【摘要】 目的:探讨SPOCK2基因在高氧肺损伤新生儿支气管肺发育不良(bronchopulmonary dysplasia,BPD)大鼠模型中的表达意义。方法:利用高氧肺损伤诱导BPD动物模型,将新生SD大鼠随机分成空气对照组、高氧模型组。HE染色观察肺组织病理改变,q PCR法检测肺组织中SPOCK2 m RNA的表达水平,免疫组化法测定肺SPOCK2蛋白的表达。同时在体外利用高氧刺激A549人肺上皮细胞,q PCR法检测细胞SPOCK2 m RNA的表达水平。结果:成功构建BPD新生大鼠模型。HE染色显示高氧模型组肺组织均产生类似BPD的病理损伤,并且生后10、14 d高氧模型组大鼠肺组织辐射状肺泡计数(pulmonary radical alveolar counts,RAC)值较空气对照组显著减少,差异有统计学意义(P<0.05)。免疫组化显示空气对照组大鼠肺组织中SPOCK2蛋白表达量高于高氧模型组,生后10、14 d空气对照组肺组织SPOCK2蛋白表达呈阳性。q PCR结果示空气对照组大鼠肺组织中SPOCK2 m RNA表达量随时间递增,于生后18 d时表达量最高,生后24 d时仍处于高值(P<0.05),成年时下降;与空气对照组比较,高氧模型组大鼠肺组织中SPOCK2 m RNA表达量降低,生后10、14 d时两组差异有统计学意义(P<0.05)。高氧模型组A549细胞SPOCK2 m RNA表达量高于空气对照组(P<0.05),且有先上升后下降的趋势。结论:SPOCK2基因可能参与肺发育过程,并可能在高氧刺激时对肺组织起保护作用。更多还原

【Abstract】 Objective:To investigate the expression of SPOCK2 gene in hyperoxia-induced bronchopulmonary dysplasia(BPD)models of neonatal rats. Methods:BPD models were induced by 85% O2 exposure. Newborn SD rats were randomly divided into the air group and the hyperoxia(85% oxygen)group. The pathological changes of pulmonary tissues were observed by hematoxylin-eosin(HE)staining. The expressions of SPOCK2 m RNA were detected by real-time quantitative PCR. The expression of SPOCK2 protein was detected by immunohistochemistry. Meanwhile,A549 lung epithelial cells were stimulated by hyperoxia in vitro and the expression levels of SPOCK2 m RNA were detected by q PCR. Results:BPD neonatal rat model was successfully constructed. The pathological changes of BPD were found in the pulmonary tissues of the model group by HE staining. The value of pulmonary radical alveolar counts(RAC)in the model group was significantly lower than that of the control group at 10 d and 14 d after birth(P < 0.05). As shown with immunohistochemistry,the expression of SPOCK2 protein in the lung tissues of rats in the control group was higher than that in the model group,especially at 10 d and 14 d after birth. The expression of SPOCK2 m RNA in the lung tissues of rats in the control group was higher than that in the model group,which reached its peak on 18 d after birth and decreased in adulthood. Compared with the control group,the expression of SPOCK2 m RNA in the model group decreased. There were significant difterences befween two groups at 10 d and 14 d after birth(P < 0.05). The expression of A549 m RNA in the model A549 cells was higher than that in the control(P <0.05). Conclusion:SPOCK2 gene may be involved in pulmonary development process,and may protect lung tissue during hyperoxia stimulation.更多还原