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高氧诱导新生大鼠急性肺损伤中 Shh 和 Ptc1 的上调。

Upregulation of Shh and Ptc1 in hyperoxia‑induced acute lung injury in neonatal rats.

机构信息

PICU, Children's Hospital of Chongqing Medical University, Yu Zhong, Chongqing 400014, PR China.

出版信息

Mol Med Rep. 2012 Aug;6(2):297-302. doi: 10.3892/mmr.2012.929. Epub 2012 May 28.

DOI:10.3892/mmr.2012.929
PMID:22641469
Abstract

The aim of the present study was to observe the expression of sonic hedgehog (Shh) and Ptc signaling molecules in the lungs of newborn rats exposed to prolonged hyperoxia, and to explore the role of the SHH signaling pathway in hyperoxia‑induced lung injury. Newborn Sprague-Dawley rat pups were placed in chambers containing room air or oxygen above 95% for 14 days following birth. The rats were sacrificed after 3, 7 or 14 days and their lungs were removed. Sections were fixed and subjected to hematoxylin and eosin (H&E) staining. Shh and Ptc1 were quantitated by immunohistochemistry. The total RNA and protein were also extracted from lung tissue; real-time PCR (RT-PCR) and western blot analysis were utilized to assess the mRNA and protein expression of Shh and Ptc1. H&E staining demonstrated significant histomorphological changes in the hyperoxia‑exposed lungs at 3, 7 and 14 days of age. The results of the immunohistochemistry, RT-PCR and western blot analysis demonstrated that the expression of Shh was significantly higher in the hyperoxia-exposed lungs at 3, 7 and 14 days, while Ptc1 was significantly elevated at 7 and 14 days. Exposure of the neonatal rat lung to prolonged hyperoxia resulted in acute lung injury and histomorphological changes. Shh and Ptc1 were upregulated in a time-dependent manner in the course of hyperoxia-induced lung injury. The SHH signal pathway may be involved in the pathogenesis of hyperoxia-induced lung injury. This is the first evidence that in vivo hyperoxia induces activation of the SHH signal transduction pathway in newborn lung.

摘要

本研究旨在观察 sonic hedgehog(Shh)和 Ptc 信号分子在长时间高氧暴露新生大鼠肺中的表达,探讨 SHH 信号通路在高氧诱导肺损伤中的作用。新生 Sprague-Dawley 大鼠在出生后第 14 天被置于含有室内空气或高于 95%氧气的室中。在 3、7 或 14 天后处死大鼠并取出其肺。对肺组织进行苏木精和伊红(H&E)染色。通过免疫组织化学法对 Shh 和 Ptc1 进行定量。还从肺组织中提取总 RNA 和蛋白质;利用实时 PCR(RT-PCR)和 Western blot 分析评估 Shh 和 Ptc1 的 mRNA 和蛋白表达。H&E 染色显示,在高氧暴露 3、7 和 14 天的大鼠肺中,组织形态学发生了明显变化。免疫组织化学、RT-PCR 和 Western blot 分析结果表明,在高氧暴露 3、7 和 14 天的大鼠肺中,Shh 的表达显著升高,而 Ptc1 在 7 和 14 天显著升高。新生大鼠肺长时间暴露于高氧会导致急性肺损伤和组织形态学改变。在高氧诱导的肺损伤过程中,Shh 和 Ptc1 呈时间依赖性上调。SHH 信号通路可能参与高氧诱导的肺损伤的发病机制。这是体内高氧诱导新生肺中 SHH 信号转导通路激活的第一个证据。

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