论文部分内容阅读
目的:研究长链非编码RNA牛磺酸上调基因1(taurine upregulation gene 1，Tug1)在新生大鼠支气管肺发育不良(bronchopulmonary dysplasia，BPD)模型中的表达，并从肺组织形态学及肺泡发育成熟度两方面探究长链非编码RNA Tug1与肺发育的相关性，为阐明BPD中肺发育障碍的病理机制奠定基础。方法:采用高氧诱导新生SD大鼠制备BPD模型(氧浓度为85%，n n＝40)，对照组为空气环境正常生长的新生SD大鼠(氧浓度21%，n n＝40)。每组分别于生后1、3、7、14 d随机取10只采集肺组织样本。通过苏木精-伊红染色观察肺组织形态学改变，应用辐射状肺泡计数法(RAC)评估肺泡发育成熟度，利用实时荧光定量PCR(RT-PCR)检测肺组织中长链非编码RNA Tug1及血小板源性生长因子受体α(platelet derived growth factor receptor alpha，Pdgfra)的基因水平，用蛋白免疫印迹技术检测Pdgfra的蛋白表达情况。n 结果:在对照组，Tug1及Pdgfra的mRNA表达随日龄增加呈递增趋势，且Tug1的表达与RAC值呈显著正相关(n r＝0.648，n P＝0.007)，与Pdgfra的mRNA表达呈正相关(n r＝0.572，n P＝0.021)，与Pdgfra蛋白表达也呈正相关(n r＝0.755，n P＝0.001)。模型组中，Tug1表达从7 d开始显著低于对照组(0.78±0.20比1.68±0.20，n P＝0.04)，趋势持续至14 d；Pdgfra的mRNA表达从7 d开始显著低于对照组(1.04±0.25比1.62±0.37，n P＝0.002)，趋势持续至14 d。Pdgfra蛋白表达在对照组随日龄增加呈递增趋势，模型组7 d开始显著低于对照组(1.04±0.13比1.62±0.09，n P＝0.04)，趋势持续至14 d。n 结论:长链非编码RNA Tug1与肺发育成熟度密切相关，其表达在正常发育肺组织中呈递增趋势，新生大鼠BPD模型肺组织中Tug1表达下调可能是BPD肺泡发育障碍的发生机制之一。“,”Objective:To study the expression of long non-coding RNA taurine upregulation gene 1 (Tug1) in the model of bronchopulmonary dysplasia (BPD) in newborn rats, and explore the correlation between Tug1 and lung development from the aspects of lung histology and alveolar maturity, so as to lay a foundation for elucidate the pathological mechanism of lung development disorders in BPD.Methods:Newborn SD rats induced by high oxygen were used to prepare BPD model (oxygen concentration was 85%, n n=40). The control group was newborn SD rats growing normally in air environment (oxygen concentration was 21%, n n=40). In each group, ten lung tissue samples were randomly collected at 1, 3, 7 and 14 days after birth.Pulmonary histopathological changes were observed by HE staining, the development of alveolar was evaluated by radial alveolar count(RAC), the expression levels of long non-coding RNA Tug1 and platelet derived growth factor receptor alpha(Pdgfra) in lung tissues were detected by real-time quantitative PCR(RT-PCR), and the protein expression of Pdgfra was detected by western blot.n Results:In the control group, the expression level of Tug1 and Pdgfra mRNA increased with age, and the expression of Tug1 was positively correlated with RAC value(n r=0.701, n P=0.01), as well as mRNA expression of Pdgfra(n r=0.701, n P=0.01). In the model group, Tug1 expression was significantly lower than that in the control group[(1.68±0.20)in the control group, (0.78±0.20)in the model group, n P=0.040] from day 7, and the decreasing trend continued to 14 days.The mRNA expression of Pdgfra in the model was significantly lower than that in the control group[(1.62±0.37)in the control group, (1.04±0.25) in the model group, n P=0.002] from day 7 to day 14.Pdgfra protein expression level in the control group showed an increasing trend with the increase of daily age, and the trend of Pdgfra protein expression in the model group was significantly lower than that in the control group from 7 days to 14 days.n Conclusion:Long non-coding RNA Tug1 expression is increasing in normal lung development, and down regulation of Tug1 expression in neonatal rats exposed to hyperoxia may be one of the mechanisms of alveolar development disorders in BPD.