中华儿科杂志
中華兒科雜誌
중화인과잡지
Chinese Journal of Pediatrics
2011年
11期
834-838
,共5页
梁斐%朱建幸%夏红萍%李慧萍%朱履昌
樑斐%硃建倖%夏紅萍%李慧萍%硃履昌
량비%주건행%하홍평%리혜평%주리창
支气管肺发育不良%血管生成素-1%氧
支氣管肺髮育不良%血管生成素-1%氧
지기관폐발육불량%혈관생성소-1%양
Bronchopulmonary dysplasia%Angiopoietin-1%Oxygen
目的 观察持续吸入85％氧气对新生大鼠肺血管发育和肺组织血管生成素1(Ang-1)表达的影响,探讨高氧肺损伤新生大鼠的肺血管发育情况及可能的发生机制.方法 将96只新生SD大鼠在生后6h内,随机分为高氧组和空气组,高氧组将大鼠置于自制密闭氧箱,FiO2=0.85.在实验3、7、14d每组各随机取16只处死,采集标本.采用HE染色进行肺组织形态学分析,放射状肺泡计数评价肺泡化程度,肺动脉钡剂造影及肺动脉密度检测评价肺血管的发育,免疫组织化学法检测肺组织Ang-1的表达,荧光定量PCR和Western blot技术检测肺组织Ang-1的mRNA和蛋白表达水平.结果 (1)新生大鼠高氧暴露14 d,肺组织的病理表现与早产儿支气管肺发育不良(BPD)的病理表现相似.(2)高氧组7 d RAC值显著低于空气组[(10.55±0.13):(11.74 ±0.19),P＜0.05],在14d时差异更显著[(12.47±0.05):( 15.03±0.16),P＜0.01].(3)肺动脉钡剂造影显示,高氧组14 d大鼠肺动脉主干变细,肺小动脉显影减少,肺动脉密度显著低于空气组[(3.55±0.09):(6.03±0.16),P＜0.05].(4)免疫组化染色显示,高氧组7d和14 d,肺组织Ang-1的表达均显著低于同时间点空气组[ (4.27±0.34):(3.10 ±0.29),P＜0.05,(5.65±0.49)∶(3.21±0.28),P＜0.01].(5)荧光定量PCR及Western blot结果显示,高氧组7d和14 d,Ang-1的mRNA水平显著低于空气组[(0.85±0.14)∶(0.44±0.21),P＜0.05,(0.87±0.24)∶(0.24±0.05),P＜0.01],Ang-1的蛋白水平也显著低于空气组[(0.88±0.31)∶(0.41 ±0.12),P＜0.05,(0.90 ±0.29):(0.21 ±0.06),P＜0.01].结论 持续吸入高浓度氧导致新生大鼠的肺血管发育障碍,Ang-1的表达下调可能参与了早产儿BPD血管发育受阻的发生机制.
目的 觀察持續吸入85％氧氣對新生大鼠肺血管髮育和肺組織血管生成素1(Ang-1)錶達的影響,探討高氧肺損傷新生大鼠的肺血管髮育情況及可能的髮生機製.方法 將96隻新生SD大鼠在生後6h內,隨機分為高氧組和空氣組,高氧組將大鼠置于自製密閉氧箱,FiO2=0.85.在實驗3、7、14d每組各隨機取16隻處死,採集標本.採用HE染色進行肺組織形態學分析,放射狀肺泡計數評價肺泡化程度,肺動脈鋇劑造影及肺動脈密度檢測評價肺血管的髮育,免疫組織化學法檢測肺組織Ang-1的錶達,熒光定量PCR和Western blot技術檢測肺組織Ang-1的mRNA和蛋白錶達水平.結果 (1)新生大鼠高氧暴露14 d,肺組織的病理錶現與早產兒支氣管肺髮育不良(BPD)的病理錶現相似.(2)高氧組7 d RAC值顯著低于空氣組[(10.55±0.13):(11.74 ±0.19),P＜0.05],在14d時差異更顯著[(12.47±0.05):( 15.03±0.16),P＜0.01].(3)肺動脈鋇劑造影顯示,高氧組14 d大鼠肺動脈主榦變細,肺小動脈顯影減少,肺動脈密度顯著低于空氣組[(3.55±0.09):(6.03±0.16),P＜0.05].(4)免疫組化染色顯示,高氧組7d和14 d,肺組織Ang-1的錶達均顯著低于同時間點空氣組[ (4.27±0.34):(3.10 ±0.29),P＜0.05,(5.65±0.49)∶(3.21±0.28),P＜0.01].(5)熒光定量PCR及Western blot結果顯示,高氧組7d和14 d,Ang-1的mRNA水平顯著低于空氣組[(0.85±0.14)∶(0.44±0.21),P＜0.05,(0.87±0.24)∶(0.24±0.05),P＜0.01],Ang-1的蛋白水平也顯著低于空氣組[(0.88±0.31)∶(0.41 ±0.12),P＜0.05,(0.90 ±0.29):(0.21 ±0.06),P＜0.01].結論 持續吸入高濃度氧導緻新生大鼠的肺血管髮育障礙,Ang-1的錶達下調可能參與瞭早產兒BPD血管髮育受阻的髮生機製.
