中华小儿外科杂志
中華小兒外科雜誌
중화소인외과잡지
CHINESE JOURNAL OF PEDIATRIC SURGERY
2014年
3期
212-217
,共6页
肖尚杰%何秋明%杨文熠%朱士博%夏慧敏
肖尚傑%何鞦明%楊文熠%硃士博%夏慧敏
초상걸%하추명%양문습%주사박%하혜민
疝,横膈%支气管肺发育不良%地塞米松
疝,橫膈%支氣管肺髮育不良%地塞米鬆
산,횡격%지기관폐발육불량%지새미송
Hernia,diaphragmatic%Bronchopulmonary dysplasia%Dexamethasone
目的 研究产前应用地塞米松(Dex)对除草醚(Nitrofen)诱导的先天性膈疝(congenital diaphragmatic hernia,CDH)大鼠模型中胎肺发育及胎肺中Wnt7b表达的影响,探讨地塞米松改善CDH肺发育不良的可能机制.方法 采用数字随机法将12只孕鼠分为3组,利用Nitrofen诱导制作CDH胎鼠模型.其中,地塞米松组(CDH+ Dex组)与CDH组在孕第9.5天给予Nitrofen,前者产前应用地塞米松治疗,后者则不予地塞米松治疗,未作任何治疗的正常胎鼠为对照组.于孕21.5d行剖腹产,解剖胎鼠,取出胎肺.采用组织学检查方法评估各组胎肺的发育情况.采用实时荧光定量PCR方法检测各组胎肺中Wnt7b mRNA的表达情况.采用免疫组织化学方法检测各组胎肺中Wnt7b蛋白的表达情况.结果 成功构建CDH胎鼠模型.对照组胎肺发育较为成熟,CDH组胎肺发育明显滞后,CDH+ Dex组胎肺发育有所改善.对照组、CDH组和CDH+ Dex组的平均肺泡面积分别为(1 443.37±285.94)μm2、(332.83±72.19)μm2和(929.36±110.31)μm2;平均肺泡间隔厚度分别为(6.17±1.54)μm、(9.72±2.18)μm和(7.01±1.32)μm;肺血管数/HP分别为(7.20±0.91)个、(3.68±1.03)个和(6.84±0.78)个;管腔面积占血管总面积比分别为(38,74±4.94)％、(15.76±4.87)％和(37.12±7.03)％.与对照组相比,CDH组胎肺平均肺泡面积减少,平均肺泡间隔增厚,每高倍视野肺血管数减少,血管管腔狭窄,组间差异均有统计学意义(均P＜0.01);与CDH组相比,CDH+ Dex组胎肺平均肺泡面积增大,平均肺泡间隔厚度减小,每高倍视野肺血管数增加,血管管腔增大,组间差异亦有统计学意义(均P＜0.01).以21.5d胎龄的对照组胎肺作为对照样本,CDH组和CDH+ Dex组胎肺Wnt7b mRNA的相对表达量分别为1.58±0.19和0.72±0.15.CDH组胎肺的Wnt7b mRNA表达量比对照组升高,差异有统计学意义(P＜0.01);CDH+ Dex组胎肺的Wnt7bmRNA表达量较CDH组明显降低,差异有统计学意义(P＜0.01).Wnt7b免疫阳性细胞主要分布在支气管和细支气管上皮.结论 产前应用地塞米松可明显改善CDH胎鼠肺发育不良,同时下调胎肺中Wnt7b mRNA的表达,推测地塞米松可能通过调节Wnt7b的表达来改善CDH肺发育不良.
