安定降低家兔血压和减少刺激下丘脑诱发室性期前收缩的机制分析

家兔62只,用乌拉坦(700mg/kg)和氯醛醣(35mg/kg)静脉麻醉,三碘季铵酚制动,在人工呼吸下进行实验。用电刺激下丘脑近中线区的方法诱发室性期前收缩(HVE)。静脉注射安定(0.5mg/kg)可降低基础血压(BP),减弱刺激下丘脑引起升压反应(指收缩压峰值SBP_(max))和减少HVE。在双侧延髓腹外侧头端区(rVLM)微量注射氟安定(200μg溶于0.5μl中),γ-氨基丁酸(GABA)(6μg溶于0.5μl中)均能降低BP、SBP_(max)和减少HVE,若微量注射印防己毒素(7.5μg溶于0.5μl中)则可使BP上升并增多HVE。而于双侧延髓腹外侧尾端区(cVLM)微量注射同样剂量氟安定、GABA则无上述反应。安定降低BP、SBP_(max)和减少HVE的作用可被双侧rVLM区微量注射GABA受体拮抗剂荷包牡丹碱(3μg溶于0.5μl中)或印防己毒素所消除,但在双侧rVLM区微量注射甘氨酸受体拮抗剂士的宁(1μg溶于0.5μl中)、阿片受体拮抗剂纳洛酮(0.5μg溶于0.5μl中)、胆碱能阻断药阿托品(0.25μg溶于0.5μl中)、东莨菪碱(1.5μg溶于0.5μl中)后仍然存在。 上述结果提示,在双侧rVLM应用GABA受体拮抗剂可消除安定降低BP、SBP_(max)和减少HVE的作用,安定降低BP、SBP_(max)和减少HVE的作用可能通过GABA这一中间环节,而胆碱能受体、阿片受体、甘氨酸受体可能不起重要作用。     

急性低氧下中枢加压素对循环活动的效应

为了解中枢加压素是否参与了低氧下循环活动的维持,本实验在同步监测麻醉大鼠血压(BP)、心率(HR)、左、右心室压(LVP,RVP)及其dP/dt的条件下,观察侧脑室注射(icv.)加压素V_1-、V_2-受体拮抗剂(各4μg)及同体积人工脑脊液(ACSF)(8μl)三组不同处理大鼠在低氧下循环指标的变化。于侧脑室注射药物10min后,让大鼠吸入含氧9％的氮氧混合气体15min,然后复氧。实验中观察到三组大鼠在低氧期间,循环活动各指标均下降。但icv.V_1-受体拮抗剂一组大鼠MBP,HR,左心室压峰值(LVSP)、左心室压力变化率最大值(LV+dp/dt_(max))等循环指标下降幅度显著大于对照组(P<0.01);复氧后两组间循环指标差异逐渐消失。常氧下icv,4μg V_1-受体拮抗剂及8μl ACSF对循环活动无影响。常氧下icv.V_2-受体拮抗剂导致大鼠HR,MBP,LVSP,LV+dp/dt_(max)出现一过性升高,但在低氧、复氧期间,该组与对照组循环活动各指标间无显著性差异。上述实验结果提示低氧应激期间,中枢内源性AVP对大鼠循环功能的维持具有重要作用。该效应是通过其V_1-受体介导的,与其V_2-受体无关。在正常情况下,中枢AVP对心血管活动无紧张性影响。     

γ—氨基丁酸在联合皮层抑制C类纤维皮层诱发电位效应中的作用

电刺激猫大脑皮层前外侧回联合区(ALA)对隐神经C类纤维传入引起的体感皮层(SI)诱发电位(C-CEP)有明显的抑制作用;侧脑室注射γ-氨基丁酸(GABA)能使C-CEP的幅值显著变小,潜伏期延长,表明GABA对C-CEP也有抑制作用;侧脑室注射GABA受体拮抗剂荷包牡丹硷后,电刺激ALA对C-CEP的抑制作用明显减弱,提示内源性GABA的释放可能参与大脑皮层联合区对C-CEP的调制过程。     

