清风丸对偏头痛患者血小板5-HT2A受体和粘附分子表达影响的临床研究

目的:观察清风丸对偏头痛血小板5-羟色胺2A(5-HT2A)受体mRNA和粘附分子表达的影响方法:将68例偏头痛患者随机分为2组。治疗组35例,口服清风丸(由川芎、当归、白芍、红花、羌活、全蝎、防风、附子、细辛、麻黄、钩藤、黄芪、泽泻、生龙骨等组成)治疗;对照组33例,口服氟桂嗪胶囊治疗。2组均以30天为1疗程。主要观察临床疗效、血小板5-HT2A受体和粘附分子的表达。结果:显效率、总有效率治疗组分别为37.14%、94.29%,对照组分别为15.15%、60.61%。2组比较,差异有显著性或非常显著性意义(P<0.05,P<0.01)。血小板5-HT2A受体mRNA表达,对照组发作期显著高于对照组间歇期、治疗组和健康组(P<0.01);治疗组与对照组(发作期、间歇期)比较,差异均有非常显著性意义(P<0.01) 对照组发作期5-HT2A受体mRNA表达水平与血小板膜粘附分子CD41呈高度正相关,与CD62P呈中度正相关。结论:清风丸可能通过降低血小板5-HT2A受体和粘附分子的表达抑制血小板活化,从而达到治疗的目的。     

新正天丸对偏头痛患者血小板5-HT2A受体和粘附分子表达的影响

目的探讨新正天丸对偏头痛患者血小板5-HT2A受体mRNA和粘附分子表达的影响及其相互的关系。方法将68例患者分为2组,治疗组(35例)口服新正天丸,对照组(33例)口服氟桂嗪胶囊。利用RT-PCR技术检测偏头痛患者血小板中的5-HT2A受体mRNA,应用流式细胞仪检测偏头痛患者全血血小板膜CD41和CD62P。结果治疗组疗效优于对照组。偏头痛患者5-HT2A受体mRNA的表达发作期与正常人有显著差异,经新正天丸治疗后可显著降低其表达。偏头痛患者发作期、间歇期血小板膜粘附分子表达均较正常人为高,而新正天丸能明显降低其表达。结论偏头痛患者5-HT2A受体mRNA的表达和血小板膜粘附分子的表达水平均显著增强,新正天丸可能通过降低上述指标的表达抑制血小板活化,从而达到治疗目的。     

加味四逆散对慢性应激大鼠海马区5羟色胺2A受体(5-HT2AR)及其mRNA表达的影响

目的:探讨加味四逆散对慢性应激模型大鼠海马区5-HT2A受体及其mRNA表达的影响。方法:选择Open-feild法评分相近的74只成年SD大鼠,随机分为对照组、模型组、氟西汀组、加味四逆散高,中,低剂量组。应用分养和长期不可预见的中等强度应激造成大鼠抑郁模型,以免疫组织化学法和原位杂交法研究大鼠海马区5-HT2AR及其mRNA表达的变化。结果:与对照组比,模型组和加味四逆散高剂量组海马区5-HT2AR表达降低(P0.01,P0.05);模型组、氟西汀组和加味四逆散中剂量组5-HT2AR mRNA表达降低(P0.01,P0.05)。与模型组比,各治疗组5-HT2AR及mRNA的表达均升高(P0.05,P0.01)。各组之间5-HT2AR/mRNA的变化无统计学差异。结论:慢性应激能诱发大鼠轻度抑郁障碍和海马区5-HT2AR及其mRNA的低表达。加味四逆散改善慢性应激大鼠的抑郁症状可能与调节海马区5-HT2A受体系统增高其表达有关。     

四逆散冻干粉对睡眠剥夺果蝇头部5-HT含量和5-HT_(1A)受体表达的影响

目的:探讨四逆散冻干粉改善果蝇睡眠作用的机制。方法:收集12 h内羽化未交配的野生型Canton S品系黑腹果蝇,小气流CO2麻醉,雌雄分开,雌雄分别随机分为空白对照组、睡眠剥夺模型组和给药组,每组30只果蝇,利用ELISA法分别测定各组果蝇头部的5-HT的含量。利用实时定量PCR法,测定各组果蝇头部5-HT1A受体的表达差异。结果:四逆散冻干粉分别能提高雌性与雄性果蝇头部5-HT的含量,与模型组相比具有极显著性差异(P0.01)。四逆散冻干粉还能增加果蝇头部5-HT1A受体的表达量。结论:四逆散冻干粉改善果蝇睡眠的作用机制与增加果蝇头部5-HT含量和促进果蝇头部5-HT1A受体表达有关。     

