天麻多糖的降血压作用

目的从天麻中提取出天麻多糖(PGB),研究PGB对高血压模型大鼠的降血压作用。方法水提醇沉法提取出天麻粗多糖,DEAE-52纤维素柱分离纯化得PGB。用两肾一夹(2K1C)方法制作高血压模型(RHR)大鼠,以卡托普利(10mg/kg·d)为阳性对照组,以服用自来水为阴性对照组,以天麻多糖低、中、高3个浓度(50、100、200mg/kg·d)给药30d后测定大鼠心率与血压的变化,尿量的变化,以及血清中一氧化氮(NO)、血浆内皮素(ET)和血管紧张素Ⅱ(AngⅡ)水平的变化。结果天麻多糖PGB能明显降低RHR大鼠的收缩压和舒张压(P<0·05),尤其是中、高剂量组(P<0·01),并且在剂量范围内呈剂量依赖性;对大鼠心率及尿量没有明显作用;血清NO含量(51·9±9·8vs对照组40·7±10·2)μmol/L,血浆ET含量下降(164±25vs对照组209±10)μmol/L,AngⅡ含量下降(247±20vs对照组310±37)μmol/L,P值均<0·01。结论天麻多糖对RHR大鼠具有良好的降压作用,其作用机制与促进内源性舒血管物质的生成及抑制内源性缩血管物质的释放,最终恢复二者拮抗效应的平衡有关。     

天麻通脑颗粒对急性脑梗死大鼠血管神经再生的影响及作用机制

目的 探讨天麻通脑颗粒对大脑中动脉梗死大鼠血管神经再生的作用机制.方法 取成年雄性SD大鼠54只,随机分为假手术组(n=18)和模型组(n=36).模型组大鼠采用线拴法制作大鼠大脑中动脉局灶性缺血再灌注模型,术后随机分为模型对照组(n=18)和给药组(n=18),于24 h后分别通过腹腔注射给予等量等渗生理盐水或天麻通脑颗粒,4ml/次,2次/d,共14d.术后2h、1d、7d、14d四个不同时间点对脑梗死后大鼠神经功能恢复程度进行神经功能缺损评分.采用苏木素-伊红染色法对脑梗死大鼠于术后第14天进行梗死灶体积测定.采用免疫组化法于术后第14天对缺血侧室管膜下区(SVZ)及纹状体进行5-溴脱氧尿核苷(BrdU)阳性细胞计数.结果 与模型组相比,给药组从术后第7天开始大鼠神经功能就有显著恢复(P=0.043,P=0.031).模型对照组与给药组大鼠的梗死灶体积差异有统计学意义(P=0.044).BrdU免疫组化显示,术后第14天给药组在SVZ及纹状体区域的BrdU阳性细胞数量较模型对照组有显著增高(P=0.009,P =0.044).结论 天麻通脑颗粒可以促进脑缺血大鼠神经功能的恢复,对缺血后脑细胞的增殖具有一定疗效.     

Training-induced changes in the expression of GABAA-associated genes in the amygdala after the acquisition and extinction of Pavlovian fear

Previous work suggests the gamma-aminobutyric acid (GABA)ergic system may be dynamically regulated during emotional learning. In the current study we examined training-induced changes in the expression of GABA(A)-related genes and the binding of GABA receptor radioligands in the amygdala after the acquisition and extinction of Pavlovian fear. Using in situ hybridization, we examined the expression pattern changes of mRNAs for GABAergic markers in the lateral, basolateral and central subdivisions of the amygdala in C57Bl/6J mice. These markers included GABA-synthesizing enzymes (GAD67 and GAD65), major GABA(A) receptor subunits (alpha1, alpha2, alpha3, alpha5, beta2 and gamma2) and the expression of mRNAs that are involved in a variety of GABA-related intracellular processes, including GABA transporter-1 (GAT1), GABA(A) receptor-associated protein and the GABA(A) clustering protein, gephyrin. With fear conditioning, we found decreased mRNA levels of alpha1, alpha5 and GAD67, as well as deceased benzodiazepine binding in the amygdala. Fear extinction induced an increase in mRNA levels of alpha2, beta2, GAD67 and gephyrin, as well as a decrease in GAT1. Together, these findings indicate that the acquisition of fear induced a downregulation of mRNA markers related to a decrease in amygdala GABAergic function, whereas the acquisition of fear extinction produced an upregulation of GABAergic markers related to enhanced GABAergic transmission.     

