支气管肺发育不良新生大鼠肺组织Occludin的表达及其对肺上皮通透性的影响  

Expression of Occludin in lung tissues of newborn rats with bronchopulmonary dysplasia and its influence on pulmonary epithelial permeability

作　　者：徐姝燕[1] 富建华[1] 由凯[1] 薛辛东[1] 

机构地区：[1]中国医科大学附属盛京医院儿科,沈阳110004

出　　处：《中华实用儿科临床杂志》2013年第16期1257-1260,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（81170605;30872781）

摘　　要：目的研究高体积分数氧（高氧）致新生大鼠支气管肺发育不良（BPD）早期肺组织Occludin蛋白及mRNA的动态变化，以及其对肺上皮通透性的影响。方法新生Wistar大鼠160只，依据吸氧体积分数（FiO2）随机分为高氧组（FiO2=900mL／L）和空气组（Fi02=210mL／L），每组80只。分别于建立模型后1、3、5、7d处死大鼠后取肺组织。检测支气管肺泡灌洗液（BALF）与血清异硫氰酸荧光素标记的右旋糖酐（FIM）比值；采用免疫荧光染色及Western blot技术观察Occludin蛋白的分布和表达；采用反转录-PCR技术检测Occlu-din mRNA的表达。结果空气组与高氧组1 d BALF与血清FD4比值比较差异无统计学意义（1．533±O．122比1．575±0．140，P〉0．05）；与空气组比较，高氧组3dBALF与血清FIM比值有所增加（1．365±0．159比1．615±0．196，P〈0．05），5d、7d显著增力口（1．245±0．152比3．211±0．799；1．178±0．594比5．150±0．967，P均〈0．01）。免疫荧光结果显示，空气组7d肺泡上皮细胞边缘Occludin蛋白呈现连续线性分布，高氧组7d表达不连续，荧光强度较弱；空气组与高氧组1dOccludinmRNA表达比较差异无统计学意义（2．15±0．33比2．23±0．39，P〉0．05）；与空气组比较，高氧组3d、5dOccludin mRNA减少（2．46±0．27比2．00±0．19，2．62±0．28比2．15±0．20，P均〈0．05），7d减少最明显（3．08±0．43比2．01±0．34，P〈0．01）；1d、3d空气组与高氧组Occludin蛋白表达比较差异无统计学意义（1．00±0．05比1．05±0．03，1．24±0．06比1．17±0．04，P均〉0．05），与空气组比较，高氧组5d减少，差异有统计学意义（1．03±0．04比0．93±O．05，P〈0．05），7d减少最明显（0．96±0．14比0．65±O．07，P〈0．01）；高氧组Occludin蛋白的表达与肺上皮通透性呈明显负相关（r=-0．755，P〈0．01）。结论肺组织Occludin表达下调，可能导致Objective To explore the dynamic changes of Occludin gene and protein levels in lung tissues of newborn rats with hyperoxia-induced bronchopulmonary dysplasia （BPD） in the early phase and its effect on pulmonary epithelial permeability. Methods One hundred and sixty newborn Wistar rats were randomly assigned to hyperoxia group （900 mL/L oxygen） and normoxia group （ 210 mL/L oxygen） according to different oxygen concentrations, 80 rats in each group. Rats were sacrificed and lung tissues were removed on 1,3,5,7 d after treatment. Bronchoalveolar lavage fluid（BALF） ：serum FD4 ratio was detected;location and expression of Occludin were examined by immunofluo- rescence staining and Western blot; messenger RNA （mRNA） was studied by reverse transcription-PCR. Results There was no obvious difference in the BALF and serum FD4 ratio （1. 533 ± 0. 122 vs 1. 575 ± 0. 140, P 〉 0.05 ） between the hyperoxia group and the normoxia group on the first day. After 3 days of hyperoxia exposure ,the ratio of FD4 between BALF and serum was significantly higher than that in the normoxia group （ 1. 365 ± O. 159 vs 1. 615 ± 0. 196, P 〈 0.05 ）. And after 5 or 7 days of hyperoxia exposure, the ratio of FD4 between BALF and serum was dramatically increased（ 1. 245 ± 0. 152 vs 3. 211 ±0. 799,1. 178 ± 0. 594 vs 5.15 ± 0. 967, all P 〈 0.01 ）. On the 7 day, immunofluorescence staining showed Occludin distribllted in a consecutive line along lung epithelial cell membrane in the normoxia group ,while in the hyperoxia group Occludin was distributed in a discontinuous line and lacking intensity. There was no obvious difference in Occludin mRNA level between the hyperoxia group and normoxia group on the first day（2.15 ± 0.33 vs 2.23±0.39,P 〉0.05）. Compared to the normoxia group,the decrease in Occludin mRNA level was statistically significant after 3 or 5 days of hyperoxia exposure （ 2.46 ± 0.27 vs 2. O0 ± 0.19,2.62 ± 0.28 vs 2.15 ± 0.20, all P 〈 0.05 ）, and after 7 days of hyperoxia e

关 键 词：OCCLUDIN 肺上皮通透性 支气管肺发育不良 高体积分数氧 新生鼠 

分 类 号：R722.1[医药卫生—儿科]