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The effects and mechanism of GABAergic neurons, NOergic neurons, opioid peptide and cyclic adenosine monophosphate in the nucleus reticularis thalami on sleep-wakefulness cycle of rats and the effects and mechanism of the 5-HTergic nerve fibers project from the nucleus raphes dorsalis to RT on sleep-wakefulness cycle of rats were investigated with the methods of brain stereotaxic, nucleus spile, microinjection and polysomngraphy.1. The effects of GABAergic neurons in RT on sleep-wakefulness cycle of rats1.1 Microinjection of 3-mercaptopropionic acid (3-MP, a kind of glutamate decarboxylase inhibitor) into RT. On the day of microinjection, sleep only decreased a litter. On the second day, sleep marked decreased and wakefulness marked increased. On the third and fourth day, sleep and wakefulness stages resumed to normal.1.2 Microinjection of gamma-amino butyric acid (GABA 1.0μg) into RT enhanced sleep and reduced wakefulness compared with control; while microinjection of L-glutamate (L-Glu, 0.2μg) decreased sleep and increased wakefulness; microinjection of bicuculline (BIC, 1.0μg), a GABAA receptor antagonist, enhanced wakefulness and reduced sleep; microinjection of baclofen (BAC, 1.0μg), GABAB receptor agonist, had the same effects as GABA.2. The effects of NOergic neurons in RT on sleep-wakefulness cycle of rats2.1 Microinjection of L-arginine (L-Arg, 0.5μg) into RT decreased sleep compared with control, but there were on statistaical difference between L-Arg group and control; while microinjection of sodium nitroprusside (SNP, 0.2μg), a NO donor into RT, sleep marked decreased and wakefulness marked increased. Microinjection of nitric oxide synthase inhibitor, N-nitro-L-arginine (L-NNA, 2.0μg) into RT enhanced sleep and reduced wakefulness.2.2 After simultaneous microinjection of L-NNA (2.0μg) and SNP (0.2μg) into RT, SNP abolished the sleep-promoting effect of L-NNA compared with L-NNA group; after simultaneous microinjection of L-NNA (2.0μg) and L-Arg(0.5μg) into RT, we found that L-NNA could not blocked the wakefulness-promoting effect of L-Arg.3. The effects of opioid peptide in RT on sleep-wakefulness cycle of rats3.1 Microinjection of morphine sulfate (MOR, 1.0μg) into RT increased wakefulness and decreased sleep compared with control; while microinjection of naloxone hydrochloride (NAL, 1.0μg), the antagonist of opiate receptors, into RT, enhanced sleep and reduced wakefulness.3.2 After simultaneous microinjection of MOR (1.0μg) and NAL (1.0μg) into RT, the wakefulness-promoting effect of MOR and the sleep-promoting effect of NAL were not observed compared with control.4. The effects of cAMP in RT on sleep-wakefulness cycle of rats Microinjection of cAMP (1.0μg) into RT increased sleep and decreased wakefulness compared with control; microinjection of methylene blue (MB,1.0μg) into RT enhanced sleep and reduced wakefulness compared with control.5. The effects of the 5-HTergic nerve fibers project from DRN to RT on sleep-wakefulness cycle of rats5.1 When L-Glu (0.2μg) was microinjected into DRN and normal sodium (NS,1.0μg) was microinjected into bilateral RT. We found that sleep was decreased and wakefulness was increased compared with control; when L-Glu (0.2μg) was microinjected into DRN and methysergide (MS,1.0μg), a non-selective 5-HT antagonist, was microinjected into bilateral RT, We found that sleep was enhanced and wakefulness was reduced compared with L-Glu group.5.2 When p-chlorophenylalanine (PCPA, 10μg) was microinjected into DRN and NS (1.0μg) was microinjected into bilateral RT, We found that sleep was increased and wakefulness was decreased compared with control; microinjection of 5-hydroxytryptaphan (5-HTP, 1.0μg), which can convert to 5-HT by the enzyme tryptophane hydroxylase and enhance 5-HT into bilateral RT, could block the effect of microinjection of PCPA into DRN on sleep-wakefulness cycle.