Method development and validation for the high-performance liquid chromatography assay of gastrodin in water extracts from different sources of Gastrodia elata Blume

Gastrodia elata Blume is commonly used as a medical herb in China for ameliorating headaches, dizziness, and convulsions. In previous studies, water extracts of G. elata Bl. (WGE) have demonstrated potential to act as therapeutic agents to improve depression-like symptoms in rats. As gastrodin (GAS) is a major active compound in WGE, its quantitation in WGE is important for quality control. The objective of this study was to develop an optimized and validated reversed-phase high-performance liquid chromatography method for the analysis of GAS in different sources of WGE. We evaluated the GAS content in varieties of G. elata Bl. including G. elata Bl. f. glauca S. Chow and G. elata Bl. f. elata. We also evaluated the GAS content of the latter variety from two different origins, Yun-nan and Hu-nan. The results indicate that the amount of GAS analyzed in WGE from G. elata Bl. f. glauca S. Chow is five times higher than that of G. elata Bl. f. elata from Yun-nan and Hu-nan. A significant difference in GAS content was observed between varieties of G. elata Bl., although not between locations of origin.     

Protective effects of several components of Gastrodia elata on lipid peroxidation in gerbil brain homogenates

Gastrodia elata (GE) Blume, a traditional herbal agent, has been used mainly in anticonvulsive treatment in Asia. Recently, extracts of GE were evaluated for their potential as neuroprotectives and antioxidants. This study was designed to examine the antioxidant effect of the ether fraction of the methanol extract (EFME) of GE along with its major constituents vanillin, vanillyl alcohol, hydroxybenzaldehyde and hydroxybenzyl alcohol. In experiment 1, gerbils were treated with EFME of GE at a dosage of 500 mg/kg/day for 2 weeks. Oxidative stress was induced with H(2)O(2) or ferrous ion, and lipid peroxidation was measured. In experiment 2, oxidative stress was induced with various concentrations of H(2)O(2) or ferrous ammonium sulfate, and lipid peroxidation was measured. To compare the antioxidant potency, the inhibitory concentration 50% (IC(50)) was determined. EFME of GE reduced auto-peroxidation and H(2)O(2)-induced lipid peroxidation. However, it did not significantly reduce ferrous ammonium sulfate-induced lipid peroxidation. The order of antioxidation potency was as follows: hydroxybenzyl alcohol > vanillyl alcohol > vanillin > hydroxybenzaldehyde. In the case of hydroxybenzaldehyde, its antioxidant effect was more potent than that of melatonin. The excellent antioxidant effects of GE and its main constituents may have potential in the treatment of lipid peroxidation-associated neurological disease.     

Neuropilin-1在胰腺导管癌及MIA PaCa-Ⅱ细胞系中的表达及意义

目的探讨Neuropilin-1在胰腺导管癌组织及MIAPaCa-Ⅱ胰腺癌细胞系中的袁达及意义。方法运用免疫组化和RT—PCR法分别检测正常胰腺组织、癌旁组织、胰腺癌组织及MIAPaCa-Ⅱ细胞系中Neuropilin-1蛋白及mRNA表达水平。结果蛋白水平可见正常胰腺组织无表达,癌旁组织轻度表达,而胰腺癌组织及MIAPaCa-Ⅱ胰腺癌细胞中高水平表达。神经组织也可表达Neuropilin-1。mRNA水平见正常胰腺组织微量表达,癌旁组织中度表达,而胰腺癌组织及MIAPaCa-Ⅱ胰腺癌细胞中高水平表达。结论Neuropilin-1可能参与了胰腺癌的发生发展,在胰腺癌神经转移中可能起着重要作用。     

脑源性神经营养因子预处理对大鼠脑缺血再灌注损伤氧化应激及神经细胞凋亡的影响

目的探讨不同剂量的脑源性神经营养因子（BDNF）预处理对大鼠局灶性脑缺血再灌注（I／R）损伤氧化应激及神经细胞凋亡的影响。方法采用大鼠大脑中动脉线栓法（MCAO）建立局灶性脑I／R损伤模型,BDNF于缺血前12h经侧脑室注射给药。采用黄嘌呤氧化酶法和硫代巴比妥酸法检测脑组织超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量,TUNEL法检测脑皮层凋亡神经细胞,免疫组化法检测Bcl-2、Bax蛋白表达。结果与I／R组比较,BDNF预处理各组脑组织SOD活性明显增强,MDA含量明显降低,给药各组中脑皮层Bcl-2蛋白表达明显增高,而Bax蛋白表达及TUNEL阳性细胞指数均明显降低（P〈0．01）;在BD-NF预处理各组中,以BDNF预处理高剂量组脑组织SOD活性及Bcl-2蛋白表达最高、MDA含量和Bax蛋白的表达以及TUNEL阳性细胞指数最低（P〈0．01）。结论不同剂量的BDNF预处理均能明显减轻脑I／R后氧化应激和神经细胞凋亡,具有不同程度的脑保护作用,其保护作用与BDNF预处理的剂量有关。其机制可能与BDNF预处理能够提高机体内源性抗氧化物质的含量及调节凋亡相关基因的不同表达有关。     

