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目的探讨增殖细胞核抗原(proliferatingeellnuelearantigen,PCNA)在CLD新生鼠肺成纤维细胞中的动态表达及意义。方法足月新生SD大鼠于生后12h内分别持续吸入85%高氧和空气,于3、7、14、21天随机处死后动物,无菌条件下采集肺组织,进行LF细胞的原代培养,应用MTT法检测细胞增殖活力。免疫组化法检测PCNA蛋白表达;Real-time PCR法检测PCNAmRNA含量。结果 MTT结果显示,高氧使得LF增殖活力增强;生后3d空气组和高氧组PCNA蛋白及mRNA的表达无统计学差异。生后7d时高氧组PCNA蛋白及mRNA的表达较空气组增加(P<0.05);14d、21d时增加明显(P<0.01)。结论体内高氧使LF增殖活力增加,最终导致肺间质纤维化。
Objective To investigate the dynamic expression and significance of proliferating cell nuclear antigen (PCNA) in lung fibroblasts of neonatal rats with CLD. Methods SD rats of term newborn were inhaled 85% of hyperoxia and air respectively within 12 hours after birth. Animals were randomly sacrificed on days 3, 7, 14, and 21. After aseptic conditions, lung tissues were collected for primary generation of LF cells Culture, application of MTT assay cell proliferation activity. The expression of PCNA protein was detected by immunohistochemistry. The content of PCNA mRNA was detected by Real-time PCR. Results The results of MTT showed that the hyperoxia increased the proliferative activity of LF. There was no significant difference in the expression of PCNA protein and mRNA between the air group and the hyperoxia group after birth. On the 7th day after birth, the expression of PCNA protein and mRNA in hyperoxia group was higher than that in air group (P <0.05), and increased significantly on the 14th and 21st day (P <0.01). Conclusion High oxygen in vivo increases LF proliferation activity and eventually leads to pulmonary interstitial fibrosis.