本研究采用脑立体定位、核团插管、微量注射、多导睡眠描记等方法，研究丘脑网状核(nucleus reticularis thalami，RT)中γ-氨基丁酸(gamma-amino butyric acid ，GABA)能神经元、一氧化氮(nitrogen monoxidum，NO)能神经元、阿片肽类神经递质、环一磷酸腺苷(cyclic adenosine monophosphate，cAMP)及中缝背核(nucleus raphes dorsalis，DRN)至RT的5-羟色胺(5-hydroxytryptamine，5-HT)能神经纤维投射对大鼠睡眠-觉醒周期的影响及其作用机制。1 RT内GABA能神经元对大鼠睡眠-觉醒周期的影响1.1大鼠RT内微量注射GABA合成关键酶抑制剂3-巯基丙酸(3-MP，5μg)，注射当天睡眠时间略有减少，第二日睡眠时间显著减少，觉醒时间明显增多，第三、四日睡眠和觉醒时间逐渐恢复至正常。1.2大鼠RT内微量注射GABA受体激动剂GABA( 1.0μg)后，与生理盐水组比较，睡眠时间增加，觉醒时间减少；而RT内微量注射L-谷氨酸(glutamic acid， L-Glu， 0.2μg)后，睡眠时间减少，觉醒时间增加；RT内微量注射GABAA受体阻断剂荷包牡丹碱(bicuculline，BIC，1.0μg)后，睡眠时间减少，觉醒时间增加；RT内微量注射GABAB受体激动剂氯苯氨丁酸(baclofen，BAC，1.0μg)后，产生了与GABA相似的促睡眠效果。2 RT内NO能神经元对大鼠睡眠-觉醒周期的影响2.1大鼠RT内微量注射NO的前体L-精氨酸(L-Arg，0.5μg)后，与生理盐水组对比，睡眠时间略有减少，但无显著性意义；而RT内微量注射NO的供体硝普钠(Sodium Nitroprusside，SNP，0.2μg)后可明显增加觉醒时间，缩短睡眠时间；微量注射一氧化氮合酶抑制剂L-硝基精氨酸(L-arginine，L-NNA，2.0μg)后，引起睡眠时间增多，觉醒时间减少。2.2大鼠RT内同时微量注射L-NNA(2.0μg)和SNP(0.2μg)后与L-NNA组比较发现SNP逆转了L-NNA的促睡眠作用；RT内同时微量注射L-NNA(2.0μg)和L-Arg(0.5μg)后，与L-NNA(2.0μg)组比较发现L-Arg可以增加觉醒而缩短睡眠，其促觉醒作用未能被NOS的抑制剂L-NNA所逆转。3 RT内阿片肽对大鼠睡眠-觉醒周期的影响3.1大鼠RT内微量注射硫酸吗啡(morphine sulfate，MOR，1.0μg)后与生理盐水组对比，睡眠时间减少而觉醒时间增加； RT内微量注射阿片肽受体拮抗剂盐酸纳洛酮(naloxone hydrochloride，NAL，1.0μg)后与生理盐水组比较，睡眠时间增加而觉醒时间减少。3.2大鼠RT内同时微量注射MOR(1.0μg)和NAL(1.0μg)后，与生理盐水组对比，原有的MOR促觉醒效果和NAL的促睡眠效果都没有表现。4 RT内环一磷酸腺苷信使对大鼠睡眠-觉醒周期的影响大鼠RT内微量注射cAMP(1.0μg)后与NS(1.0μg)组比较，睡眠时间增多而觉醒时间减少；RT内微量注射亚甲蓝(methylene blue，MB，1.0μg)后，与NS组比较，睡眠时间增多而觉醒时间减少。5中缝背核投射到丘脑网状核的5-羟色胺能神经纤维对大鼠睡眠-觉醒周期的影响5.1大鼠DRN内微量注射L-Glu(0.2μg)，同时在双侧RT内微量注射NS (1.0μg)后，与对照组(DRN和双侧RT注射NS， 0.2μg)比较，睡眠时间减少，觉醒时间增多；大鼠DRN内微量注射L-Glu(0.2μg)，同时在双侧RT内微量注射二甲基麦角新碱(methysergide， MS， 1.0μg )后，与对照组(DRN注射L-Glu 0.2μg，双侧RT注射NS 1.0μg)比较，睡眠时间增多，觉醒时间减少。5.2大鼠DRN内微量注射对氯苯丙氨酸(p-chlorophenylalanine，PCPA，10μg)，同时在双侧RT内微量注射NS (1.0μg)后，与对照组(DRN和双侧RT注射NS， 1.0μg)比较，睡眠时间增多，觉醒时间减少；大鼠DRN内微量注射PCPA(10μg)，产生睡眠增多效应后，在双侧RT内微量注射5-羟色胺酸(5-hydroxytryptaphan ， 5-HTP， 1.0μg )后，与对照组(DRN注射PCPA 10μg，双侧RT注射NS 1.0μg)比较，睡眠时间减少，觉醒时间增多。

