中华小儿外科杂志
中華小兒外科雜誌
중화소인외과잡지
Chinese Journal of Pediatric Surgery
2015年
9期
666-670
,共5页
何秋明%肖尚杰%刘芬%朱士博%田松%钟微%夏慧敏
何鞦明%肖尚傑%劉芬%硃士博%田鬆%鐘微%夏慧敏
하추명%초상걸%류분%주사박%전송%종미%하혜민
疝,横膈%肺%维生素A
疝,橫膈%肺%維生素A
산,횡격%폐%유생소A
Hernia,diaphragmatic%Lung%Vitamin A
目的 产前应用维生素A(VitA)对除草醚(Nitrofen)诱导的先天性膈疝(congenital diaphragmatic 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1 ml橄榄油的Nitrofen 100 mg/只;对照组,于孕第9.5天仅注入橄榄油1 ml/只;维生素A产前干预组(CDH+ VitA组),于孕第9.5天注入Ni-trofen 100 mg/只后,再于孕第10.5天经胃管注入VitA 15 000 IU/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(Western blot,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)μm2、(1 443.37±285.94) μm2和(907.07±54.78) μm2,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Hoxa5 mRNA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表达水平较CDH组明显降低,差异有统计学意义(P＜0.05).结论 产前应用VitA可明显改善CDH模型胎肺发育不良,并下调胎肺中Hoxa5的表达水平,推测调控Hoxa5的表达可能是VitA改善CDH肺发育不良的作用机制之一.
目的 產前應用維生素A(VitA)對除草醚(Nitrofen)誘導的先天性膈疝(congenital diaphragmatic hernia,CDH)大鼠模型中胎肺晚期髮育及胎肺中Hoxa5錶達的影響,探討VitA改善CDH肺髮育不良的可能作用機製.方法 12隻孕鼠按隨機數字錶法隨機分為Nitrofen誘導CDH肺髮育不良組(CDH組),于大鼠孕第9.5天經胃管註入溶于1 ml橄欖油的Nitrofen 100 mg/隻;對照組,于孕第9.5天僅註入橄欖油1 ml/隻;維生素A產前榦預組(CDH+ VitA組),于孕第9.5天註入Ni-trofen 100 mg/隻後,再于孕第10.5天經胃管註入VitA 15 000 IU/隻.各組均于孕21.5天行剖腹產,取胎肺樣本;組織學檢查方法分析各組胎肺的髮育情況,實時熒光定量PCR(QPCR)、免疫蛋白印跡(Western blot,WB)方法檢測各組胎肺中Hoxa5錶達水平的變化.結果 CDH組膈疝髮生率為50.0％ (24/48),CDH+ VitA組膈疝髮生率明顯下降為28.3％ (13/46),差異有統計學意義(P＜0.05).組織學分析結果顯示,CDH組、對照組、CDH+ VitA組的平均肺泡麵積分彆為(332.83±72.19)μm2、(1 443.37±285.94) μm2和(907.07±54.78) μm2,平均肺泡間隔厚度分彆為(9.72±2.18)μm、(6.17±1.54) μm和(7.26±1.52) μm,肺血管數/HP分彆為(3.68±1.03)箇、(7.20±0.91)箇和(6.24±0.88)箇,管腔麵積佔血管總麵積比分彆(15.76±4.87)％、(38.74±4.94)％和(30.18±7.03)％.與CDH組比較,CDH+ VitA組胎肺的平均肺泡麵積增大、平均肺泡間隔厚度減小、肺血管數/HP增加、管腔增大,且差異均有統計學意義(均P＜0.05),肺髮育不良明顯改善.以對照組胎肺樣本作為參照,CDH組、CDH+ VitA組的Hoxa5 mRNA相對錶達量分彆為1.76±0.23、0.92±0.16;Hoxa5蛋白相對錶達量分彆為2.95±0.12、1.36±0.07.CDH+ VitA組胎肺的Hoxa5錶達水平較CDH組明顯降低,差異有統計學意義(P＜0.05).結論 產前應用VitA可明顯改善CDH模型胎肺髮育不良,併下調胎肺中Hoxa5的錶達水平,推測調控Hoxa5的錶達可能是VitA改善CDH肺髮育不良的作用機製之一.
목적 산전응용유생소A(VitA)대제초미(Nitrofen)유도적선천성격산(congenital diaphragmatic hernia,CDH)대서모형중태폐만기발육급태폐중Hoxa5표체적영향,탐토VitA개선CDH폐발육불량적가능작용궤제.방법 12지잉서안수궤수자표법수궤분위Nitrofen유도CDH폐발육불량조(CDH조),우대서잉제9.5천경위관주입용우1 ml감람유적Nitrofen 100 mg/지;대조조,우잉제9.5천부주입감람유1 ml/지;유생소A산전간예조(CDH+ VitA조),우잉제9.5천주입Ni-trofen 100 mg/지후,재우잉제10.5천경위관주입VitA 15 000 IU/지.각조균우잉21.5천행부복산,취태폐양본;조직학검사방법분석각조태폐적발육정황,실시형광정량PCR(QPCR)、면역단백인적(Western blot,WB)방법검측각조태폐중Hoxa5표체수평적변화.결과 CDH조격산발생솔위50.0％ (24/48),CDH+ VitA조격산발생솔명현하강위28.3％ (13/46),차이유통계학의의(P＜0.05).조직학분석결과현시,CDH조、대조조、CDH+ VitA조적평균폐포면적분별위(332.83±72.19)μm2、(1 443.37±285.94) μm2화(907.07±54.78) μm2,평균폐포간격후도분별위(9.72±2.18)μm、(6.17±1.54) μm화(7.26±1.52) μm,폐혈관수/HP분별위(3.68±1.03)개、(7.20±0.91)개화(6.24±0.88)개,관강면적점혈관총면적비분별(15.76±4.87)％、(38.74±4.94)％화(30.18±7.03)％.여CDH조비교,CDH+ VitA조태폐적평균폐포면적증대、평균폐포간격후도감소、폐혈관수/HP증가、관강증대,차차이균유통계학의의(균P＜0.05),폐발육불량명현개선.이대조조태폐양본작위삼조,CDH조、CDH+ VitA조적Hoxa5 mRNA상대표체량분별위1.76±0.23、0.92±0.16;Hoxa5단백상대표체량분별위2.95±0.12、1.36±0.07.CDH+ VitA조태폐적Hoxa5표체수평교CDH조명현강저,차이유통계학의의(P＜0.05).결론 산전응용VitA가명현개선CDH모형태폐발육불량,병하조태폐중Hoxa5적표체수평,추측조공Hoxa5적표체가능시VitA개선CDH폐발육불량적작용궤제지일.
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 E10.5.Pregnant rats without nitrofen were selected as control group.Fetal lungs were collected at E21.5.Histological pulmonary development was analyzed by hematoxylin & 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) μm2;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 nitrofeninduced CDH and down-regulates the expression of Hoxa5.And the improvement of pulmonary hypoplasia may be regulated by the expression of Hoxa5.
