Shinichiro Nishimatsu    Department   Kawasaki Medical School  Kawasaki Medical School, Department of Natural Sciences,    Position   Professor
Article types	原著
Language	English
Peer review	Peer reviewed
Title	Paraquat-induced gene expression in rat lung tissues using a differential display reverse transcription-polymerase chain reaction.
Journal	Formal name：Archives of toxicology Abbreviation：Arch Toxicol ISSN code：03405761/14320738
Volume, Issue, Page	76(9),pp.530-537
Author and coauthor	Tomita Masafumi, Nohno Tsutomu, Okuyama Toshiko, Nishimatsu Shin-ichiro, Adachi Junko
Publication date	2002/09
Summary	Increased formation of reactive oxygen species is a cause of paraquat (PQ)-induced injury and also provides a link between the signaling pathways and transcriptional events that regulate the expression of a large number of genes. However, the molecular mechanisms involved in PQ-induced injury remain unclear. To investigate the changes in gene expression at the onset of PQ injury, we used the differential display-polymerase chain reaction (PCR) method. Rats were treated intra-peritoneally with 20 mg/kg PQ, and after 3 h the lungs were immediately excised. Samples of mRNA from normal and treated rats were used to prepare radiolabeled cDNAs, which were electrophoresed. Then the transcription levels were compared. We isolated 26 fragments of cDNA that were potentially affected by PQ, and determined their nucleotide sequences. Six clones of interest were selected and analyzed further. The reverse transcript-PCR based on their sequence information confirmed the differential expression for five clones: four clones were up-regulated and one was down-regulated. We were particularly interested in two genes that had homology with the known gene: TATA box-binding protein-associated factor, RNA polymerase II, B, 150 kDa (TAFIIB), and a candidate gene for lipodystrophy, Lpin2. Both genes were significantly up-regulated within 3 h of PQ intake and the stimulation continued during our 24-h observation period. In addition, up-regulation of Lpin2 was observed in the lungs, but not in the liver and kidneys. In situ hybridization using lung sections showed that the expression of both genes was strongly visualized in Clara cells and in alveolar macrophages. These findings suggest a stimulation of transcription levels and changes in lipid metabolism in Clara cells and in macrophages in the lungs, which result in their playing a crucial role at the onset of PQ-driven pulmonary injury.
DOI	10.1007/s00204-002-0379-x
Document No.	12242611