青藤碱对偏头痛模型大鼠脑干c-fos、c-jun表达的影响

目的 观察青藤碱对偏头痛模型大鼠脑于c-fos、c-jun表达的影响,了解青藤碱有无治疗偏头痛的作用.方法 60只健康Wistar大鼠随机分为空白对照组、模型组、舒马普坦对照组和青藤碱低、中、高剂量治疗组.皮下注射硝酸甘油(NTG)复制实验性偏头痛动物模型,药物干预4h后断头取脑.免疫组化法检测脑干c-fos、c-jun表达水平,显微镜下计数阳性细胞数.结果 与空白对照组相比较,模型组脑干c-fos、c-jun表达明显增多,差异有显著统计学意义(P＜0.01)；与模型组相比较,青藤碱高、中、低剂量组和舒马普坦组脑干c-fos、c-jun表达明显减少,差异有显著统计学意义(P＜0.01)；与舒马普坦组相比较,青藤碱高剂量组脑干c-fos、c-jun表达无差异,无统计学意义(P＞0.05).结论 青藤碱可能通过某些作用机制治疗偏头痛,阻断疼痛刺激信号传入脑干,抑制脑干c-fos、c-jun表达,减轻颅内疼痛长时程反应.     

天舒胶囊对偏头痛动物模型血浆一氧化氮、一氧化氮合酶、降钙素基因相关肽含量及血流动力学的影响

目的 研究天舒胶囊对偏头痛动物模型血浆及脑组织血管活性物质及血流动力学的影响.方法 皮下注射硝酸甘油分别制作大鼠和兔偏头痛模型.给予天舒胶囊后用放免法和分光光度法测大鼠血浆一氧化氮(NO)和一氧化氮合酶(NOS)、降钙素基因相关肽(CGRP)含量;通过免疫组织化学染色观察三叉神经脊束核神经元型NOS(NOS1)和CGRP表达;用经颅多普勒检测兔颈内动脉血流速度改变.结果 模型组大鼠血浆NO、NOS和CGRP较对照组明显升高;经不同剂量天舒胶囊灌胃后大鼠血浆NO、NOS和CGRP的增加受到抑制,尤以中、高剂量组明显(P<0.05～0.01).模型组兔颈内动脉收缩期峰值流速明显下降,经中剂量天舒胶囊干预后流速下降也受到抑制 (P<0.05).免疫组织化学染色发现灌胃天舒胶囊后,偏头痛大鼠三叉神经脊束核NOS1和CGRP表达增加的程度减小(均P<0.05).结论 天舒胶囊可改善偏头痛发作时血管活性物质和神经递质水平失常,从而缓解偏头痛症状.     

天麻制剂通过腺苷途径治疗偏头痛相关的分子机制研究

目的探讨天麻制剂各有效成分对偏头痛模型大鼠的降钙素基因相关肽(calcitonin gene-related peptide,CGRP)与腺苷A1受体(adenosine A1 receptor,A1R)表达的影响。方法将SPF级雄性SD大鼠84只随机分为7组(n=12):假手术组(A组/阴性对照组)、电刺激三叉神经节(electrical stimulation of the trigeminal ganglion,ESTG)模型组(B组)、舒马普坦干预组(C组/阳性对照组)、天麻素干预组(D组)、对羟基苯甲醇干预组(E组)、香英兰醇干预组(F组)、β-谷甾醇干预组(G组)。通过建立ESTG模型,采用酶联免疫吸附测定(enzyme-linked immuno sorbent assay,Elisa)、免疫荧光及Western-Blot技术检测天麻制剂各有效成分对CGRP与A1R表达的影响。结果与A组相比,B组大鼠三叉神经节(trigeminal ganglia,TG)、三叉神经脊束尾核(trigeminal nucleus caudalis,TNC)中的CGRP表达明显增高,A1R的表达明显降低,差异具有统计学意义(P 0. 01)。与B组相比,C、D组大鼠TG、TNC中的CGRP表达明显降低,A1R的表达明显增高,差异具有统计学意义(P 0. 01);而E、F、G组与B组之间差异无统计学意义(P0. 05)。与C组相比,D组大鼠TG、TNC中的CGRP、A1R的表达无明显差异(P 0. 05)。结论预防应用天麻素,与舒马普坦一样可对偏头痛发作起到一定保护作用,而香英兰醇、对羟基苯甲醇及β-谷甾醇对缓解偏头痛的作用疗效甚微,且天麻制剂有效成分中天麻素可通过激活A1R及抑制CGRP表达来抑制偏头痛的发生。     