目的 研究產前應用地塞米鬆(Dex)對除草醚(Nitrofen)誘導的先天性膈疝(congenital diaphragmatic hernia,CDH)大鼠模型中胎肺髮育及胎肺中Wnt7b錶達的影響,探討地塞米鬆改善CDH肺髮育不良的可能機製.方法 採用數字隨機法將12隻孕鼠分為3組,利用Nitrofen誘導製作CDH胎鼠模型.其中,地塞米鬆組(CDH+ Dex組)與CDH組在孕第9.5天給予Nitrofen,前者產前應用地塞米鬆治療,後者則不予地塞米鬆治療,未作任何治療的正常胎鼠為對照組.于孕21.5d行剖腹產,解剖胎鼠,取齣胎肺.採用組織學檢查方法評估各組胎肺的髮育情況.採用實時熒光定量PCR方法檢測各組胎肺中Wnt7b mRNA的錶達情況.採用免疫組織化學方法檢測各組胎肺中Wnt7b蛋白的錶達情況.結果 成功構建CDH胎鼠模型.對照組胎肺髮育較為成熟,CDH組胎肺髮育明顯滯後,CDH+ Dex組胎肺髮育有所改善.對照組、CDH組和CDH+ Dex組的平均肺泡麵積分彆為(1 443.37±285.94)μm2、(332.83±72.19)μm2和(929.36±110.31)μm2;平均肺泡間隔厚度分彆為(6.17±1.54)μm、(9.72±2.18)μm和(7.01±1.32)μm;肺血管數/HP分彆為(7.20±0.91)箇、(3.68±1.03)箇和(6.84±0.78)箇;管腔麵積佔血管總麵積比分彆為(38,74±4.94)％、(15.76±4.87)％和(37.12±7.03)％.與對照組相比,CDH組胎肺平均肺泡麵積減少,平均肺泡間隔增厚,每高倍視野肺血管數減少,血管管腔狹窄,組間差異均有統計學意義(均P＜0.01);與CDH組相比,CDH+ Dex組胎肺平均肺泡麵積增大,平均肺泡間隔厚度減小,每高倍視野肺血管數增加,血管管腔增大,組間差異亦有統計學意義(均P＜0.01).以21.5d胎齡的對照組胎肺作為對照樣本,CDH組和CDH+ Dex組胎肺Wnt7b mRNA的相對錶達量分彆為1.58±0.19和0.72±0.15.CDH組胎肺的Wnt7b mRNA錶達量比對照組升高,差異有統計學意義(P＜0.01);CDH+ Dex組胎肺的Wnt7bmRNA錶達量較CDH組明顯降低,差異有統計學意義(P＜0.01).Wnt7b免疫暘性細胞主要分佈在支氣管和細支氣管上皮.結論 產前應用地塞米鬆可明顯改善CDH胎鼠肺髮育不良,同時下調胎肺中Wnt7b mRNA的錶達,推測地塞米鬆可能通過調節Wnt7b的錶達來改善CDH肺髮育不良.
목적 연구산전응용지새미송(Dex)대제초미(Nitrofen)유도적선천성격산(congenital diaphragmatic hernia,CDH)대서모형중태폐발육급태폐중Wnt7b표체적영향,탐토지새미송개선CDH폐발육불량적가능궤제.방법 채용수자수궤법장12지잉서분위3조,이용Nitrofen유도제작CDH태서모형.기중,지새미송조(CDH+ Dex조)여CDH조재잉제9.5천급여Nitrofen,전자산전응용지새미송치료,후자칙불여지새미송치료,미작임하치료적정상태서위대조조.우잉21.5d행부복산,해부태서,취출태폐.채용조직학검사방법평고각조태폐적발육정황.채용실시형광정량PCR방법검측각조태폐중Wnt7b mRNA적표체정황.채용면역조직화학방법검측각조태폐중Wnt7b단백적표체정황.결과 성공구건CDH태서모형.대조조태폐발육교위성숙,CDH조태폐발육명현체후,CDH+ Dex조태폐발육유소개선.대조조、CDH조화CDH+ Dex조적평균폐포면적분별위(1 443.37±285.94)μm2、(332.83±72.19)μm2화(929.36±110.31)μm2;평균폐포간격후도분별위(6.17±1.54)μm、(9.72±2.18)μm화(7.01±1.32)μm;폐혈관수/HP분별위(7.20±0.91)개、(3.68±1.03)개화(6.84±0.78)개;관강면적점혈관총면적비분별위(38,74±4.94)％、(15.76±4.87)％화(37.12±7.03)％.여대조조상비,CDH조태폐평균폐포면적감소,평균폐포간격증후,매고배시야폐혈관수감소,혈관관강협착,조간차이균유통계학의의(균P＜0.01);여CDH조상비,CDH+ Dex조태폐평균폐포면적증대,평균폐포간격후도감소,매고배시야폐혈관수증가,혈관관강증대,조간차이역유통계학의의(균P＜0.01).이21.5d태령적대조조태폐작위대조양본,CDH조화CDH+ Dex조태폐Wnt7b mRNA적상대표체량분별위1.58±0.19화0.72±0.15.CDH조태폐적Wnt7b mRNA표체량비대조조승고,차이유통계학의의(P＜0.01);CDH+ Dex조태폐적Wnt7bmRNA표체량교CDH조명현강저,차이유통계학의의(P＜0.01).Wnt7b면역양성세포주요분포재지기관화세지기관상피.결론 산전응용지새미송가명현개선CDH태서폐발육불량,동시하조태폐중Wnt7b mRNA적표체,추측지새미송가능통과조절Wnt7b적표체래개선CDH폐발육불량.