血见愁中1个新的倍半萜类化合物

目的 研究血见愁Teucrium viscidum地上干燥部分的化学成分,发现抗氧化活性化合物。方法 利用硅胶柱色谱、凝胶、ODS柱色谱等色谱技术方法进行分离纯化,并综合运用IR、HR-ESI-MS、¹H-NMR、¹³C-NMR、HSQC、HMBC、COSY等方法鉴定化合物的结构,利用超氧阴离子自由基清除实验评价其抗氧化活性;结果 从血见愁乙醇提取物醋酸乙酯萃取部分分离得到1个倍半萜类化合物(1)和6个苯丙素类化合物(2～7),分别鉴定为9-羟基-6,7-去环氧-茎点霉甜菜烯酮(1)、迷迭香酸(2)、迷迭香酸甲酯(3)、迷迭香酸乙酯(4)、反式咖啡酸(5)、反式咖啡酸乙酯(6)、反式咖啡酸乙烯酯(7),化合物2、3、5～7表现出一定的超氧阴离子清除活性,半数效应浓度(median effect concentration, EC50)分别为(2.07±1.22)、(0.95±0.26)、(0.84±0.18)、(1.98±1.12)和(5.39±0.89)μg/mL。结论 化合物1为新化合物,命名为血见愁酮A;化合物2～4为血见愁植物中首次分...     

Contribution of spinal 5-HT1A and 5-HT7 receptors to locomotor-like movement induced by 8-OH-DPAT in spinal cord-transected mice

Growing evidence from in vitro studies suggests that spinal serotonin (5-HT) receptor subtypes 5-HTR(1A) and 5-HTR(7) are associated with an induction of central pattern generator activity. However, the possibility of a specific role for these receptor subtypes in locomotor rhythmogenesis in vivo remains unclear. Here, we studied the effects of a single dose (1 mg/kg, i.p.) of 8-hydroxy-2-(di-N-propylamino)-tetralin (8-OH-DPAT), a potent and selective 5-HTR(1A/7) agonist, in mice spinal cord transected at the low-thoracic level (Th9/10). The results show that 8-OH-DPAT acutely induced, within 15 min, hindlimb movements that share some characteristics with normal locomotion. Paraplegic mice pretreated with the selective 5-HTR(1A) antagonists, WAY100,135 or WAY100,635, displayed significantly less 8-OH-DPAT-induced movement. A similar reduction of 8-OH-DPAT-induced movements was found in animals pretreated with SB269970, a selective 5-HTR(7) antagonist. Moreover, a near complete blockade of 8-OH-DPAT-induced movement was obtained in wild-type mice pretreated with 5-HTR(1A) and 5-HTR(7) antagonists, and in 5-HTR(7)-/- mice pretreated with 5-HTR(1A) antagonists. Overall, these results clearly demonstrate that 8-OH-DPAT potently induces locomotor-like movement in the previously paralysed hindlimbs of low-thoracic-transected mice. The results, with selective antagonists and knockout animals, provide compelling evidence of a specific contribution of both receptor subtypes to spinal locomotor rhythmogenesis in vivo.     

蒸制断生后真空冷冻干燥对天麻质量的影响

目的:提高真空冷冻干燥天麻的天麻素含量。方法:采用蒸制后再真空冷冻干燥的方法对天麻进行干燥,考察天麻的外观及微观结构,测定天麻复水率、多糖和天麻素含量及不同干燥工艺下天麻粉末在模拟胃肠液中的溶出量与溶出率。结果:与鲜天麻直接真空冷冻干燥相比,天麻经蒸制后再真空冷冻干燥其外观和切面微观结构无显著变化。95℃蒸制4 min后再真空冷冻干燥,其天麻素含量和复水率分别为0.875%和502%,显著高于鲜天麻直接冷冻干燥(0.280%和362%)和120℃蒸制4 min后再真空冷冻干燥(0.775%和466%),但3个干燥工艺下天麻的多糖含量无显著差异。热风干燥天麻及真空冷冻干燥天麻粉末的红外指纹图谱谱峰位置相近,峰形相似,说明天麻成分无显著变化。不同工艺干燥天麻的天麻素在模拟胃液和肠液中的溶出率无显著差异。结论:天麻经95℃蒸制4 min后再进行真空冷冻干燥可在保持其外观和微观形态的条件下显著提高天麻素含量和复水率,天麻真空冷冻干燥生产中可对该工艺进行适当修正以推广应用。