天舒胶囊对偏头痛大鼠血浆β内啡肽、五羟色胺含量及其脑组织c-fos表达的影响

目的研究天舒胶囊对偏头痛大鼠血浆β内啡肽(β-EP)、五羟色胺(5-HT)含量及其脑组织c-fos表达的影响。方法皮下注射硝酸甘油建立偏头痛大鼠模型,给予天舒胶囊高、中、低剂量灌胃(剂量为7.5g/kg、3.75g/kg和1.88g/kg),按治疗和预防两种方式给药。空白对照组和模型组予以相应剂量的生理盐水灌胃。采用放免法和高效液相色谱法分别测定大鼠静脉血中β-EP、5-HT含量;免疫组化染色观察中脑导水管周围灰质β-EP、5-HT和c-fos阳性表达。结果与模型组比较,天舒胶囊高剂量治疗和预防组、中剂量预防组血浆β-EP水平明显升高(均P<0.05),天舒胶囊高、中、低剂量治疗和预防组血浆5-HT水平明显降低(均P<0.05);天舒胶囊中剂量治疗和预防组中脑导水管周围灰质c-fos表达明显降低(均P<0.05),而β-EP、5-HT表达增高(均P<0.05)。结论天舒胶囊可改善偏头痛发作时血管活性物质和神经递质水平失常,从而改善偏头痛症状。     

舒马普坦通过细胞外信号调节激酶1/2和c-Jun氨基末端激酶信号通路下调三叉神经节内降钙素基因相关肽的表达

目的 探讨舒马普坦对大鼠三叉神经节(TG)离体培养后降钙素基因相关肽(CGRP)表达水平的影响.方法 采用TG离体培养模型,按数字随机表法将54个TG随机分为新鲜组(6个)、对照组(6个)和实验组(7个亚组,每亚组6个,共42个).实验组TG培养液中分别加入4种不同浓度舒马普坦,细胞外信号调节激酶1/2 (ERK1/2)信号通路阻滞剂U0126和PD98059,c-Jun氨基末端激酶(JNK)信号通路阻滞剂SP600125,孵育24h后免疫组织化学染色检测CGRP免疫反应(CGRP-ir)阳性细胞表达,实时定量PCR检测CGRP-mRNA表达量,Western blot定量磷酸化ERK1/2( pERK1/2)和JNK (pJNK)蛋白水平.结果 离体培养24h后,TG内CGRP-ir(+)细胞表达明显增高,0.1和0.5 mg/ml舒马普坦组CGRP-ir(+)细胞百分比、阳性面积、累积吸光度、平均吸光度、CGRP-mRNA水平较对照组明显下降(tPCP=8.652、26.382,tares=6.220、13.917,tIA=5.606、15.904,tM14=2.661、21.748,tmRNA=8.032、15.675,均P＜0.05);而0.02和2.50 mg/ml舒马普坦与对照组CGRP表达差异无统计学意义.Western blot结果显示:0.50 mg/ml浓度舒马普坦显著降低TG内pERKl/2、pJNK水平,降低程度分别接近于10μmol/L的U0126、PD98059和SP600125.结论 一定浓度舒马普坦通过细胞内ERKl/2、JNK信号通路下调大鼠TG离体培养后CGRP的过度表达.     