慢性应激对大鼠海马区5-HT2AR及其mRNA表达的影响

目的：探讨慢性应激对大鼠海马区5-HT2A受体及其mRNA表达的影响。方法：选择open-feild法评分相近的20只成年SD大鼠随机分为对照组和模型组。应用分养和长期不可预见的中等强度应激造成大鼠抑郁模型，以免疫组化法和原位杂交法检测大鼠海马5-HT2AR及其mRNA的表达。结果：（1）模型组体重增长14d时明显低于对照组。（2）模型组水平活动和垂直活动于第7天后明显降低；糖水消耗量于第14天后明显减少。（3）模型组海马5-HT2AR及其mRNA表达均低于对照组（均P〈0．01）。结论：慢性应激能诱发大鼠轻度抑郁，其机制与海马5-HT2AR及其mRNA的低表达有关。     

针刺对5-HT致痒大鼠IgE与5-HT2A受体的影响

[目的]观察针刺对5-HT致痒大鼠的止痒效果,探讨针刺对5-HT致痒大鼠的行为学指标,以及血清中IgE与皮肤中5-HT2A受体的影响。[方法]64只大鼠随机分为8组,分别为空白对照组、模型组、阳性药物组、常规治疗组、针刺治疗1组、针刺治疗2组、中药治疗1组、中药治疗2组,每组8只。空白对照组和模型对照组予生理盐水灌胃,阳性药物组予喂饲酮舍林,常规治疗组予喂饲开瑞坦,针刺治疗1组针刺委中等穴位,针刺治疗2组针刺血海等穴位,中药治疗1组予当归、生地等中药灌胃,中药治疗2组予乌梅等中药灌胃。取空白对照组大鼠于其颈背部皮内注射10μL0.9%生理盐水,取其余各组大鼠于其颈背部皮内（肩胛连线中点上方5mm处）注射10μL2%5-HT（94mmol/L）溶液。建立瘙痒模型后,观察大鼠搔抓次数,并测定大鼠血清中IgE和皮肤中5-HT2A受体含量。[结果]模型组大鼠血清IgE升高,皮肤内5-HT2A受体升高,搔抓次数增加;两组针刺治疗组大鼠血清IgE降低,皮肤内5-HT2A受体降低,搔抓次数减少。[结论]针刺治疗可降低5-HT致痒大鼠血清中IgE和皮肤内5-HT2A受体的浓度,对其搔抓症状有明显的抑制作用。     

针刺对5-HT致痒大鼠IgE与5-HT2A受体的影响

[目的]观察针刺对5-HT致痒大鼠的止痒效果,探讨针刺对5-HT致痒大鼠的行为学指标,以及血清中IgE与皮肤中5-HT2A受体的影响。[方法]64只大鼠随机分为8组,分别为空白对照组、模型组、阳性药物组、常规治疗组、针刺治疗1组、针刺治疗2组、中药治疗1组、中药治疗2组,每组8只。空白对照组和模型对照组予生理盐水灌胃,阳性药物组予喂饲酮舍林,常规治疗组予喂饲开瑞坦,针刺治疗1组针刺委中等穴位,针刺治疗2组针刺血海等穴位,中药治疗1组予当归、生地等中药灌胃,中药治疗2组予乌梅等中药灌胃。取空白对照组大鼠于其颈背部皮内注射10μL0.9%生理盐水,取其余各组大鼠于其颈背部皮内(肩胛连线中点上方5mm处)注射10μL2%5-HT(94mmol/L)溶液。建立瘙痒模型后,观察大鼠搔抓次数,并测定大鼠血清中IgE和皮肤中5-HT2A受体含量。[结果]模型组大鼠血清IgE升高,皮肤内5-HT2A受体升高,搔抓次数增加;两组针刺治疗组大鼠血清IgE降低,皮肤内5-HT2A受体降低,搔抓次数减少。[结论]针刺治疗可降低5-HT致痒大鼠血清中IgE和皮肤内5-HT2A受体的浓度,对其搔抓症状有明显的抑制作用。     