Objective To explore the effects of antenatal administration of dexamethasone (Dex) on fetal lung development and the expression of Wnt7b in rat model of nitrofen-induced congenital diaphragmatic hernia (CDH) and elucidate the possible mechanism of prenatal dexamethasone in improving pulmonary hypoplasia associated with CDH.Methods A total of 12 pregnant rats were randomly divided into CDH + Dex,CDH and control groups.CDH was induced in pregnant rats after a dose of 100 mg nitrofen at Day 9.5 of gestation (term,22 days).Dex (0.4 mg/kg) was given by an intraperitoneal injection at Days 18.5 and 19.5 of gestation.Cesarean section was performed at Day 21 of gestation.The fetuses were divided into 3 group of A,control; B,nitrofeninduced CDH; C,nitrofen-induced CDH with antenatal Dex treatment.The histological structures of fetal rat lungs were observed microscopically.The parameters of area of alveolar space,thickness of alveolar septum,number of pulmonary artery and diameter of pulmonary artery were compared to examine the lung development.The expression of Wnt7b mRNA in fetal lungs was detected by realtime quantitative polymerase chain reaction (PCR).And the expression of Wnt7b protein was detected by immunohistochemistry.Results Histological detection showed that prenatal Dex dosing improved lung development in CDH rats.The alveolar space of control,CDH and CDH + Dex groups was (1 443.37 ± 285.94),(332.83 ± 72.19) and (929.36 ± 110.31) μm2 respectively,the thickness of alveolar septum (6.17 ± 1.54),(9.72 ± 2.18)and (7.01 ± 1.32)μm,the number of pulmonary artery (7.20 ± 0.91),(3.68 ± 1.03) and (6.84 ± 0.78) and diameter of pulmonary artery (38.74 ± 4.94) ％,(15.76±4.87)％ and (37.12 ± 7.03)％.At Day 21,Wnt7b relative mRNA expression in CDH and CDH + Dex groups was 1.58 ± 0.19 and 0.72 ± 0.15 compared to control group.The Wnt7b expression in CDH group was significantly higher than that in control group.And it decreased significantly after prenatal Dex.Immunohistochemistry showed that Wnt7b protein was expressed predominantly in airway epithelium.Conclusions Prenatal Dex administration improves pulmonary hypoplasia associated with CDH and down-regulates the expression of Wnt7b rnRNA in fetal lung of CDH rats.It is hypothesized that prenatal Dex may improve pulmonary hypoplasia in CDH by regulating the expression of Wnt7b.