通心络胶囊对实验性偏头痛大鼠CGRP及α-CGRP基因表达的影响

目的研究通心络胶囊对硝酸甘油性偏头痛大鼠模型血浆降钙素基因相关肽(CGRP)、脑干及三叉神经节αCGRP基因mRNA表达的影响,以探讨通心络治疗偏头痛的可能机制。方法50只SD大鼠随机分为空白对照组、生理盐水组、模型组、苯噻啶防治组、通心络防治组。模型组和药物干预组用硝酸甘油10mg/kg皮下注射造模,每周1次,连续4周,第2次造模后每天分别灌服通心络1.5g/kg/d(通心络组)或苯噻啶0.5mg/kg/d(苯噻啶组)进行干预。第4次造模后用放射免疫法测定血浆CGRP、RTPCR技术测定脑干及三叉神经节区αCGRP基因mRNA的表达。结果与空白对照组比较,反复硝酸甘油造模后,通心络组血浆CGRP水平明显升高(55.63±9.01ng/Lvs45.23±1.79ng/L,P<0.05);苯噻啶组和通心络组αCGRP基因mRNA表达分别为0.67±0.12和0.64±0.12,与空白对照组的0.88±0.09比较,明显下调(P<0.05)。结论通心络胶囊下调了脑干及三叉神经节αCGRP基因mRNA的表达,增加了血浆CGRP含量,提示通心络胶囊不是通过下调血浆CGRP含量而起到预防偏头痛的作用。     

正清风痛宁对偏头痛大鼠脑干IL-1β、TNF-α表达的影响

目的观察正清风痛宁对炎性细胞因子白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)在偏头痛大鼠脑干表达的影响。方法将60只Wistar大鼠(雌雄各半)随机分为空白对照组(空白组)、偏头痛模型组(模型组)、舒马普坦组及正清风痛宁高、中、低剂量干预组(高、中、低剂量组)共6组。除空白组外,其余5组大鼠均建立三酰甘油偏头痛模型,用免疫组化SP法测定各组大鼠脑干IL-1β、TNF-α阳性细胞数。结果 6组大鼠间IL-1β、TNF-α阳性细胞数比较差异有统计学意义(分别F=7.063,P=0.001;F=8.257,P=0.000);组间两两比较,模型组脑干IL-1β(18.9±8.17)、TNF-α(14.30±6.41)阳性细胞数较空白组(5.90±2.69、5.00±1.63)增多(分别t=-4.780、P=0.000,t=-4.444,P=0.000),舒马普坦组(2.80±2.15,t=6.026,P=0.000;0.00±0.00,t=7.052,P=0.000)、中剂量组(7.70±4.76,t=3.745,P=0.000;6.20±1.99,t=3.815,P=0.000)、高剂量组(7.80±5.90,t=3.482,P=0.003;5.90±2.88,t=3.778,P=0.001)IL-1β、TNF-α阳性细胞数均较模型组减少;低剂量组脑干IL-1β(13.5±4.30,t=1.849,P=0.081)、TNF-α(11.30±6.11,t=1.071,P=0.298)阳性细胞数与模型组比较差异均无统计学意义;与舒马普坦组比较,低剂量组(t=-7.037,P=0.000;t=-5.847,P=0.000)、中剂量组(t=-2.966,P=0.011;t=-9.858,P=0.000)、高剂量组(t=-2.517,P=0.022;t=-6.467,P=0.000)脑干IL-1β、TNF-α阳性细胞数均增多;与低剂量组比较,中(t=-2.858,P=0.011;t=-2.510,P=0.022)、高剂量组(t=-2.468,P=0.024;t=-2.527,P=0.021)脑干IL-1β、TNF-α阳性细胞数减少;中、高剂量组脑干IL-1β、TNF-α阳性细胞数差异无统计学意义(t=0.042,P=0.967;t=-0.271,P=0.790)。结论正清风痛宁可能抑制偏头痛大鼠炎性细胞因子IL-1β、TNF-α的表达。     