延髓中线区微量注射5-羟色胺、吗啡等药物对刺激兔下丘脑诱发室性期前收缩的影响

兔廷髓中线区微量注射5-羟色胺(10μg溶于2μl中)或吗啡(10μg溶于2μl中)可减少刺激下丘脑诱发的室性期前收缩(HVE),微量注射肉桂硫胺(4μg溶于2μl中)或纳络酮(4μg溶于2μl中)可在多数动物中引起 HVE 短时内增多。先微量注射肉桂硫胺(2μg溶于1μl中)再微量注射吗啡(10μg溶于1μl中)或先微量注射纳络酮(2μg溶于1μl中)再微量注射5-羟色胺(10μg溶于1μl中),均不能明显影响吗啡、5-羟色胺对 HVE 的抑制效应。延髓中线区微量注射去甲肾卜腺素(2—4μg溶于2—4μl中)不能象在中脑中央灰质微量注射那样使 HVE 增多。对照组动物延髓中线区微量注射生理盐水(2μl)对 HVE 无明显影响。结果提示:延髓中线区5-羟色胺受体和阿片受体的活动对 HVE 有抑制作用。     

延髓腹外侧头端区在第四脑室注射 P物质所致升压中的作用

实验用乌拉坦麻醉、肌内麻痹、人工呼吸的家兔。将Ｐ物质（ＳＰ,０．８ｎｇ／ｋｇ溶于１０μｌ人工脑脊液中）注入第四脑室引起肺动脉压（ＰＡＰ）升高或降低,但对坟反应为主。与此同时,颈劝脉压（ＣＡＰ）上升,心率（ＨＲ）减慢;而在第四及室内注入同容积的人工脑脊液对ＰＡＰ,ＣＡＰ和ＨＲ无明显影响。若在ｉｖｔＳＰ之前,预先向双侧延髓腹外侧头端区微量注射ＳＰ受体拮抗剂－ＳＰ（５＝１０ｎｇ溶于０．５μｌ人工脑脊液中     

大鼠脊髓蛛网膜下腔注射γ-氨基丁酸受体激动剂对降低血压和徐缓心率的作用

给大鼠脊髓蛛网膜下腔注射γ-氨基丁酸(GABA)受体激动剂异鹅羔胺(0.125—0.25μg)能显著降低动物的动脉血压和心率。这种作用可被 GABA 受体阻断剂氯甲基荷包牡丹碱(1.0μg)所翻转,且具有剂量-效应关系。另一种 GABA 受体激动剂 THIP(5μg)以及GABA(500μg)也同样具有降低血压和心率的作用。说明激活脊髓内 GABA 受体具有降低血压和减慢心率的作用。肾上腺素α受体阻断剂育亨宾(30μg)能翻转可乐宁(0.25μg)的降压作用,但不能对抗异鹅羔胺的作用;GABA 受体阻断剂氯甲基荷包牡丹碱可翻转异鹅羔胺的作用,对可乐宁则无效。说明脊髓内异鹅羔胺和可乐宁降血压和减慢心率的作用没有相互的依存关系。     

家兔延髓腹侧区内注射吗啡对刺激下丘脑诱发室性期前收缩的抑制作用

家免63只,用乌拉坦(700mg/kg)和氯醛醣(35mg/kg)静脉麻醉,三碘季铵酚制动,在人工呼吸下进行实验。用电刺激下丘脑近中线区的方法诱发定性期前收缩(HVE)。双侧延髓腹侧区内微量注射吗啡(5μg 溶于0.5—1μl 中)可抑制 HVE。在双侧延髓腹侧区微量注射纳洛酬(2μg 溶于0.5—1μl 中)能阻断电刺激中缝核尾端或中缝核尾端内微量注射 L-谷氨酸钠(50mmol/L,0.5—1μl)对 HVE 的抑制效应,也能减少或消除刺激腓深神经对 HVE 的抑制作用。上述结果提示,延髓腹侧部阿片受体的激活可抑制 HVE;另外,腓深神经传入冲动也可能通过中缝核尾端激活延髓腹侧区的阿片受体而抑制 HVE。     

脑内γ—氨基丁酸（GABA）在急性失血性低血压时对血压的影响

本实验通过大鼠和家兔侧脑室注射GABA受体阻断剂荷包牡丹碱(Bicuculline)研究正常血压和急性失血性低血压时,中枢GABA对血压的调节作用。结果表明:在两种情况下,荷包牡丹碱均有明显的升压作用,但后者显著大于前者,且这种升压效应不被去肾或去肾上腺所影响。同时发现:急性失血性低血压时,兔脑脊液中GABA含量明显增加,提示在急性失血性低血压时,中枢GABA具有阻止血压回升的作用,这种作用是通过对中枢交感神经系统的抑制引起的。     

