心钠素在GABA调节下丘脑正中隆起LHRH释放的介导作用研究

已证明脑神经递质GABA可促进下丘脑正中隆起LHRH神经元末梢的释放功能,这一作用可能通过抑制某些抑制性神经递质或调质释放而实现。本研究旨在观察作为体内循环调节肽的心钠素在大鼠脑内的分布,对LHRH释放的影响及对GABA调节LHRH释放的中介作用。实验结果发现:(1)雄性大鼠不同脑区心钠素的含量不同,其中以下丘脑正中隆起组织含量最高;(2)不同浓度的心钠素(10~(-8)-10~(-6)mol/L)可显著抑制LHRH从下丘脑正中隆起释放,并呈现剂量反应关系。(3)GABA(10~(-6)mol/L组)可显著抑制心钠素的释放;GABA的这一抑制作用可被GABA受体拮抗剂荷包牡丹碱(Bicuculline)完全反转。上述结果提示心钠素参与调节下丘脑正中隆起(ME)LHRH神经元末梢的释放机能,它亦可能介导GABA对下丘脑正中隆起LHRH的调节作用。     

虎纹蛙促性腺激素释放激素分泌调节的离体研究

利用离体静态孵育系统和放射免疫测定法,研究了性成熟的虎纹蛙雌蛙离体的视前－下丘脑－正中隆起（P－H－ME）片段促性腺激素释放激素（GnRH)的分泌调节。结果表明：γ－氨基丁酸（GABA）对成熟前期蛙离体P－H－ME片段的哺乳类GnRH（mGnRH)的释放有显著的刺激作用;随着GABA作用浓度的增加,刺激作用逐渐增强。100μmol/L的多巴胺（DA）及1μmol/L和10μmol/L的雌二醇(E2）则显著抑制鸡ⅡGnRH(cGnRH-Ⅱ)的释放。10μmol/L和100μmol/L的睾酮（T）以及10μmol/L的E2显著刺激冬眠期蛙P－H－ME片段mGnRH的释放。这些结果表明,GABA,DA及E2和T对虎蚊蛙GnRH的释放有直接的调节作用。     

家兔延髓腹外侧区内注射荷包牡丹碱对安定降低心脏功能的影响

家兔48只,用乌拉坦(1g/kg)静脉麻醉,三碘季铵酚制动,在人工呼吸下进行实验。静脉注射安定(0.5mg/kg)以及侧脑室注射氟安定(2mg溶于50μl人工脑脊液中)或γ-氨基丁酸(GABA)(300μg溶于50μl人工脑脊液中)都可降低心室内压峰值(PLVP),减少心力环面积(ACFL)和心室内压最大上升速率(dp/dt_(max))。安定和氟安定对PLVP、ACFL和dp/dt_(max)的抑制作用可被预先在侧脑室注射GABA受体拮抗剂印防己毒素(15μg溶于50μl人工脑脊液中)或双侧延髓腹外侧头端区(γVLM)微量注射GABA受体拮抗剂荷包牡丹碱(3μg溶于0.5μl生理盐水)所阻断。而在延髓网状巨细胞核中间部、网状小细胞核、孤束核或面神经核内注射同样剂量的荷包牡丹碱则不能阻断。 这些结果提示:延髓腹侧部GABA受体的激活可抑制心脏功能,安定对心脏功能的抑制作用可能通过激活延髓腹侧区GABA受体而实现。     

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。     

霉菌毒素 Dihydroxyaflavinine 非竞争性抑制在爪蟾卵母细胞内表达的 GABA_A 受体通道

Dihydroxyaflavinine 是黄曲霉菌(Aspergillus flavus)的吲哚类代谢物。我们将鸡脑mRNA 注入爪蟾卵母细胞表达得到 GABA_A 受体通道,然后用电压箝记录方法定量研究了dihydroxyaflavinine 对 GABA 电流反应的作用。Dihydroxyaflavinine 非竞争性阻断GABA 电流反应(K_I=12μmol/L),撤药后反应迅速恢复。作为比较,青霉素对 GABA_A受体的抑制作用随 GABA 浓度的升高而增加。浓度高达1μmol/L 苯二氮(艹卓)位点配体 Ro 15-1788(KD=0.6—2nmol/L)不能阻断10μmol/L dihydroxyaflavinine 的作用,说明 dihy-droxyaflavinine 不作用于 GABA_A 受体的苯二氮(艹卓)位点。Dihydroxyaflavinine 类似印防已毒素,表观上加速 GABA_A 受体的脱敏过程,而青霉素和荷包牡丹碱与此相反。     