大鼠模拟经前期综合征肝气逆证不同脑区5-HT1A受体基因表达mRNA水平变化

本文以大鼠模拟经前期综合征（PMS）肝失疏泄所导致的始发证候肝气逆证为切入点,运用现代分子生物学技术,对肝失疏泄产生肝气逆证时的主要改变指标五羟色胺（5-HT1A）受体基因表达在中枢不同脑区（下丘脑、边缘叶）mRNA水平上的变化进行了探讨。结果显示,在肝失疏泄产生肝气逆证时中枢神经系统中5-HT1A受体的基因表达在mRNA水平均受到一定的抑制,即在肝失疏泄产生肝气逆证时5-HT1A受体的活性明显下降,且下丘脑中5-HT1A受体的活性要比边缘叶中5—HT1A受体的活性下降得显著。     

5-HT1A受体对癫痫伴发抑郁的影响及神经保护研究

目的 通过建立氯化锂-匹罗卡品癫痫伴发抑郁的模型试验,评价5-HT1A受体激动剂8-OH-DPAT治疗癫痫伴发抑郁的疗效.方法 建立癫痫伴发抑郁大鼠模型,随机分为5组,生理盐水组、模型组、卡马西平组及卡马西加低、高剂量8-OH-DPAT组,分别给予相应药物,通过尼氏染色及胶质纤维酸性蛋白免疫组化的方法观察抗癫痫的疗效,通过强迫游泳,开放视野的行为学方法、荧光定量PCR方法测定5-HT1A受体表达量来评定抗抑郁的效果.结果 5-HT1A受体激动剂与卡马西平联合使用,且能明显的改善抑郁行为、可以增高5-HT1A受体mRNA的含量,同样也具有对抗神经元凋亡和胶质细胞活化的作用（P〈0.05）.结论 5-HT1A受体激动剂不仅具有抗癫痫和抑郁的作用,而且也具有神经保护的作用.     

开心解郁汤对抑郁症模型大鼠脑皮质5-HT水平和海马与大脑皮层5-HTA亚型受体 mRNA表达的影响

目的:观察具有培元开郁作用的“开心解郁汤”对大鼠脑皮质中5-羟色胺(5-HT)水平和海马及大脑皮层5-HT1A受体和5 -HT2A受体mRNA表达以及行为学能力的影响.方法:SD大鼠随机分为正常对照组、抑郁症组、抑郁症+中药组、抑郁症+西药组,采用慢性不可预见性温和应激方法(CUMS)造成抑郁症大鼠模型.造模同时开始给药,中药组给予开心解郁汤(按生药量计,22 g·kg-1,西药组给予盐酸氟西汀2.4 mg·kg-1,其余各组给同体积生理盐水,每日1次,连续ig给药22 d.分别于实验前、抑郁1,8,15,22 d记录大鼠体重;并观察大鼠行为学变化.于22 d后分取脑皮质,应用高效液相色谱技术分析脑皮质中5 -HT水平变化;于22 d后分取脑皮质和海马区,采用实时荧光定量PCR技术,测定大鼠海马及脑皮质中5-HT1A和5-HT2A受体mRNA水平改变.结果:与正常对照组相比,抑郁造模各组体重增长明显减慢,体重增加数和糖水消耗量均下降,旷场实验测定水平活动和垂直活动次数均下降(P＜0.01,P＜0.05),给与中药干预后,大鼠体重增长有所增快,与抑郁模型组比较有显著差异.糖水消耗量、水平活动和垂直活动次数均较模型组明显增加(P＜0.01,P＜0.05);同时抑郁造模后5-HT总体水平呈现下降,给予中药和西药治疗后5-HT水平有显著升高(P＜0.05).模型组大鼠海马组织中5-HT1AR mRNA水平下降,而5-HT2AR mRNA水平升高,中药及西药治疗后均可不同程度地降低;而在脑皮质中5-HT1AR和5-HT2AR mRNA水平的改变则呈现相反的变化.结论:CUMS可引起大鼠行为学改变并致体重减轻和行为学改变,造成大鼠抑郁模型.开心解郁汤可以降低海马中5-HT1A受体mRNA和脑皮质中5-HT2A受体mRNA水平,同时升高海马中5-HT2A受体mRNA和脑皮质中5-HT1A受体mRNA水平,而部分实现其对抑郁症的治疗作用.     

