摘要
目的探讨前B细胞集落增强因子(PBEF)在高氧致新生大鼠支气管肺发育不良中的表达及意义。方法 48只足月新生SD大鼠在出生12 h内根据吸氧浓度的不同分为实验组和对照组各24只。实验组持续暴露于800 ml/L的氧气中,对照组置于室内空气中。两组分别于实验开始后3 d、7 d、14 d各随机取8只新生鼠麻醉后处死,取材。HE染色光镜下观察其肺组织形态结构改变,并检测肺组织放射状肺泡计数和呼吸膜厚度。免疫组织化学方法检测PBEF的表达定位。实时荧光定量-PCR技术检测PBEF mRNA,Western blot技术和酶联免疫吸附实验分别检测肺组织和支气管肺泡灌洗液中PBEF的蛋白表达。结果 (1)HE染色观察肺组织形态学结构显示,实验组随着日龄的增加肺泡逐渐融合、肺泡正常结构减少或消失;7 d、14 d实验组放射状肺泡计数较对照组明显减少(P<0.05),呼吸膜厚度较对照组显著增大(P<0.01)。(2)免疫组化结果显示,PBEF主要表达于支气管上皮细胞、肺泡上皮细胞和肺血管内皮细胞中,实验组各时间点肺组织PBEF阳性细胞累积光密度值均显著高于对照组(P<0.05)。(3)实时荧光定量-PCR技术和Western blot结果显示,实验组3 d、7 d肺组织PBEF mRNA显著高于同时间点对照组;实验组各时间点肺组织PBEF蛋白的表达均显著高于对照组(P<0.05)。结论 PBEF参与了支气管肺发育不良的发生发展。
Objective To explore the expression and significance of Pre-B cell colony enhancing factor( PBEF) in the lungs of newborn rats with hyperoxia-induced bronchopulmonary dysplasia( BPD). Methods Fortyeight full-term newborn Sprague- Dawley rats were randomly divided into the experimental group( hyperoxia,n = 24)and the control group( room air,n = 24) according to the fraction of inspiratory( Fi O2) within 12 hours after birth.The rats of experimental group stayed in the space of 800 ml·L- 1oxygen,while the rats of the control group were exposed to room air. On the 3th,7th,14 th day of the experiment,8 newborn rats in the experimental group and other 8rats in the control group were randomly sacrificed after anesthesia. The sections of lungs were stained with HE method to assess lung histology changes and the lung radical alveolar counts( RAC) were examined. The location and semiquantitative analysis of PBEF in lungs were detected by immunohistochemical method. The PBEF mRNA expression levels were detected by reverse transcription polymerase chain reaction( RT-PCR). The protein expression levels of PBEF in lungs and bronchoalveolar lavage fluid( BALF) were determined by Western blot and ELISA respectively.Results( 1) Stained by HE,the lungs the of experimental group showed destroy of alveoli,alveoli fusion and increased septal wall thickness. The radical alveolar counts were significantly lower in the experimental group than those in the control group( P < 0. 05).( 2) PBEF protein expressed in alveolar epithelial cell,bronchial epithelial cells,pulmonary vascular endothelial cell and inflammatory cells predominantly. The IOD on the 3th and 7th day were significantly increased in the experimental group compared with those in the control group( P < 0. 05).( 3) The PBEF mRNA expressions of PBEF on the 3th and 7th day were significantly increased in the experimental group compared with those in the control group and the protein expressions of PBEF were significantly increased in the experimental group compared with those in the control group( P < 0. 05). Conclusion PBEF is likely to play an active role during the developing of hyperoxia-induced BPD.
出处
《中国临床新医学》
2016年第6期497-502,共6页
CHINESE JOURNAL OF NEW CLINICAL MEDICINE