Results: Compared with the C group, the contents of CORT in blood and ACTH in pituitary gland are increased at the different time points. Compared with the C group ,the contents of CR11 in Hypothalamus are enhanced in S and SAI groups on list but those are reduced in S and SM groups on 16st and 21st. Compared with the S group ,the contents of CORT in blood and ACH in pituitary gland are increased in SM group on 11 at. Compared with the SM groups, the contents of CORT in blood are reduced in TKXLD groups on 11st, l6st and 21st. Compared with the SAT group respectively, the contents of CRH in Hypothalamus are decreased in TKXLD group on list and those in TKXLD group are increased on 16st and 21st.

结果：实验第11、16和21天悬吊组和悬吊加辐射组大鼠血清CORT和垂体ACTH含量高于正常组；悬吊组和悬吊加辐射组大鼠下丘脑CRH含量在实验第11天高于正常组，在实验第16天和21天低于正常组；实验第11天悬吊加辐射组大鼠血清中CORT的含量高于悬吊组；实验第11、16和21天，中药组大鼠血清CORT含量低于悬吊加辐射组，垂体ACTH含量低于悬吊加辐射组；中药组大鼠下丘脑CRH含量在实验第11天低于悬吊加辐射组，在实验第16天和21天高于悬吊加辐射组。

When aborted rats in group E were treated with the dose of 50 IU/g IFN-γ, expression of TNF-αin this group was compared with the other four groups, the results were as follows: Compared with group A, the nucleus pre-opticus medialis, nucleus pre-opticus mango celluaris, nucleus periventricularis hypothalami, nucleus ventromedialis hypothalami, nucleus dorsomedialis hypothalami, nucleus arcuatus hypothalami and pars intermedia had a highly expression of TNF-α, in the other nuclei of hypothalamus, neurohypophsis and adenohypophysis, the change was not significantly, the expression of TNF-αin follicle and corpus luteum were decreased distinctly, the variation in uterus was slight and no difference was exsited; Compared with group B, the expression of TNF-αin nucleus nucleus lateralis hypothalami, nucleus ventromedialis hypothalami, nucleus arcuatus hypothalami, adenohypophysis and uterus presented a remarkable enhancement, conversely, the TNF-αexpression in the nucleus pre-opticus medialis, nucleus pre-opticus suprachiasmaticus, nucleus supra-opticus, nucleus pre-opticus mango celluaris, nucleus periventricularis hypothalami, nucleus paraventricularis hypothalami, neurohypophsis, follicle and corpus luteum had been significantly decreased; Compared with group C, the nucleus pre-opticus medialis, nucleus ventromedialis hypothalami, nucleus dorsomedialis hypothalami, nucleus arcuatus hypothalami, adenohypophysis, pars intermedia and uterus had upregulated the expression of TNF-α, however, it was decreased in the nucleus periventricularis hypothalami and corpus luteum; When compared with group D, the expression of TNF-αin nucleus lateralis hypothalami, nucleus dorsomedialis hypothalami, nucleus ventromedialis hypothalami, pituitary, follicle and ovary stroma were obviously enhanced, while in the nucleus supra-opticus and nucleus arcuatus hypothalami, the situation was definitely opposite, in the other parts, the change was not significant.

外源腹腔注射50 IU/g IFN-γ后，同A组相比：E组TNF-α免疫阳性产物在下丘脑视前内侧核、视前大细胞核、室周核、腹内侧核、背内侧核、弓状核、垂体中间部显著增高，在下丘脑其它核团、神经垂体、垂体前叶较A组差异不显著，在卵泡、黄体中阳性产物表达均较A组显著降低，在子宫中表达较A组变化不明显；同B组相比：E组TNF-α免疫阳性产物在下丘脑外侧核、腹内侧核、弓状核、垂体前叶、子宫均显著增高，在视前内侧核、视交叉上核、视上核、视前大细胞核、室周核、室旁核、神经垂体、卵泡、黄体中阳性物质表达均显著降低；同C组相比：E组下丘脑视前内侧核、腹内侧核、背内侧核、弓状核、垂体前叶与中间部、子宫均显著增高，在室周核、黄体中均显著降低；同D组相比：E组下丘脑外侧核、背内侧核、腹内侧核、垂体各部、卵泡、卵巢基质均显著增高，在视上核、弓状核显著降低，在其它部位表达变化差异不显著。

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compared with：相比

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compared with：比

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