatal administration of VitA promoted lung maturity in CDH ± VitA group. The expression of Hoxa5 significantly decreased in CDH ± VitA group versus CDH group. Conclusions Prenatal administration of VitA promotes pulmonary maturation in nitrofen- induced CDH and down-regulates the expression of Hoxa5. And the improvement of pulmonary hypoplasia may be regulated by the expression of HoxaS.
出处
《中华小儿外科杂志》
CSCD
2015年第9期666-670,共5页
Chinese Journal of Pediatric Surgery
基金
基金项目：广东省自然科学基金（S2013020012738）
关键词
疝
横膈
肺
维生素A
Hernia, diaphragmatic
Lung
Vitamin A
