
高氧暴露对早产鼠肺组织HO-1和GCLC表达的影响
楚晓云, 蔡成, 张潇月, 周慧琳, 孙俊芳, 翁博雯
中国当代儿科杂志 ›› 2019, Vol. 21 ›› Issue (6) : 594-600.
高氧暴露对早产鼠肺组织HO-1和GCLC表达的影响
Effect of hyperoxic exposure on the expression of heme oxygenase-1 and glutamate-L-cysteine ligase catalytic subunit in lung tissue of preterm rats
目的探讨高浓度氧暴露对新生早产大鼠肺组织中血红素加氧酶-1(HO-1)和谷氨酰-L-半胱氨酸连接酶催化亚单位(GCLC)动态表达的影响。方法受孕21 d大鼠行剖宫产取出早产鼠80只,喂养1 d后随机分为空气组和高氧组。空气组早产鼠放置在室内常压空气中饲养,高氧组早产鼠放置在同一室内常压氧箱中(氧浓度85%~95%)饲养,分别于第1、4、7、10、14天,每组取8只大鼠,采集两组早产鼠肺组织标本。采用苏木精-伊红染色法检测两组早产鼠不同时间点肺组织结构的病理变化;采用Western blot技术和RT-qPCR检测两组早产鼠不同时间点肺组织HO-1和GCLC蛋白及mRNA的表达变化。结果与空气组相比,高氧组早产鼠体重下降显著(P < 0.05)。与空气组相比,高氧组早产鼠肺组织病理切片显示肺组织结构紊乱、肺泡间隔增宽、肺泡数目减少和肺泡简单化。高氧组早产鼠肺组织中HO-1 mRNA相对表达量在第7天时低于空气组,在第10、14天时高于空气组(P < 0.05)。高氧组早产鼠肺组织中GCLC mRNA表达量在第1、4、7天时低于空气组,在第10天时高于空气组(P < 0.05)。与空气组比较,高氧组早产鼠肺组织中HO-1蛋白表达水平在各时间点均升高;除第1天外,GCLC蛋白表达水平在其他各时间点均升高(P < 0.05)。结论高氧暴露导致早产鼠生长发育迟缓、肺发育阻滞。早产鼠肺组织HO-1和GCLC蛋白及mRNA的表达变化可能与高氧暴露导致早产鼠肺损伤发病过程相关。
Objective To study the effect of hyperoxic exposure on the dynamic expression of heme oxygenase-1 (HO-1) and glutamate-L-cysteine ligase catalytic subunit (GCLC) in the lung tissue of preterm neonatal rats. Methods Cesarean section was performed for rats on day 21 of gestation to obtain 80 preterm rats, which were randomly divided into air group and hyperoxia group after one day of feeding. The rats in the air group were housed in room air under atmospheric pressure, and those in the hyperoxia group were placed in an atmospheric oxygen tank (oxygen concentration 85%-95%) in the same room. Eight rats each were selected from each group on days 1, 4, 7, 10, and 14, and lung tissue samples were collected. Hematoxylin and eosin staining was used to observe the pathological changes of lung tissue at different time points after air or hyperoxic exposure. Western blot and RT-qPCR were used to measure the protein and mRNA expression of HO-1 and GCLC in the lung tissue of preterm rats at different time points after air or hyperoxic exposure. Results Compared with the air group, the hyperoxia group had a significant reduction in the body weight (P < 0.05). Compared with the air group, the hyperoxia group had structural disorder, widening of alveolar septa, a reduction in the number of alveoli, and simplification of the alveoli on the pathological section of lung tissue. Compared with the air group, the hyperoxia group had significantly lower relative mRNA expression of HO-1 in the lung tissue on day 7 and significantly higher expression on days 10 and 14 (P < 0.05). Compared with the air group, the hyperoxia group had significantly lower mRNA expression of GCLC in the lung tissue on days 1, 4, and 7 and significantly higher expression on day 10 (P < 0.05). Compared with the air group, the hyperoxia group had significantly higher protein expression of HO-1 in the lung tissue on all days, and the protein expression of GCLC had same results as HO-1, except on day 1 (P < 0.05). Conclusions Hyperoxia exposure may lead to growth retardation and lung developmental retardation in preterm rats. Changes in the protein and mRNA expression of HO-1 and GCLC in the lung tissue of preterm rats may be associated with the pathogenesis of hyperoxia-induced lung injury in preterm rats.
高浓度氧 / 血红素加氧酶-1 / 谷氨酰-L-半胱氨酸连接酶催化亚单位 / 早产 / 大鼠
High-concentration oxygen / Heme oxygenase-1 / Glutamate-L-cysteine ligase catalytic subunit / Preterm birth / Rats
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国家自然科学基金(81571467)。