一叶萩碱是新的GABA受体拮抗剂

近年来国内外学者对一叶荻碱的中枢兴奋作用机制进行了较深入的研究,证明一叶萩碱是一种类似荷包牡丹碱的 GABA 受体拮抗剂。作者等在小鼠实验中,应用无拮抗作用剂量的 GABA(10μg,icv)与苯甲二氮(?)(安定,diazepam,0.18mg/kg(?)in)合并用药(?)对一叶萩碱惊厥有显著拮抗作用。印防己毒素(GABA 激活的氯离子通道阻滞     

兴奋大鼠延髓A1区引起降压、降心率效应的机制

在水合氯醛麻醉、箭毒化、人工呼吸的大鼠,观察到:(1) A_1区注入谷氨酸钠引起明显的血压下降和心率减慢。(2) 切断双侧颈迷走神经明显衰减A_1区的降压,降心率效应。(3) 延髓头端腹外侧区(RVL)预先注射酚妥拉明或心得安,均能明显衰减谷氨酸钠兴奋A_1区的降压效应,A_1区的降心率作用基本不受影响,将纳洛酮注入RVL后,A_1区的降压和降心率效应均无明显变化;注射荷包牡丹碱入RVL则使A_1区的降压、降心率效应反转。(4) RVL内注入酚妥拉明或心得安本身使基础血压降低,注射荷包牡丹碱入RVL则使基础血压升高(提示RVL内的α-,β-受体中介对RVE加压神经元的紧张性兴奋作用,GABA受体中介紧张性抑制作用);另一方面,RVL内注入心得安使基础心率减慢、注入纳洛酮或荷包牡丹碱使基础心率加快(说明β-受体中介紧张性心加速效应,阿片受体和GABA受体中介紧张性心抑制效应)。     

Effects of intracerebroventricular injections of thyrotropin releasing hormone on gastrointestinal propulsive motility in rats

The effect of intracerebroventricular (ICV) injections of thyrotropin releasing hormone (TRH) on the propulsive motility of the gastrointestinal tract was examined in rats. The distance travelled by charcoal meal through the small intestine, measured in terms of percentage of its total length, was recorded as the index of propulsive motility. The results were as follows: (1) The propulsive distance of charcoal meal was significantly reduced in a dose-dependent manner after ICV injections of TRH (1 microgram/10 microliters, 5 micrograms/10 microliters or 10 micrograms/10 microliters) (P less than 0.01-0.001) The effects were abolished by injection of atropine (5 micrograms/10 microliters ICV). (2) The gastrointestinal propulsive motility decreased markedly (P less than 0.01) after injection of a larger dose of TRH (50 micrograms/100 g) into the hypodermis. The effects were not completely blocked by subcutaneous injections of propranolol (5 mg/kg). (3) No effects (P greater than 0.05) were found on the inhibition of gastrointestinal propulsive motility after ICV injections of regitine (2.5 mg/kg im, 50 micrograms/50 microliters ICV) or propranolol (5 mg/kg im, 50 micrograms/50 microliters ICV). The results indicate that TRH has an inhibitory effect on the propulsive motility of gastrointestinal tract, which may be mediated via the non-adrenergic inhibitory nerve of the vagal nerves.     

大鼠尾壳核的GABA能传递在条件性行为调控中的作用

本工作采用了行为和脑内注射相结合的方法研究了大鼠尾壳核的 GABA 能传递在条件性行为调控中的作用。在分辨学习的基础上训练大鼠完成条件性回避任务,以比较药物对分辨学习和条件性回避的不同效应。实验结果表明,于大鼠双侧尾壳核内分别注入 γ-氨基丁酸(GABA)(每侧100μg/μl)和 GABA 受体激动剂蝇蕈醇(Muscimol)(每侧0.1μg/μl)后可暂时抑制条件性回避反应的出现,但分辨学习无明显影响。作为对照,于尾壳核内注入等量的生理盐水则既不影响条件性回避反应,也不影响分辨学习。在条件性回避反应被 Muscimol抑制后于尾壳核内再注入 GABA 受体阻断剂印防己毒素(PTX)(每侧0.1μg/μl)则可拮抗Muscimol 的行为抑制效应,即条件反应的出现率可恢复到或接近注射前水平。实验结果表明,大鼠尾壳核的 GABA 能传递在条件性行为调控中的重要作用。     

GABA/BZ-and NMDA-receptor interaction in digoxin-induced convulsions in rats.