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。     

GABA能神经功能有了新发现

美国耶鲁大学S.A.Tomiko等最近以电生理实验表明,GABA通过荷包牡丹碱(bicuculline,BC)-阻滞性受体,直接作用分离出的大鼠促黑激素(MSH)细胞,结果Cl~-电导增加,膜电位朝Cl~-平衡电位的方向发展,引起静息部位去极化或使高K~ 诱发的去极化难以进行。作者发现GABA首先刺激MSH释放,尔后抑制之,同时抑制K~ 诱发的MSH分泌。作者提供的药理学证据提示,上述分泌和电生理活动所涉及的受体相同。GABA能神经直接作用MSH细胞,改变其释放量,并受GABA产生的电生理特性的变化而影响其功能,这是GABA能神经功能的新发现。多采用大鼠为实验对象,先分离垂体后叶中间部细胞,体外短期培养,并置于柱形器皿内,持续灌流并测定MSH释放量。     

肾上腺髓质素对大鼠损伤性心肌肌浆网功能的改善

通过观察下述五个指标,评价肾上腺髓质素(adrenomedullin,Adm)对大鼠损伤性心肌肌浆网功能的改善程度左心室压力最大变化速率(±dp/dtmax)、肌浆网钙摄取和释放及钙泵活性.皮下注射异丙肾上腺素(isoproterenol,ISO,69μmol/kg体重)制备大鼠心肌损伤坏死模型.摘取心脏后用Adm灌流,观察左心室压力最大变化速率(±dp/dtmax);制备并提纯心肌肌浆网(sarcoplasmicreticulum,SR)膜,测定SRCa2+摄取和释放速率、SR钙泵活性和钙通道蛋白～3H-ryanodine受体的最大结合量.结果发现,5×10-5mol/LAdm灌流能使ISO损伤的大鼠心脏左室±dp/dtmax分别增加16.9%(2?135±281vs1?980±302)和29.2%(1?375±267vs1?064±355,均P<0.05);SRCa2+摄取和释放率分别增加23.0%(15.0±1.4vs12.2±1.2)和43.5%(6.6±1.0vs4.6±0.6,均P<0.01);SRCa2+-ATPase活性和～3H-ryanodine受体最大结合量(Bmax)分别增加24.2%(P<0.01)和42.2%(P<0.05).提示Adm对ISO诱导的大鼠心肌损伤具有保护作用,其机制可能与Adm增加SRCa2+-ATPase活性、增加～3H-ryanodine所致SRCa2+摄取和释放升高有关.外源性给予Adm对损伤心肌可能具有临床治疗作用.     

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。     

γ-氨基丁酸、荷包牡丹碱和L-谷氨酸钠对猫和家兔同侧和对侧图形视觉诱发电位的影响

在猫和家兔大脑半球一侧视区17/18交界处施加γ—氨基丁酸(GABA)、荷包牡丹碱和L—谷氨酸钠,以及用氯化钾和冷冻阻遏的方法,记录对侧和同侧皮层相应处图形视觉诱发电位(PVEP)的变化。讨论了GABA、荷包牡丹碱和L—谷氨酸钠对猫和兔的对侧和同侧PVEP的影响。     

荷包牡丹碱对垂体前叶组织块释放促肾上腺皮质激素的刺激效应

本实验利用垂体组织块离体灌流技术,观察到－氨基丁酸Ａ受体拮抗剂荷包牡丹碱对切除双侧肾上腺９６ｈ后的大鼠垂体前叶ＡＣＴＨ的分泌具有强烈的刺激作用。但同样浓度的荷包牡丹碱对分离的垂体前叶细胞的ＡＣＴＨ分泌无影响。提示肾上腺切除后,－氨基丁酸在垂体前叶直接或通过间接途径抑制ＡＣＴＨ分泌。     

