Hara S, Kobayashi M, Kuriiwa F, Mukai T, Mizukami H
Department of Forensic Medicine, Tokyo Medical University , Tokyo , Japan.
Free Radic Res. 2014 Nov;48(11):1322-33. doi: 10.3109/10715762.2014.951842. Epub 2014 Sep 2.
Previous studies have suggested that carbon monoxide (CO) poisoning stimulates cAMP production via purine P2Y11-like receptors in the rat striatum, activating cAMP signaling pathways, resulting in hydroxyl radical ((•)OH) production. Extracellular ATP was thought likely to trigger the cascade, but the present study has failed to demonstrate a clear increase in the extracellular ATP due to CO poisoning. The CO-induced (•)OH production was attenuated by the P2Y11 receptor antagonist NF157, in parallel with its abilities to suppress the CO-induced cAMP production. The (•)OH production was more strongly suppressed by a non-selective P2 receptor antagonist, PPADS, which had no effect on cAMP production. More selective antagonists toward the respective P2 receptors susceptible to PPADS, including NF279, had little or no effect on the CO-induced (•)OH production. The intrastriatal administration of exogenous ATP dose-dependently stimulated (•)OH production, which was dose-dependently antagonized by PPADS and NF279 but not by NF157. Exogenous GTP and CTP dose-dependently stimulated (•)OH production, though less potently. The GTP-induced (•)OH production was susceptible to both of NF279 and PPADS, but the CTP-induced (•)OH production was resistant to PPADS. The mechanism of (•)OH production may differ between CO poisoning and exogenous ATP, while multiple P2 receptors could participate in (•)OH production. The CO-induced (•)OH production was susceptible to the inhibition of NADPH oxidase, but not xanthine oxidase. Also, the NADPH oxidase inhibition suppressed (•)OH production induced by forskolin, a stimulator of intracellular cAMP formation. It is likely that (•)OH is produced by NADPH oxidase activation via cAMP signaling pathways during CO poisoning.
先前的研究表明,一氧化碳(CO)中毒通过大鼠纹状体中的嘌呤P2Y11样受体刺激环磷酸腺苷(cAMP)的产生,激活cAMP信号通路,导致羟自由基(•OH)的产生。细胞外ATP被认为可能触发这一系列反应,但本研究未能证明CO中毒导致细胞外ATP明显增加。P2Y11受体拮抗剂NF157可减弱CO诱导的•OH产生,同时其抑制CO诱导的cAMP产生的能力也相应减弱。非选择性P2受体拮抗剂PPADS对•OH产生的抑制作用更强,而对cAMP产生没有影响。对PPADS敏感的各P2受体的更具选择性的拮抗剂,包括NF279,对CO诱导的•OH产生几乎没有影响。纹状体内注射外源性ATP可剂量依赖性地刺激•OH产生,PPADS和NF279可剂量依赖性地拮抗该作用,但NF157无此作用。外源性鸟苷三磷酸(GTP)和胞苷三磷酸(CTP)可剂量依赖性地刺激•OH产生,但其效力较弱。GTP诱导的•OH产生对NF279和PPADS均敏感,但CTP诱导的•OH产生对PPADS有抗性。CO中毒和外源性ATP诱导•OH产生的机制可能不同,而多种P2受体可能参与•OH的产生。CO诱导的•OH产生对烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶的抑制敏感,但对黄嘌呤氧化酶不敏感。此外,NADPH氧化酶抑制可抑制由细胞内cAMP形成刺激剂福司可林诱导的•OH产生。在CO中毒期间,•OH可能是通过cAMP信号通路激活NADPH氧化酶产生的。