Digoxin (7.5 micrograms icv) induced 'pop-corn' type of convulsions and 100% mortality. The GABA-ergic agents produced varying degree of protection against digoxin-induced neurotoxicity. Diazepam (4 mg/kg) offered significant protection whereas pentobarbital (5 mg/kg) and baclofen (5 mg/kg) markedly reduced per cent mortality, but ethanol (2 g/kg), progabide (50 mg/kg) and muscimol (0.5 mg/kg) as well as GABA (50 mg/kg) could not offer significant protection in doses used. GABA-ergic agonists; GABA, baclofen, diazepam and pentobarbital when administered along with MK-801 (0.5 mg/kg) a non-competitive NMDA antagonist, a potentiation of anticonvulsant action of MK-801 was observed. MK-801 showed potent anticonvulsant profile in dose range (0.25-1 mg/kg) studied. A synergistic influence of Mg2+ and K+ ions on NMDA receptor antagonism was also observed. A role of GABA-ergic facilitation and NMDA antagonism as a potential anticonvulsant approach in digoxin-induced convulsions in rats has been suggested.     

Cortexolone as a modulator of diazepam activity

The influence of ACTH (200 micrograms/kg), corticosterone (20 mg/kg) and cortexolone (20 mg/kg) on the anxiolytic activity of diazepam was studied. ACTH partly and corticosterone completely blocked the action of diazepam. Cortexolone injection 30 min before the administration of diazepam induced a 100% anxiolytic effect of diazepam in the range of doses from 0.1 to 0.3 mg/kg (ED50 of anxiolytic diazepam effect is 0.2 mg/kg). The role of stress hormones in the regulation of psychotropic drug activity is discussed.     

Diazepam Increases γ-Aminobutyric Acid in Human Cerebrospinal Fluid

In 11 neurological patients, levels of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) were determined in cerebrospinal fluid (CSF) before and 1, 3, 5, and 8 min after intravenous injection of diazepam (2 or 5 mg). GABA levels increased progressively after intravenous injection of 5 but not 2 mg of the benzodiazepine, the differences from preinjection values being significant at 3, 5, and 8 min. Furthermore, when relative CSF GABA alterations determined after injection of diazepam were compared to those determined in sequential CSF aliquots of 10 patients without diazepam injection, mean GABA increases after diazepam were significantly different from controls in all CSF fractions. The data suggest that, in addition to its well-known effects on postsynaptic GABA function, diazepam may exert effects on endogenous GABA concentrations and/or on GABA release in the human CNS as reflected by elevation of GABA levels in human CSF.     

Effects of treatment with GABA(A) receptor subunit antisense oligodeoxynucleotides on GABA-stimulated 36Cl- influx in the rat cerebral cortex

GABA(A) receptor function was studied in cerebral cortical vesicles prepared from rats after intracerebroventricular microinjections of antisense oligodeoxynucleotides (aODNs) for alpha1, gamma2, beta1, beta2 subunits. GABA(A) receptor alpha1 subunit aODNs decreased alpha1 subunit mRNA by 59+/-10%. Specific [3H]GABA binding was decreased by alpha1 or beta2 subunit aODNs (to 63+/-3% and 64+/-9%, respectively) but not changed by gamma2 subunit aODNs (94+/-5%). Specific [3H]flunitrazepam binding was increased by alpha1 or beta2 subunit aODNs (122+/-8% and 126+/-11%, respectively) and decreased by gamma2 subunit aODNs (50+/-13%). The "knockdown" of specific subunits of the GABA(A )receptor significantly influenced GABA-stimulated 36Cl- influx. Injection of alpha1 subunit aODNs decreased basal 36Cl- influx and the GABA Emax; enhanced GABA modulation by diazepam; and decreased antagonism of GABA activity by bicuculline. Injection of gamma2 subunit aODNs increased the GABA Emax; reversed the modulatory efficacy of diazepam from enhancement to inhibition of GABA-stimulation; and reduced the antagonist effect of bicuculline. Injection of beta2 subunit aODNs reduced the effect of diazepam whereas treatment with beta1 subunit aODNs had no effect on the drugs studied. Conclusions from our studies are: (1) alpha1 subunits promote, beta2 subunits maintain, and gamma2 subunits suppress GABA stimulation of 36Cl- influx; (2) alpha1 subunits suppress, whereas beta2, and gamma2 subunits promote allosteric modulation by benzodiazepines; (3) diazepam can act as an agonist or inverse agonist depending on the relative composition of the receptor subunits: and (4) the mixed competitive/non-competitive effects of bicuculline result from activity at alpha1 and gamma2 subunits and the lack of activity at beta1 and beta2 subunits.     