葆包牡丹碱对垂体前叶组织块释放促肾上腺皮质激素的刺激效应

本实验利用垂体组织块离体灌流技术,观察到γ－氨基丁酸Ａ受体拮抗剂荷包牡丹笃切除双侧肾上腺９６ｈ后的大鼠垂体前叶ＡＣＴＨ的分泌具有强烈的刺激作用。但同样浓度的荷包牡丹笃分离的垂体前叶细胞的ＡＣＴＨ分泌无影响,提示肾上腺切除后,γ－氨基丁酸在垂体前叶直接或通过间接途径抑制ＡＣＴＨ分泌。     

Effects of noradrenaline administration on the interrenal gland of the newt, Triturus carnifex: evidence of intra-adrenal paracrine interactions

The existence of paracrine control of steroidogenic activity by adrenochromaffin cells in Triturus carnifex was investigated by in vivo noradrenaline (NA) administration. The effects were evaluated by examination of the ultrastructural morphological and morphometrical features of the tissues as well as the serum levels of aldosterone, NA, and adrenaline (A). In March and July, NA administration increased aldosterone release (from 187.23 +/- 2.93 pg/ml to 878.31 +/- 6.13 pg/ml in March; from 314.60 +/- 1.34 pg/ml to 622.51 +/- 2.65 pg/ml in July) from steroidogenic cells. The cells showed clear signs of stimulation, as evidenced by a strong reduction of lipid content. Moreover, NA administration decreased the mean total number of secretory vesicles in the chromaffin cells in March (from 7.24 +/- 0.18 granules/micro2 to 5.57 +/- 1.88 granules/micro2) and July (from 7.74 +/- 0.74 granules/micro2 to 6.04 +/- 1.13 granules/micro2). In March, however, when T. carnifex chromaffin cells contain both catecholamines, NA (3.88 +/- 0.13 granules/micro2) and A (3.36 +/- 0.05 granules/micro2) in almost equal quantities, NA administration reduced A content (1.29 +/- 1.04 granules/micro2) in the chromaffin cells, enhancing adrenaline secretion (from 681.27 +/- 1.83 pg/ml to 1527.02 +/- 2.11 pg/ml). In July, when the chromaffin cells contain almost exclusively NA granules (NA: 7.42 +/- 0.86 granules/micro2; A: 0.32 +/- 0.13 granules/micro2), NA administration reduced the number of NA granules (5.45 +/- 1.10 granules/micro2), thereby increasing noradrenaline release from the chromaffin cells (from 640.19 +/- 1.65 pg/ml to 1217.0 +/- 1.14 pg/ml). The results of this study indicate that NA influences the steroidogenic cells, eliciting aldosterone release. Noradrenalin effects on the chromaffin cells, increase of NA or A secretion, according to the period of chromaffin cell functional cycle, may be direct and/or mediated through the steroidogenic cells. The existence of intra-adrenal paracrine interactions in T. carnifex is discussed.     

Involvement of central GABAergic neurons in basal and diurnal changes of tuberoinfundibular dopaminergic neuronal activity and prolactin secretion

Central administration of gamma-aminobutyric acid (GABA) has been shown to stimulate the secretion of prolactin (PRL). Whether GABA acts via dopamine, the major PRL-inhibiting hormone, and which GABA receptor type(s) is involved have not been ascertained. Both GABA(A) and GABA(B) receptor agonists and/or antagonists were administered centrally in this study and their effects on both basal and diurnal changes of tuberoinfundibular dopaminergic (TIDA) neuronal activity were determined by measuring the concentration of 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence (ME). Serum PRL level was determined by RIA. Ovariectomized, estrogen-primed Sprague-Dawley rats implanted with intracerebroventricular (icv) cannulae were used. Muscimol (1 ng/3 microl/rat, icv), a GABA(A) receptor agonist, but not baclofen (1-100 ng/3 microl/rat, icv), a GABA(B) receptor agonist, injected in the morning significantly lowered and elevated ME DOPAC and serum PRL levels, respectively at 15 and 30 min. Lower and higher doses of muscimol were not effective. The effects of muscimol could also be prevented by co-administration of bicuculline (0.1-10 ng/3 microl, icv), a GABA(A) receptor antagonist. When bicuculline (10-500 ng/3 microl, icv) was given in the afternoon (at 1500 h), it significantly reversed the lowered ME DOPAC level in the afternoon and prevented the concurrent PRL surge. We conclude that endogenous GABA acting through GABA(A) receptors may play a significant role in the control of basal and diurnal changes of TIDA neuronal activity, and in turn, PRL secretion.     

