【作者】 章功良；

【导师】 赵乐章； 张景行；

【作者基本信息】 安徽医科大学 ， 生理学， 2004， 硕士

【摘要】 本研究采用脑立体定位、核团插管、微量注射、核团毁损、多导睡眠描记、免疫组织化学染色和行为活动测定等方法，研究P物质(substance P，SP)和5-羟色胺(serotonin，5-hydroxytryptamine，5-HT)对大鼠睡眠-觉醒周期的影响及其机制以及大鼠vLPO化学毁损后行为活动改变。 1 侧脑室注射P物质对大鼠睡眠—觉醒周期的影响 1．1大鼠侧脑室内微量注射SP(100nmol)后睡眠时间增加，觉醒时间减少；而在侧脑室内微量注射SP受体阻断剂(N-acetyl-L-tryptophan 3，5-bis(trifluoromethyl)-benzyl ester，100nmol)后睡眠时间减少，觉醒时间增加。 1．2大鼠腹外侧视前区(ventrolateral preoptic area，vLPO)内微量注射兴奋性神经细胞毒剂海人酸(kainic acid，KA，0.4μg)后，睡眠时间减少，觉醒时间增加；KA毁损vLPO后，侧脑室注射SP，SP原有的促眠作用未见出现。 1．3侧脑室注射SP(100nmol)，应用免疫组化检测，观察到vLPO Fos蛋白表达较正常对照组增加。 2 腹外侧视前区SP对大鼠睡眠-觉醒周期的影响 2．1大鼠vLPO内微量注射SP(10nmol)后与对照组比较睡眠时间增加，特别是深慢波睡眠时间明显增加，觉醒时间减少；而在vLPO内微量注射SP受体(NK-1)阻断剂(N-acetyl-L-tryptophan 3，5-bis(trifluoromethyl)-benzyl ester，10nmol)后睡眠时间减少，觉醒时间增加。 2．2大鼠vLPO内微量注射磷脂酶C(PLC)抑制剂U73122(20nmol)后睡眠时间减少，觉醒时间增加；大鼠vLPO内微量注射U73122(20nmol)阻断SP受体信号跨膜转导，30分钟后再在vLPO微量注射SP(10nmol)，SP原有的促眠作用未见出安徽医科大学硕士学位论文现。2.3大鼠vLPO内微量注射GABA合成关键酶抑制剂3一琉基丙酸(3一MP，5瑰)，注射当日睡眠时间轻度减少，第二日睡眠时间明显减少，觉醒时间明显增加，第三、四日睡眠和觉醒时间逐渐恢复至正常;在大鼠vLPO内微量注射3一MP后第二日，vLPO内微量注射SP(10nm。1)，SP原有的促眠作用未见出现。2.4完全剥夺大鼠24小时睡眠后，应用免疫组化检测，vLPO SP染色较正常对照组增加。3腹外侧视前区5一HT对大鼠睡眠一觉醒周期的影响3.1大鼠vLPO内微量注射小剂量5一HT前体5一轻色胺酸(5一HTP，0.5陀)对大鼠睡眠一觉醒周期无明显影响;vLPO内微量注射5一HTP(1魄)或5一HT转运体抑制剂氟西汀(6协g)后睡眠时间减少，觉醒时间增加;而在vLPO内微量注射非特异性5一HT受体阻断剂二甲基麦角新碱(MS，0.5拼g)后睡眠时间增加，觉醒时间减少。3.25一经色胺酸的促醒作用和MS的促眠作用有时间相关性。3，3大鼠vLPO内微量注射KA毁损vLPO神经元后，通过开野实验和高架十字迷宫实验观察大鼠的行为活动，发现大鼠的探究行为等活动有增加趋势，但与对照组相比没有统计学上显著性差异。本研究表明:1.P物质是一种中枢性促眠物质，但必须通过vLPO介导。2.P物质在腹外侧视前区具有促进大鼠睡眠的作用，特别是深慢波睡眠;SP的促 眠作用通过GABA能神经元介导。3.完全性睡眠剥夺后SP表达上调。4.5一经色胺在腹外侧视前区具有促进大鼠觉醒的作用。5.腹外侧视前区具有抑制动物行为活动的效应，但不具有统计学上的显著性差 异。更多还原

