BPD新生大鼠TRF1、TRF2表达及其作用  

The expression and effect of TRF1 and TRF2 in neonatal rats with bronchopulmonary dysplasia

作　　者：高思洋 富建华[1] 薛辛东[1] Gao Siyang;Fu Jianhua;Xue Xindong(Neonatal Department, Shengjing Hospital of China Medical University, Shenyang 110004, China)

机构地区：[1]中国医科大学附属盛京医院新生儿科,沈阳110004

出　　处：《国际儿科学杂志》2019年第5期375-379,共5页International Journal of Pediatrics

基　　金：国家自然科学基金资助项目(81571479、81471489);辽宁省教育厅重点实验室基础研究项目(LZ2015070).

摘　　要：目的研究端粒重复序列结合因子1(telomeric repeat binding factor 1,TRF1)和TRF2在新生大鼠支气管肺发育不良(bronchopulmonary dysplasia,BPD)发生、发展过程中的动态表达规律,阐明其在BPD肺泡发育不良中的作用。方法采用吸入氧浓度为85%的新生SD大鼠制备新生大鼠BPD模型(n=40),对照组采用吸入空气的新生SD大鼠(n=40),每组分别于实验后1 d、3 d、7 d、14 d选取10只处死,收集肺组织标本,进行肺组织HE染色观察病理学改变,采用辐射状肺泡计数(RAC)评价肺泡发育程度,分别应用免疫组织化学法观察TRF1、TRF2定位及表达,蛋白免疫印迹法及实时荧光定量PCR(RT-PCR)检测肺组织TRF1和TRF2蛋白及基因的表达水平。结果免疫组织化学染色可见,BPD组TRF1主要定位于肺泡上皮细胞和支气管上皮细胞的胞核内,TRF2蛋白在肺泡上皮细胞和支气管上皮细胞的胞核、胞浆均有明显定位,且较对照组表达强度增强。与对照组相比,BPD组TRF1、TRF2蛋白表达从1d开始增高(TRF1:对照组0.163±0.022,BPD组0.251±0.022,P<0.05;TRF2:对照组0.156±0.012,BPD组0.240±0.018,P<0.05),趋势持续到14 d(TRF1:对照组0.193±0.024,BPD组0.230±0.025,P<0.05;TRF2:对照组0.225±0.017,BPD组0.350±0.012,P<0.05)。TRF1、TRF2的mRNA表达水平在高氧暴露1~7 d持续增加(TRF1:对照组0.946±0.028,BPD组1.590±0.228,P<0.05;TRF2:对照组0.834±0.083,BPD组1.783±0.262,P<0.05),但在14d时下降(TRF1:对照组2.217±0.225,BPD组1.259±0.217,P<0.05;TRF2:对照组2.143±0.250,BPD组0.997±0.199,P<0.05)。结论在BPD发生发展阶段,TRF1、TRF2作为端粒长度的负性调控因子,蛋白水平均呈表达上调状态,提示其可能参与了BPD肺泡发育不良的病理过程。Objective To investigate the dynamic expression of telomere repeat binding factor 1(TRF1)and telomeric repeat binding factor 2(TRF2)in the development and progression of bronchopulmonary dysplasia(BPD)in neonatal rats and to clarify its role in BPD alveolar dysplasia. Methods The neonatal rat BPD model(n=40)was induced by using neonatal SD rats with inhaled oxygen concentration of 85%.The control group was prepared by inhaled air(n=40). In the two groups, 10 rats were randomly selected from 1 day, 3 days, 7 days, and 14 days after the experiment.The lung tissue samples were collected, HE staining was performed to observe the pathological changes, and the alveolar development degree was evaluated by radial alveolar counting(RAC). Immunohistochemistry was used to observe the localization and expression of TRF1 and TRF2.Western Blot and real-time quantitative PCR(RT-PCR)were used to detect the expression levels of TRF1 and TRF2 proteins and genes in lung tissue. Results Immunohistochemical staining showed that TRF1 was mainly localized in the nucleus of alveolar epithelial cells and bronchial epithelial cells.TRF2 protein was found in the nucleus and cytoplasm of alveolar epithelial cells and bronchial epithelial cells.The expression was significantly higher than that of the control group.Compared with the control group, the TRF1 and TRF2 proteins increased significantly from 1d(TRF1 in control group: 0.163±0.022, in model group: 0.251±0.022;TRF2 in control group: 0.156±0.012, in model group: 0.240±0.018)to 14d(TRF1 in control group: 0.193±0.024, in model group: 0.230±0.025;TRF2 in control group: 0.225±0.017, in model group: 0.350±0.012)rather than the control group(P<0.05). The mRNA expression levels of TRF1 and TRF2 increased continuously from 1d to 7d of hyperoxia exposure(TRF1 in control group: 0.946±0.028, in model group: 1.590±0.228;TRF2 in control group: 0.834±0.083, in model group: 1.783±0.262) and decreased at 14d(TRF1 in control group: 2.217±0.225, in model group: 1.259±0.217, P<0.05;TRF2 in

关 键 词：端粒重复序列结合因子 端粒损伤 支气管肺发育不良 新生大鼠 

分 类 号：R722.1[医药卫生—儿科] R-332[医药卫生—临床医学]