Release of aldosterone and catecholamines from the interrenal gland of Triturus carnifex in response to adrenocorticotropic hormone (ACTH) administration

The influence of adrenocorticotropic hormone (ACTH) on the interrenal gland of Triturus carnifex was investigated by in vivo administration of synthetic ACTH. The effects were evaluated by examination of the ultrastructural morphological and morphometrical features of the tissues as well as the circulating serum levels of aldosterone, noradrenaline (NA), and adrenaline (A). In June and November, ACTH administration increased aldosterone release (from 281.50 +/- 1.60 pg/ml in carrier-injected newts to 597.02 +/- 3.35 pg/ml in June; from 187.45 +/- 1.34 pg/ml in carrier-injected animals to 651.00 +/- 3.61 pg/ml in November). The steroidogenic cells showed clear signs of stimulation, together with a reduction of lipid content in June and an increase of lipid content in November. Moreover, ACTH administration decreased the mean total number of secretory vesicles in the chromaffin cells in June (from 7.73 +/- 0.60 granules/microm2 in carrier-injected animals to 5.91 +/- 0.40 granules/microm2) and November (from 7.78 +/- 0.75 granules/microm2 in carrier-injected newts to 4.87 +/- 0.40 granules/microm2). In June, however, when T. carnifex chromaffin cells contain almost exclusively NA granules (NA: 7.42 +/- 0.86 granules/microm2; A: 0.32 +/- 0.13 granules/microm2), ACTH decreased NA content (5.52 +/- 0.32 granules/microm2) increasing NA release (from 639.82 +/- 3.30 pg/ml in carrier-injected to 880.55 +/- 4.52 pg/ml). In November, when both catecholamines, NA (3.92 +/- 0.34 granules/microm2) and A (3.84 +/- 0.33 granules/microm2), are present in the chromaffin cells, ACTH administration reduced A content (1.02 +/- 0.20 granules/microm2), enhancing adrenaline secretion (from 681.30 +/- 3.62 pg/ml in carrier-injected newts to 1,335.73 +/- 9.03 pg/ml). The results of this study indicate that ACTH influences the steroidogenic tissue, eliciting aldosterone release. The effects on the chromaffin tissue, increase of NA or A secretion, according to the period of chromaffin cell functional cycle, may be direct and/or mediated through the increase of aldosterone release. Finally, the lack of an increase of A content in the chromaffin cells, or A serum level, following ACTH administration in June might suggest an independence of PNMT enzyme on corticosteroids.     

Effects of adrenaline administration on the interrenal gland of the newt, Triturus carnifex: evidence of intraadrenal paracrine interactions