电刺激三叉神经节诱导偏头痛大鼠脑膜非特异炎症

目的 电刺激三叉神经节建立偏头痛大鼠模型,观察偏头痛大鼠硬脑膜的非特异炎症及5-HTIB/1D受体、降钙素基因相关肽物质(CGRP)在硬脑膜上的分布.方法 采用体重250～300克的雄性SD大鼠,单侧电刺激大鼠三叉神经节建立偏头痛大鼠模型,取大鼠硬脑膜,(1)行HE染色观察非特异炎症;(2)行免疫荧光法检测硬脑膜上5-HT1B/1D受体和CGRP的表达.结果 偏头痛大鼠模型成功建立得到两方面证实:(1)刺激侧大鼠咀嚼肌收缩,口鼻分泌物增多;(2)硬脑膜HE染色显示刺激侧出现炎症反应.初步完成5-HT1B/1D受体及CGRP在硬脑膜上的定位.结论 电刺激三叉神经节可诱导偏头痛大鼠脑膜非特异炎症,5-HT1B/1D受体和CGRP可能在偏头痛疼痛的产生和维持中起着重要的作用.     

瞬时受体电位离子通道A1对慢性偏头痛大鼠模型的作用及氟桂利嗪对其影响

目的本实验通过炎性汤(IS)反复刺激大鼠上矢状窦区硬脑膜疼痛感受器建立慢性偏头痛(CM)大鼠模型,研究瞬时受体电位离子通道A1(TRPA1)及降钙素基因相关肽(CGRP)在CM大鼠模型中的作用,探讨TRPA1在CM发生发展中的可能作用及氟桂利嗪对其影响。方法清洁级SD雄性大鼠48只,体重250~300 g,按随机数字法分为4组(n=12):正常对照组(A组)、假手术组(B组)、CM模型组(C组)及药物干预组(D组)。注射试剂1 h后安静环境中进行大鼠行为学观察及机械刺激缩足反应阈值(PWMT)测定。采用Elisa、Real-Time PCR及Western-Blot技术检测大鼠硬脑膜、三叉神经节(TG)及三叉神经脊束核尾核(TNC)组织部位TRPA1及CGRP表达情况。结果与A组、B组相比较,C组大鼠行为学评分明显升高,PWMT测定值降低,大鼠硬脑膜、TG及TNC组织部位中CGRP及TRPA1表达量均明显上调,差异有统计学意义(P0.05);与C组相比较,D组大鼠行为学评分降低,PWMT测定值升高,硬脑膜、TG及TNC组织部位CGRP及TRPA1表达量均下调,差异有统计学意义(P0.05)。结论 I TRPA1受体可能参与CM发作的病理生理过程;氟桂利嗪可能通过影响TRPA1受体表达,引起CGRP表达下调,从而缓解CM症状。     

帕罗西汀对束缚应激与额叶皮质相关性的影响研究

目的：观察束缚应激对大鼠额叶皮质的影响,探讨帕罗西汀在防治束缚应激引起焦虑的作用机制。方法：复制大鼠束缚应激模型,分为束缚应激组、治疗组和保护组,另设对照组（不束缚应激）,每组均n=10。观察大鼠行为学改变、额叶皮质c-fos基因表达、5-羟色胺（5-HT）和血浆皮质酮含量的变化,观察帕罗西汀干预的影响。结果：与对照组比较,束缚应激组大鼠额叶皮质的c-fos表达显著增加,5-HT表达明显减少,血浆皮质酮含量明显升高;与束缚应激组比较,治疗组和保护组血浆皮质酮含量和额叶皮质c-fos表达明显减少,5-HT表达明显增加。结论：额叶皮质参与了束缚应激反应;帕罗西汀抑制额叶皮质的c-fos基因表达、减少血浆皮质酮含量和增加额叶皮质5-HT含量可能是其抗焦虑作用机制之一。     

Characteristics of the sympathetic innervation of the nictitating membrane and of the vasculature of the nose and tongue of the cat

