权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Identification and their function of paraquat-induced unknown genes in rat lungs.

百草枯诱导的大鼠肺部未知基因的鉴定及其功能。

基本信息

批准号：
13670439
负责人：
TOMITA Masafumi
金额：
$ 1.15万
依托单位：
Kawasaki Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670439/
关键词：
Paraquat Pulmonary injury Oxidative stress Differential display gene expression Free radical cDNA array Real-time PCR differential display superoxide free radical Clara cell TAFIIB Lpin2 ラット

项目摘要

The aim of the present work was to evaluate the implications for genes that are regulated by oxidative stress at the early stage of PQ exposure in rat lungs. Rats were treated with 20mg/kg PQ for 3 h, and the lungs were immediately excised. Significant pulmonary injury was observed after 2 weeks under our experimental conditions. 1)In the first year, we were particularly interested in two genes, TAFIIB and Lpin2. They were strongly visualized in Clara cells and in alveolar macrophages. These findings suggest the possibility that transcription levels and lipid metabolism are activated in Clara cells and in macrophages, resulting in their playing a crucial role at the onset of PQ-driven pulmonary injury. 2)In the second year, 19 differentiated clones were isolated by differential display-PCR method. Four clones were finally determined to be significantly affected. They were plasma phospholipid transfer protein(PLTP), lactrophiline, all-spectrin and one unknown gene. We showed the distribution of mRNA expression of lactrophiline in lung tissues. 3)In the third year, using cDNA array membrane, we found that PQ exposure gave 29 genes differentially expressed(>1.5-fold). Some positive genes were further validated and quantitated with real-time RT-PCR. The expressions of PDGF, thioredoxin, glutathione S-transferase, SOD were significantly up-regulated, suggesting that the oxygen free radicals were major contributors to the onset of PQ poisoning. In addition, some CYP members (CYP2C6,2C7,2C12,2C13) were remarkably affected. The stimulation of expressions was peculiar to lungs, and not to liver and kidneys. These results suggest that they are involved in the onset of PQ-induced serious pulmonary injury.

本研究的目的是评估PQ暴露对大鼠肺部早期氧化应激调控基因的影响。大鼠以20mg/kg PQ处理3 h，立即切除肺。在我们的实验条件下，2周后观察到明显的肺损伤。1)在第一年，我们对TAFIIB和Lpin2两个基因特别感兴趣。它们在Clara细胞和肺泡巨噬细胞中可见。这些发现表明，Clara细胞和巨噬细胞的转录水平和脂质代谢可能被激活，导致它们在pq驱动的肺损伤发病中起关键作用。2)第二年，用差异显示pcr法分离到19个分化无性系。最终确定有4个无性系受到显著影响。它们是血浆磷脂转移蛋白（PLTP）、嗜乳氨酸、全谱蛋白和一个未知基因。我们显示了嗜乳碱mRNA在肺组织中的表达分布。3)第三年，利用cDNA阵列膜，我们发现PQ暴露导致29个基因差异表达（>1.5倍）。部分阳性基因通过实时RT-PCR进一步验证和定量。PDGF、硫氧还蛋白（thioredoxin）、谷胱甘肽s -转移酶（glutathione S-transferase）、SOD表达显著上调，提示氧自由基是PQ中毒发生的主要原因。此外，一些CYP成员（CYP2C6、2C7、2C12、2C13）明显受到影响。表达的刺激是肺所特有的，而不是肝和肾。这些结果表明它们参与了pq诱导的严重肺损伤的发生。