The existence of paracrine control of steroidogenic activity by adrenochromaffin cells in Triturus carnifex was investigated by in vivo adrenaline (A) administration. The effects were evaluated by examination of the ultrastructural morphological and morphometrical features of the tissues as well as the serum levels of aldosterone, noradrenaline (NA), and adrenaline. In March and July, adrenaline administration reduced aldosterone release (from 187.23 +/- 2.93 pg/ml to 32.28 +/- 1.85 pg/ml in March; from 314.60 +/- 1.34 pg/ml to 87.51 +/- 2.57 pg/ml in July) from steroidogenic cells. The cells showed clear signs of lowered activity: they appeared full of lipid, forming large droplets. Moreover, adrenaline administration decreased the mean total number of secretory granules in the chromaffin cells in July (from 7.74 +/- 0.74 granules/microm(2) to 5.14 +/- 1.55 granules/microm(2)). In this period T. carnifex chromaffin cells contain almost exclusively NA granules (NA: 7.42 +/- 0.86 granules/microm(2); A: 0.32 +/- 0.13 granules/microm(2)). Adrenaline administration reduced noradrenaline content (4.36 +/- 1.40 granules/microm(2)) in the chromaffin cells, enhancing noradrenaline secretion (from 640.19 +/- 1.65 pg/ml to 1030.16 +/- 3.03 pg/ml). In March, adrenaline administration did not affect the mean total number of secretory vesicles (from 7.24 +/- 0.18 granules/microm(2) to 7.25 +/- 1.97 granules/microm(2)). In this period the chromaffin cells contain both catecholamines, noradrenaline (3.88 +/- 0.13 granules/microm(2)), and adrenaline (3.36 +/- 0.05 granules/microm(2)), in almost equal quantities; adrenaline administration reduced adrenaline content (1.74 +/- 0.84 granules/microm(2)), increasing adrenaline release (from 681.27 +/- 1.83 pg/ml to 951.77 +/- 4.11 pg/ml). The results of this study indicate that adrenaline influences the steroidogenic cells, inhibiting aldosterone release. Adrenaline effects on the chromaffin cells (increase of noradrenaline or adrenaline secretion) vary according to the period of chromaffin cell functional cycle. The existence of intraadrenal paracrine interactions in T. carnifex is discussed.     

非洲爪蟾卵母细胞GABAB和GABAc受体介导的电流反应

实验应用双电极电压箝技术,在具有滤泡膜的非洲爪蟾(Xenopuslaevis)卵母细胞上记录到γ-氨基丁酸(γ-aminobutyricacid,GABA)-激活电流。此GABA-激活电流的特点及有关GABA受体类型的研究和分析如下(1)在35.5%(55/155)的受检细胞外加GABA可引起一慢的浓度依赖性的外向电流。(2)GABAA受体的选择性拮抗剂bicuculline(10     

强啡肽抑制离体血管收缩效应的机制

用离体血管电场刺激收缩模型观察到强啡肽明显抑制电场刺激引起的兔耳中心动脉及兔肠系膜上动脉的收缩效应,且呈剂量反应关系,而对股动脉的电场刺激收缩反应无明显影响,强啡肽抑制血管收缩达50％时的用量(IC_(50)值)分别为8.5±1.2×10~(-6)mol/L、5.02±1.3×10~(-7)mol/L及>10~(-6)mol/L。 用药物分析法看到,酚妥拉明(10~(-6)mol/L)可取消电场刺激及去甲肾上腺素引起的血管收缩作用,而强啡肽仅抑制电场刺激致血管收缩作用。 用HPLC法测定孵育液中去甲肾上腺素的含量变化时看到,应用强啡肽(5×10~(-7)mol/L)后孵育液中去甲肾上腺素的含量从对照组的340.56±73.13pg/ml下降至67.91±10.26pg/ml,两组差别有极显著意义(P<0.01)。纳洛酮(10~(-6)mol/L)可完全拮抗强啡肽的这一抑制效应。 以上结果提示强啡肽可能通过抑制交感神经末梢释放去甲肾上腺素,从而产生抑制血管的收缩作用。     

红藻氨酸和GABA受体在中华大蟾蜍卵母细胞的表达

本文报道了利用中华大蟾蜍卵母细胞作为外源性膜蛋白的表达及其功能特性研究的模式系统。将大鼠脑的mRNA微量注入蟾蜍卵母细胞(每个卵母细胞注射50ng),在19℃下经48h以上培养后,由外源mRNA表达的大鼠脑的红藻氨酸和γ-氨基丁酸受体被整合到了卵母细胞膜上。红藻氨酸(5×10~(-5)mol/L)和γ-氨基丁酸(10~(-4)mol/L)所诱导的膜电流分别达到294.0±6.4nA(n=5)和309.5±4.9nA(n=4)。红藻氨酸浓度在10~(-3)mol/L时,其诱导的膜电流达最大值。进而,注射mRNA的卵母细胞,~(36)Cl~-流入速度比对照组高一倍多。这些结果表明,中华大蟾蜍卵母细胞,如同爪蟾卵母细胞一样,能表达具有功能的外源膜蛋白(受体蛋白和离子运输蛋白)。