补肾调肝清心方对围绝经期抑郁症睡眠障碍大鼠模型海马5-羟色胺1A受体、5-羟色胺2A受体的影响

目的：观察补肾调肝清心方对围绝经期抑郁症睡眠障碍模型大鼠海马5-羟色胺1A受体（5-hydroxytryptamine 1A receptor,5-HT1AR）、5-羟色胺2A 受体（5-hydroxytryptamine 2A recep-tor,5-HT2AR）的影响。方法将60只围绝经期大鼠按体重随机分为5组,每组12只：围绝经期正常对照组（ A）;围绝经期抑郁症睡眠障碍模型组（ B）;围绝经期抑郁症睡眠障碍补佳乐治疗组（ C）;围绝经期抑郁症睡眠障碍米氮平治疗组（ D）;围绝经期抑郁症睡眠障碍中药治疗组（ E）;A组为围绝经期正常大鼠,其余4组进行18天不可预知慢性应激。于应激后对造模动物进行72小时连续快速眼相睡眠剥夺（ REM）。每天在应激前1小时按组给药。在末次灌胃24小时后,颈椎脱臼法处死大鼠,取大鼠海马,分别用荧光定量检测海马5-HT1ARmRNA和5-HT2ARmRNA、蛋白印迹和免疫荧光检测海马5-HT1AR和5-HT2AR的表达。结果蛋白印迹检测（ western blot）海马及免疫荧光检测海马CA1区均显示：海马区5-HT1AR表达水平各组无明显差异（P〉0．05）,而5-HT2AR表达水平上,模型组显著低于正常组（P〈0．05）,治疗组提高了5-HT2AR的表达水平,且与模型组相比存在显著性差异（P〈0．05）;RT-PCR显示：海马区5-HT1AR mRNA水平各组无明显差异（P〉0．05）,而5-HT2AR mRNA水平,模型组明显低于正常组（P〈0．01）,米氮平组较模型组有极显著性差异（P〈0．001）,中药组较模型组有显著性差异（P〈0．05）。结论补肾调肝清心方能提高5-HT2AR的mRNA及蛋白水平的表达,免疫荧光的结果表明在海马CA1区发挥作用,表明补肾调肝清心方可以通过调节海马CA1区5-HT2AR表达来改善睡眠障碍。     

电针对慢性应激抑郁模型大鼠大脑皮层5HT1和5HT2受体数量和结合活性的影响

目的探讨电针"百会"足三里"穴对抑郁症的调节作用.方法复制慢性应激抑郁大鼠模型,施以电针"百会"足三里"穴治疗.采用美国GraphPad Prism 3.02专用统计软件,以非线性回归方法计算最大结合率(Bmax)和平衡解离常数(Kd),推测5-HT1A和5-HT2A的受体数量与结合活性.结果(1)慢性应激抑郁模型大鼠大脑皮层5-HT1受体的Bmax值明显低于正常组(P<0.01),电针可增加其Bmax值.(2)抑郁模型大鼠大脑皮层5-HT2受体的Bmax值明显高于正常组(P<0.01),电针可降低其Bmax值;抑郁模型大鼠5-HT2受体的Kd也高于正常组(P<0.05),电针治疗后Kd值也有所下降.结论5-HT受体功能不平衡是导致抑郁症的重要原因之一,电针"百会"足三里"穴有明显的调节作用.     

New antiinfective and human 5-HT2 receptor binding natural and semisynthetic compounds from the Jamaican sponge Smenospongia aurea

In addition to the sesquiterpene-phenol aureols (1), 6'-chloroaureol (2), and aureol acetate (3), eight indole alkaloids including the new N-3'-ethylaplysinopsin (9) have been isolated from the Jamaican sponge Smenospongia aurea. Makaluvamine O (10), a new member of the pyrroloiminoquinone class, was also isolated. The structures were characterized by spectroscopic methods, and two new derivatives of aureol were prepared to optimize the biological activity. Aureol N,N-dimethyl thiocarbamate (1a) and 6-bromoaplysinopsin (7) exhibit significant antimalarial and antimycobacterial activity in vitro. Compound 6 showed activity against the Plasmodium enzyme plasmepsin II. The 6-bromo-2'-de-N-methylaplysinopsin (6), 6-bromoaplysinopsin (7), and N-3'-ethylaplysinopsin (9) displaced high-affinity [(3)H]antagonist ligands from cloned human serotonin 5-HT(2) receptor subtypes, whereas the other compounds tested did not. Remarkably, the 6-bromo-2'-de-N-methylaplysinopsin (6) showed a > 40-fold selectivity for the 5-HT(2C) subtype over the 5-HT(2A) subtype.     