【Abstract】 The present study investigated the effects and mechanism of substance P (SP) and serotonin (5-HT) on sleep-wakefulness cycle of rats and the role of in the ventrolateral preoptic area (vLPO) on behavioral activities by using brain stereotaxic, nucleus spile, microinjection, lesion of nucleus, polysomnography (PSG), immunohistochemistry and behavioral test.1 The effects of injecting SP into lateral cerebral ventricle on percent of sleep-wakefulness in rats1.1 Microinjection of SP (100nmol) into lateral cerebral ventricle enhanced sleep and reduced wakefulness compared with control, while SP receptor antagonist(N-acetyl-L-tryptophan 3, 5~bis(trifluoromethyl)-benzyl ester, 100nmol)had reverse effect.1.2 Microinjection of kainic acid (a kind of neural excitotoxic agent, KA, 0.4 g) into vLPO reduced sleep and enhanced wakefulness compared with control, and SP central sleep-promoting effect disappeared when vLPO lesion by KA.1.3 The expression of Fos in the vLPO increased compared with control when microinjection of SP (100nmol) into lateral cerebral ventricle.2 The effects of SP in the vLPO on percent of sleep-wakefulness in rats2.1 Microinjection of SP (10nmol) into vLPO enhanced sleep, especially deep slow wave sleep, and reduced wakefulness compared with control, while SP receptor (NK-1) antagonist (N-acetyl-L-tryptophan3, 5-bis (trifluoromethyl) -benzyl ester, lOnmol)had reverse effect.2.2 Microinjection of U73122, a kind of phospholipase C (PLC) inhibitor, reduced sleep and inhanced wakefulness; and SP sleep-promoting effect didn’t show when pre-treating with U73122 thirty minutes before.2.3 Microinjection of 3-mercaptopropionic acid (3-MP, a kind of glutamate decarboxylase inhibitor) into vLPO. On the day of microinjection, sleep only decreased a little. On the second day, sleep marked decreased and wakefulness marked increased. On the third and fourth day, sleep and wakefulness stages resumed to normal. SP sleep-promoting effect didn’t show when pre-treating with 3-MP on the previous day.2.4 The expression of SP increased in vLPO in total sleep deprivation of 24 hours compared with control with immunocytochemistry.3 Effects of serotonin in the vLPO on percent of sleep-wakefulness cycle in rats3.1 There was no significant effect when microinjection of 5-hydroxytryptophan (5-HTP, 0.5 |ig, serotonin precursor) into vLPO on sleep-wakefulness cycle, but microinjection of 5-HTP (1 g) and fluoxetine (6 g) led to wakefuness increased and sleep decreased. Microinjection of non-selective serotonin receptor antagonist methysergide (MS) led to the opposited effect.3.2 The percent of sleep-wakfulness caused by 5-HTP or MS were significant assiociated with time.3.3 Compared with control, the behavior activities increased when vLPO lesion by KA using open-field test and elevated plus maze test, but there were no statistaical difference between lesion group and control.The present study indicated:1. Substance P is a central sleep-promoting substance and vLPO is a target of central SP acting on.2. Substance P has sleep-promoting effect in the vLPO of rats, especially deep slow wave sleep. The sleep-promoting effect of SP is mediated by GABAergic neurons in vLPO.3. Total sleep deprivation up-regulates the expression of SP.4. Serotonin has wakefulness-promoting effect in the vLPO of rats.5. Ventrolateral preoptic area could inhibit rat behavioral activities, but there were no statistaical difference.更多还原