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

本研究旨在阐明细胞在分子水平上的应激反应，分析围手术期药物对应激反应的修饰，并鉴定围手术期药物诱导的基因表达变化。所得结果总结如下1。CRF-1受体拮抗剂可抑制剖腹手术所致大鼠血浆去甲肾上腺素浓度升高。提示中枢神经系统中可能存在抑制手术损伤引起的交感神经反应的通路。通过稳定转染与荧光素酶基因相关的POMC基因启动子的垂体肿瘤细胞株（AtT20PL），分析围手术期使用药物对POMC基因表达的影响。结果表明，钙通道阻滞剂和局部麻醉剂可增强crh诱导的POMC基因表达。异氟醚是挥发性麻醉药的原型，它抑制大脑皮层血清素的释放，这可能与异氟醚的镇静作用有关。另一方面，氧化亚氮可增加脊髓中血清素的释放，这可能与氧化亚氮在脊髓水平的抗痛觉作用有关。此外，对痛觉肽受体敲除小鼠的分析表明，痛觉肽受体不参与挥发性麻醉剂的麻醉作用，但参与氧化亚氮的抗痛觉肽作用。不同药物对缺氧诱导因子（HIF- 1）介导的缺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激活。鉴定吗啡诱导的t淋巴细胞基因表达变化。结果表明，吗啡可增加细胞凋亡相关基因的表达。

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.