Analysis and Clinical Application of Gene Expression Changes Induced by Perioperative Stresses

本研究旨在从分子水平阐明细胞应激反应，分析围手术期使用的药物对应激反应的修饰，并鉴定围手术期使用的药物诱导的基因表达变化。结果如下：1. CRF-1受体拮抗剂可抑制大鼠剖腹手术引起的血浆去甲肾上腺素浓度升高。提示中枢神经系统中存在一条抑制手术损伤诱导的交感神经反应的通路。2.采用稳定转染阿黑皮素原（proopiomelanocortin，POMC）基因启动子和荧光素酶基因的垂体瘤细胞株（AtT 20 PL），研究围手术期用药对POMC基因表达的影响。结果表明，钙通道阻滞剂和局麻药可增强CRH诱导的POMC基因表达。3.异氟醚是挥发性麻醉药的原型，可抑制大脑皮层5-羟色胺的释放，这可能与异氟醚的镇静作用有关。另一方面，一氧化二氮显示增加脊髓中5-羟色胺的释放，这被认为参与了一氧化二氮在脊髓水平的抗伤害作用。此外，对孤啡肽受体敲除小鼠的分析表明，孤啡肽受体不参与挥发性麻醉药的麻醉作用，但参与一氧化氮的抗伤害作用。4.不同药物对低氧诱导因子（HIF- 1）介导的低氧诱导基因表达变化的影响。结果表明，血管扩张剂硝普钠和静脉麻醉剂异丙酚可抑制缺氧诱导的HIF-1的活化。5.吗啡诱导T淋巴细胞基因表达的变化。结果表明，吗啡可增加癫痫相关基因的表达。

This research was aimed to clarify cellular stress responses at molecular levels, to analyze modification of stress responses by perioperatively used drugs, and identify gene expression changes induced by perioperatively used drugs. The results obtained are summarized below.1.Increase in plasma norepinephrine concentration induced by laparotomy in rats was suppressed by CRF-1 receptor antagonists. It was suggested that a pathway exists in the central nervous system which inhibits sympathetic nervous responses induced by surgical insults.2.Effects of perioperatively used drugs on the proopiomelanocortin (POMC) gene expression was analyzed by a pituitary tumor cell line (AtT20PL) which was stably transfected with the POMC gene promoter linked with the luciferase gene. It was shown that calcium channel blockers and local anesthetics can potentiate CRH-induced POMC gene expression.3.Isoflurane, a prototype of volatile anesthetics, suppresses serotonin release in the cerebral cortex, which can be related with sedative action of isoflurane. On the other hand, nitrous oxide was shown to increase serotonin release in the spinal cord, which was suggested to be involved in the antinociceptive action of nitrous oxide at the spinal cord level. Furthermore, analysis of nociceptin receptor knockout mice suggested that the nociceptin receptor is not involved in the anesthetic action of volatile anesthetics but contributes to the antinociceptive action of nitrous oxide.4.Effects of various drugs on the hypoxia-induced gene expression changes mediated by hypoxia-inducible factor (HIF- 1). It was shown that a vasodilator sodium nitroprusside and a n intravenous anesthetic agent propofol inhibit hypoxia-induced HIF-1 activation.5.Morphine-induced gene expression changes in T-lymphocytes were identified. It was shown that expression of apoptosis-related genes was increased by morphine.