목적 관찰지속흡입85％양기대신생대서폐혈관발육화폐조직혈관생성소1(Ang-1)표체적영향,탐토고양폐손상신생대서적폐혈관발육정황급가능적발생궤제.방법 장96지신생SD대서재생후6h내,수궤분위고양조화공기조,고양조장대서치우자제밀폐양상,FiO2=0.85.재실험3、7、14d매조각수궤취16지처사,채집표본.채용HE염색진행폐조직형태학분석,방사상폐포계수평개폐포화정도,폐동맥패제조영급폐동맥밀도검측평개폐혈관적발육,면역조직화학법검측폐조직Ang-1적표체,형광정량PCR화Western blot기술검측폐조직Ang-1적mRNA화단백표체수평.결과 (1)신생대서고양폭로14 d,폐조직적병리표현여조산인지기관폐발육불량(BPD)적병리표현상사.(2)고양조7 d RAC치현저저우공기조[(10.55±0.13):(11.74 ±0.19),P＜0.05],재14d시차이경현저[(12.47±0.05):( 15.03±0.16),P＜0.01].(3)폐동맥패제조영현시,고양조14 d대서폐동맥주간변세,폐소동맥현영감소,폐동맥밀도현저저우공기조[(3.55±0.09):(6.03±0.16),P＜0.05].(4)면역조화염색현시,고양조7d화14 d,폐조직Ang-1적표체균현저저우동시간점공기조[ (4.27±0.34):(3.10 ±0.29),P＜0.05,(5.65±0.49)∶(3.21±0.28),P＜0.01].(5)형광정량PCR급Western blot결과현시,고양조7d화14 d,Ang-1적mRNA수평현저저우공기조[(0.85±0.14)∶(0.44±0.21),P＜0.05,(0.87±0.24)∶(0.24±0.05),P＜0.01],Ang-1적단백수평야현저저우공기조[(0.88±0.31)∶(0.41 ±0.12),P＜0.05,(0.90 ±0.29):(0.21 ±0.06),P＜0.01].결론 지속흡입고농도양도치신생대서적폐혈관발육장애,Ang-1적표체하조가능삼여료조산인BPD혈관발육수조적발생궤제.
Objective To study the effects of prolonged 85％ oxygen exposure on lung vascular development and the expression of angiopoietin-1 (Ang-1) in the neonatal rat lungs.Methods Ninety-six Sprague-Dawley rat pups were randomly exposed to air (control group) and 85％ oxygen (experimental group) 6 hrs after birth.The rats were sacrificed 3,7 and 14 days after exposure and their lungs were sampled.The lung sections were stained with hematoxylin and eosin for histological evaluation and analysis of vessel volume density.Expressions of angiopoietin-1 (Ang-1) in lung tissue were measured by immunohistochemistry.Expression of Ang-1 protein and mRNA was detected by Western Blot and Real timePCR.Results After being exposed to 85％ oxygen for 14 days,lung tissues had pathological changes as "new" bronchopulmonary dysplasia (BPD).The RAC on day 7 and day 14 in experimental group decreased significantly as compared with the control group [ ( 10.55 ±0.13 ) vs.( 1 1.74 ±0.19),( 12.47 ±0.05 )vs.( 15.03 ±0.16),P ＜0.05].The X-ray showed that the diameter of lung vessel was much smaller and the vessels had less branches in experimental group compared with the control group on day 14.The vessel volume density on day 14 in experimental group decreased significantly as compared with the control group [ (3.55 ± 0.09) vs.(6.03 ± 0.16 ),P ＜ 0.05 ].Immunohistochemistry and Western blotting showed that the expressions of Ang-1 protein on day 7 and day 14 in the experimental group decreased significantly as compared with the control group [ (4.27 ± 0.34 ) vs.( 3.10 ± 0.29 ),P ＜ 0.05,( 5.65 ± 0.49 ) vs.(3.21±0.28),P＜0.01],[(0.88±0.31) vs.(0.41±0.12),P＜0.05,(0.90±0.29) vs.(0.21±0.06),P＜0.01 ].The expressions of Ang-1 mRNA on day 7 and day 14 in the experimental group also decreased significantly as compared with the control group [ (0.85 ±0.14) vs.(0.44 ±0.21 ),P ＜0.05,(0.87 ± 0.24 ) vs.(0.24 ± 0.05 ),P ＜ 0.01 ].Conclusions Prolonged exposure of high concentration of oxygen may cause impairment of lung vascular development by inhibiting expression of Ang-1 in neonatal rats,which is likely to contribute to pathogenesis of BPD.
