转化生长因子-β1、Clara细胞分泌蛋白16、Ⅱ型肺泡细胞表面抗原6与支气管肺发育不良的相关性研究

Relationships between the expressions of transforming growth factor-β1,Clara cell secretory protein 16,and type Ⅱ alveolar cell surface antigen 6 in lung cell apoptosis in rats with bronchopulmonary dysplasia

目的探讨转化生长因子-β1(transforming growth factor-β1,TGF-β1)、克拉拉细胞分泌蛋白16(clara cell secretory protein,CC16)、Ⅱ型肺泡细胞表面抗原6 (krebs yon denlungen-6,KL-6)在支气管肺发育不良(bronchopulmonary dysplasia,BPD)大鼠肺组织中的表达,以及这些蛋白的表达与肺细胞凋亡的关系。方法按随机数字表法随机将SD新生鼠随机分为对照组、模型组,模型组持续暴露在80%~85%氧中构建BPD模型,对照组始终暴露在空气中。每组在7 d,14 d,21 d时(简称为7 d模型组、14 d模型组、21 d模型组)随机选出8只新生鼠处死,留取5 m L血清。HE染色观察各组肺组织病理学改变,并分析辐射状肺泡计数(radial alveolar count,ROC),肺泡平均截距(mean linear intercept,MLI)。TUNEL染色检测各组肺组织中的细胞凋亡。ELISA检测各组新生鼠的血清中TGF-β1、CC16、KL-6的水平。Western blot检测各组肺组织中TGF-β1、CC16、KL-6的表达水平。同时分析肺组织中TGF-β1、CC16、KL-6的表达与肺组织细胞凋亡率的关系。结果与正常组相比,模型组肺组织随暴露氧时间的延长BPD损伤加剧,RAC、血清中CC16的水平以及肺组织中CC16的表达明显减少,MLI、细胞凋亡率、血清中TGF-β1、KL-6的水平,肺组织中TGF-β1、KL-6的表达明显增加,差异均具有统计学意义(P<0.05)。相关性分析显示,模型组新生鼠肺组织中TGF-β1、KL-6的表达与肺组织的细胞凋亡率呈正相关(r=0.977、0.970,均P=0.000),CC16的表达与细胞凋亡率呈现负相关(r=-0.982,P=0.000)。结论肺组织中TGF-β1、CC16、KL-6的表达与BPD肺组织的细胞凋亡密切相关。

Objective To investigate the expressions of transforming growth factor-β1(TGF-β1),Clara cell secretory protein 16(CC16),and typeⅡalveolar cell surface antigen 6(Krebs Yon Denlungen-6;KL-6)in the lung tissue of bronchopulmonary dysplasia(BPD)-model rats and the relationship between the expressions of these proteins and lung cell apoptosis.Methods Newborn Sprague-Dawley rats were randomly divided into the control and model groups according to the random number table method.Rats in the model group were continuously exposed to 80%~85%oxygen to construct the BPD model,and rats in the control group were always exposed to regular air.At 7,14,and 21 days(i.e.,the 7 d model,14 d model,and 21 d model groups),eight newborn rats were randomly selected and euthanized,and 5 m L of serum was retained.Hematoxylin and eosin staining was used to observe the lung histopathological changes in each group,and the radial alveolar count(RAC)and mean alveolar intercept were analyzed.TUNEL staining was used to detect apoptosis in the lung tissues of each group.Enzyme-linked immunosorbent assays were used to detect the TGF-β1,CC16 and KL-6 serum levels of newborn rats in each group.Western blot was used to detect the TGF-β1,CC16 and KL-6 expression levels in the lung tissue of each group.The relationships between the TGF-β1,CC16 and KL-6 expressions and the apoptosis rates in the lung tissue were analyzed.Results Compared with that of the normal group,the BPD injury to the lung tissue in the model group increased as the 80%~85%oxygen exposure was prolonged.The serum levels of RAC and CC16 and the CC16 expression in the lung tissue were significantly reduced.At the serum level,the TGF-β1 and KL-6 expressions in the lung tissue were increased significantly(P<0.05).Correlation analysis showed that the TGF-β1 and KL-6 expressions in the lung tissue of the neonatal rats in the model group were positively correlated with the apoptosis rate in the lung tissue(r=0.977,0.970,P=0.000),and CC16 expression was correlated with apoptosis.The mortality rate showed a negative correlation(r=-0.982,P=0.000).Conclusions TGF-β1,CC16,and KL-6 expression in lung tissue was closely correlated with apoptosis in the lung tissues of BPD-model rats.