Summary Vasomotor responses from the nasal mucosa and tongue, and contractions of the nictitating membrane, were recorded on stimulation of the cervical sympathetic or internal carotid nerves.Preganglionic sympathetic nerve fibres which elicited a membrane response possessed a lower threshold than those which evoked nasal vasoconstriction, while the latter displayed a lower threshold than fibres which evoked tongue vasoconstriction. The sympathetic vasodilator fibres to the tongue, whose activity was revealed after-receptor blockade, had a similar threshold to the vasoconstrictor fibres.Membrane contraction, nasal vasoconstriction and occasionally tongue vasoconstriction could be evoked by stimulating the internal carotid nerve. The postganglionic fibres innervating the nasal mucosa had a similar threshold to those of the nictitating membrane, which may indicate that there are small myelinated fibres innervating the mucosa.The preganglionic compound nerve action potential had four major components, S₁–S₄. S₁, S₂ and usually S₃ fibres were associated with membrane contraction; S₂, S₃ and sometimes S₁ fibres were associated with nasal vasoconstriction; and S₃, usually S₂ and occasionally S₁ fibres were associated with vasoconstriction in the tongue. It is concluded that each of these three groups of nerve fibres, but not S₄ fibres, may include fibres associated functionally with the three effectors.There was a considerable difference between the relative amplitude of the responses of the three effectors elicited by stimulation of the cervical sympathetic nerve at frequencies between 0.2 and 2 Hz. Vasoconstrictor responses were relatively larger than membrane contractions suggesting differences in the mechanisms of neurotransmission at the neuroeffector junctions.     

Investigating the construct validity of the SPAI-C: comparing the sensitivity and specificity of the SPAI-C and the SAS-A

Investigates the construct validity of the Social Phobia and Anxiety Inventory for Children (SPAI-C) by comparing its sensitivity and specificity with another self-report measure of social anxiety, the Social Anxiety Scale for Adolescents (SAS-A). Participants were 252 adolescents (124 males and 128 females) 13-17 years old. Adolescents completed the SPAI-C and the SAS-A and were interviewed using the Anxiety Disorders Interview Schedule for DSM-IV: Child Version (ADIS-IV:C). Parents were also interviewed and composite diagnoses were formed. Youth were classified as socially phobic or non-anxious based on these composite diagnoses. By comparing clinical cutoff scores with diagnostic group classification, the sensitivity and the specificity of the SPAI-C and SAS-A were compared. Results indicated that the SPAI-C was a more sensitive measure than the SAS-A (61.5% vs. 43.6%) providing evidence of the scale's construct validity. The two measures were similar with regard to specificity (82.7% for both). Implications of these results for assessment and research are discussed.     

Afferent and sympathetic innervation of the dome and the base of the urinary bladder of the female rat.

The afferent and sympathetic innervation of different regions of the urinary bladder (bladder dome vs. bladder base) was examined in the female rat using simultaneous injections of two fluorescent tracers. Retrogradely labeled cells were found in the dorsal root ganglia (DRG; L1-L3 and L6-S1), the sympathetic chain (SC; T12-L6), the inferior mesenteric ganglia (IMG) and the major pelvic ganglia (MPG). There were very few double-labeled cells, indicating that the dome and the base of the bladder receive innervation (afferent or sympathetic) from separate and distinct neuronal populations. Most of the sympathetic innervation of the bladder arose from the SC (dome: 77%; base: 89%) and it was carried equally by the hypogastric and pelvic nerves. The distributions of SC postganglionic neurons innervating the dome and the base of the bladder were very similar. In contrast, the contribution of IMG neurons was almost entirely restricted to the dome of the bladder (22%), with less than 1% innervating the base. Tyrosine hydroxylase-immunoreactive (TH) neurons in the MPG displayed a strong sexual dimorphism. Many TH neurons were found in the male MPG, but very few in the female MPG. In the female, these TH neurons projected almost exclusively to the bladder base of the female rat and were responsible for 10% of the sympathetic innervation of the base. Less than 1% innervated the dome. Therefore, prevertebral ganglia (IMG and MPG) show a strong regional selectivity in the innervation of the bladder of the female rat. The possible functional implications of this organization are discussed.