HIDAKA Kazuo Kawasaki Medical School, Biochemistry, Lecturer, 医学部, 講師 (00069064)
NISHIMATSU Shin-ichiro Kawasaki Medical School, Molecular Biology, Lecturer, 医学部, 講師 (20222185)
NOHNO Tsutomu Kawasaki Medical School, Molecular Biology, Associate Professor, 医学部, 助教授 (20098619)
|Budget Amount *help
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2003: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2002: ¥1,000,000 (Direct Cost: ¥1,000,000)
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.