Central somatostatinergic regulation of growth hormone secretion in dwarf chickens.

1. Basal circulating growth hormone (GH) concentrations in sex-linked-dwarf (SLD) chickens were unaffected by the intracerebroventricular (icv) injection of 10, 50 or 100 micrograms somatostatin (SRIF). 2. The GH response to systemic thyrotropin-releasing hormone (TRH; 10 micrograms/kg, iv) was, however, 'paradoxically' enhanced 20 min after icv SRIF administration. 3. A lower dose (1.0 micrograms) of SRIF had no effect on basal or TRH-induced GH release. 4. High-titre SRIF antisera (4 microliters) also had no acute effect on basal plasma GH concentrations, but augmented the GH response to TRH challenge. 5. SRIF would appear to act at central sites to modulate stimulated GH secretion in SLD chickens.     

Effect of Withania somnifera Dunal root extract against pentylenetetrazol seizure threshold in mice: possible involvement of GABAergic system

Withania somnifera (ashwagandha) is a widely used herb in the Ayurvedic system of medicine. The objective of the present study was to elucidate the effect of W. somnifera root extract (Ws) alone or in combination with exogenous gamma-amino butyric acid (GABA), a GABA receptor agonist or with diazepam, a GABA receptor modulator against pentylenetetrazol (PTZ, iv) seizure threshold in mice. Minimal dose of PTZ (iv, mg/kg) needed to induce different phases (myoclonic jerks, generalized clonus and tonic extension) of convulsions were recorded as an index of seizure threshold. Ws (100 or 200 mg/kg, po) increased the PTZ seizure threshold for the onset of tonic extension phase whereas a lower dose (50 mg/kg, po) did not show any effect on the seizure threshold. Co-administration of a sub-effective dose of Ws (50 mg/kg, po) with a sub-protective dose of either GABA (25 mg/kg, ip) or diazepam (0.5 mg/kg, ip) increased the seizure threshold. The results suggested that the anticonvulsant effect of W. somnifera against PTZ seizure threshold paradigm involved the GABAAergic modulation.     

Central antihypertensive properties of muscimol and related gamma-aminobutyric acid agonists and the interaction of muscimol with baroreceptor reflexes.

The central antihypertensive properties of four gamma-aminobutyric acid (GABA) analogs were characterized in anesthetized cats with implanted intracerebroventricular cannulae. An intracerebroventricular infusion (icv) of muscimol, 0.1--0.5 microgram/min (total dose: 1--5 micrograms, icv), substantially reduced mean arterial pressure and slightly reduced heart rate. The compound was not hypotensive at 5 micrograms, iv (total dose) and only slightly hypotensive after an intracisternal injection (5 micrograms). Kojic amine (2-aminomethyl-5-hydroxy-4H-pyran-4-one) and baclofen were also hypotensive following an intracerebroventricular infusion, but they were less active than muscimol. GABA, at 15--150 micrograms/min, icv (total dose, 150--1500 micrograms, icv), was not hypotensive by itself and unlike muscimol its activity was not enhanced in cats pretreated with nipecotic acid, an uptake inhibitor of GABA. The ability of muscimol to interfere with baroreceptor reflexes was considered in experiments in which reflex vasoconstrictor (carotid occlusion) and reflex vasodilatation (acute elevation in mean arterial pressure with norepinephrine) was measured in the perfused hindlimb of cats previously prepared with intracerebroventricular cannulae. Muscimol significantly attenuated the response to bilateral carotid occlusion and completely abolished reflex vasodilatation. The results suggest that GABA agonists and analogs may regulate blood pressure centrally and, through an interaction with the central nervous system, may attenuate baroreceptor reflexes.