5-Hydroxytryptamine2A receptor binding activity of compounds from Litsea sessilis

A 96-well microplate filtration based 5-HT(2A) receptor-radioligand binding assay was optimized and adopted to carry out a bioassay-guided fractionation of the methanol extract of the leaves of Litsea sessilis. This purification led to the isolation of two compounds identified as (+)-boldine (1) and (+)-dehydrovomifoliol (2). (+)-Boldine binds to 5-HT(2A) receptors at high concentrations with a K(i) value of 2.16 microm. However, (+)-dehydrovomifoliol showed minimal competitive inhibition on the binding of [(3)H]ketanserin to the same receptor with a K(i) value of 2.06 mm. These results suggest that (+)-boldine influences the activity of 5-HT(2A) receptors through competitive binding as an agonist or antagonist.     

心理疏导疗法对脑卒中后抑郁症患者临床疗效及血浆NE、5- HT、DA的干预效应

目的:观察心理疏导疗法对脑卒中后抑郁症患者的临床疗效及血浆NE、DA、5- HT的干预效应.方法:采用解释、鼓励、支持安慰等方法来疏导患者的心理,治疗2周和4周后对比临床疗效积分的变化及比较治疗前、治疗2周、4周后静脉血血浆NE、DA、5-HT的含量变化.结果:治疗4周后的临床疗效明显优于治疗2周(P＜0.05);血浆NE含量治疗2周后较治疗前明显改善(P＜0.05),但和治疗4周后差别无统计学意义;DA、5- HT治疗2周后和治疗前的含量差别无统计学意义,但治疗4周后的含量较2周后明显改善(P＜0.05).结论:心理疏导疗法能改善脑卒中后抑郁症患者的临床疗效和血浆NE、DA、5-HT的变化.     

Involvement of 5-HT1A in the anxiolytic-like effect of dichloromethane fraction of Pimenta pseudocaryophyllus

Ethnopharmacological relevance

Medicinal applications of Pimenta pseudocaryophyllus infusion as a diuretic and aphrodisiac agent as well as tranquilizer in the form of tea for the treatment of emotional tension in Brazilian folk medicine has been in practice since time immemorial. Despite its popular therapeutic acceptance and claims, there are scanty scientific reports to corroborate its central biological activities.

Aim

To characterize anxiolytic-like effect of the dichloromethane fraction (DF) obtained from ethanolic leaf extract of the Pimenta pseudocaryophyllus and identify mechanisms of action involved while seeking to support its popular use as a soothing agent.

Material and methods

Mice (25–35 g) were treated orally with DF obtained from ethanolic leaf extract of Pimenta pseudocaryophyllus and were submitted to light-dark box (LDB) and elevated plus maze (EPM) tests. Different groups of mice were treated with flumazenil and NAN-190 to identify mechanisms of action involved in the anxiolytic-like effect of DF.

Results

Treatment with DF increased number of transitions and time spent in the light compartment of the LDB while the time spent and numbers of entries in the open arm of the LCE were significantly increased. Pre-treatment of the animal with flumazenil (2 mg/kg, i.p. – competitive antagonist of benzodiazepine site of GABA_A receptor) did not block this effect, thereby excluding participation of benzodiazepine site of the GABA_A receptor. However, anxiolytic-like effect of DF was reversed by pre-treatment with NAN-190 (0.5 mg/kg, i.p. – an antagonist of the 5-HT_1A receptor) thereby suggesting involvement of 5-HT_1A receptor. The thin layer chromatography and high-performance liquid chromatography analysis indicated the predominance of (E)-methyl isoeugenol and oleanolic acid in DF.

Conclusion

These results support the popular use of Pimenta pseudocaryophyllus as a calming agent and suggest the involvement of 5-HT_1A receptor.