白花丹参丙酮酸脱羧酶基因的克隆和表达分析

家独行菜是世界多个地区的传统药物,从中分离的化学成分主要有生物碱、黄酮类、萜烯和芥子油苷等,现代药理学研究表明家独行菜具有降血糖、降血压、治疗胃肠病和骨折等活性。综述了家独行菜的化学成分和药理作用,为明确其药效成分以及作用物质基础提供参考和依据。     

岩黄连的抗乙肝病毒活性成分研究

目的 研究岩黄连Corydalis saxicola 生物碱中具有抗乙肝病毒活性的化学成分。方法 利用柱色谱法分离纯化岩黄连总生物碱中的化学成分,根据化合物的理化性质与波谱数据进行结构鉴定;利用HepG 2.2.15细胞株对质量分数较高的化学成分进行抗乙肝病毒活性试验。结果 从岩黄连总生物碱中分离鉴定了16个生物碱,分别为二氢血根碱（1）、d-紫堇碱（2）、旋卡文定碱（3）、stylopine（4）、6-丙酮基-5, 6-二氢血根碱（5）、二氢白屈菜红碱（6）、四氢巴马汀（7）、adlumidine（8）、(?)-salutaridine（9）、巴马汀（10）、原阿片碱（11）、小檗碱（12）、coptisine（13）、thalifaurine（14）、dehydroapocavidine（15）和木兰花碱（16）。对质量分数较高的化合物5、6、8～11、13、16进行抗乙肝病毒活性试验,结果表明化合物5、8活性一般,化合物11、16活性较弱,化合物6对HBV具有较强的抑制作用,化合物9次之。结论 化合物1、4～6、8、9、13、14和16均为首次从该药用植物分离得到,化合物10的质量分数较高,是该植物的主要成分,化合物6对HBV抑制活性较强,可能是该植物主要的活性成分。     

野西瓜总生物碱诱导 HepG2 细胞凋亡与［Ca2+］i 变化的相关性

目的 探讨野西瓜总生物碱 (Capparis spinosa alkaloid, CSA) 对人肝癌 HepG2 细胞的杀伤和诱导凋亡的作用机制。方法 MTT 法研究 CSA 对人肝癌 HepG2 的杀伤作用;荧光显微镜观察 HepG2 细胞形态;流式细胞仪研究 CSA 对 HepG2 细胞的凋亡诱导作用;激光共聚焦显微镜检测 CSA 对 HepG2 细胞内［Ca²⁺］_i 的影响。结果 CSA 对人肝癌 HepG2 有明显细胞毒性作用,并且有剂量依赖性,IC₅₀ 为 142.82 μg/mL;HepG2 细胞在 CSA 作用后出现特征性凋亡形态特征,凋亡细胞比率明显高于自然凋亡率;且阻断细胞由S期向 G₂ 期的移行,CSA 明显升高 HepG2 细胞［Ca²⁺］_i,并与药物质量浓度呈量效正相关。结论 CSA 对人肝癌 HepG2 有明显的杀伤和促凋亡作用,其机制可能与 CSA 造成肿瘤细胞内钙离子超载有关。     

基质辅助激光解吸电离成像质谱法可视化分析制川乌炮制过程生物碱空间分布的研究

目的 采用质谱成像显微镜可视化分析制川乌炮制过程中6个生物碱的变化情况。方法 准备不同炮制时间(9个时间点)的制川乌样品,用明胶溶液包埋后冷冻,制备30 μm厚度的切片样品进行实验。根据乌头碱、次乌头碱、新乌头碱、苯甲酰新乌头原碱、苯甲酰次乌头原碱和苯甲酰乌头原碱共6种标准品的质谱信号,选取α-氰基-4-羟基肉桂酸为基质,采用升华法涂覆基质后再进行喷雾。质谱数据采集质量范围为m/z 200~800,使用DHB溶液在m/z 100~900进行质量校正,以1 000 Hz频率的正离子模式进行基质辅助激光解吸-质谱成像(MALDI-MSI)。通过超高效液相色谱(UHPLC)测定不同炮制时间样品中6个生物碱的含量,用含量测定结果验证质谱成像结果的准确性。结果 质谱成像技术可以区分制川乌不同炮制程度的样品,并且可以实现生物碱空间分布的可视化分析。结论 质谱成像技术可运用于制川乌炮制过程质量研究,为中药炮制研究提供一种新思路。     

浙贝母实生鳞茎中总生物碱含量的测定

测定了中药浙贝母不同生长年限、不同品种及不同栽培地点实生鳞茎中总生物碱的含量,结果表明符合药典要求,可供药用,为有性繁殖应用于生产提供依据。     

Fagaronine,a novel antileukaemic alkaloid

A review is presented about fagaronine, a benzophenanthridine alkaloid responsible for antitumoural activity. Plant source, extraction, biosynthesis, medicinal properties, especially antileukaemic activity, are described.     

Antagonism of the aconitine-induced inexcitability by the structurally related Aconitum alkaloids, lappaconitine and ajacine

Aconitine, lappaconitine and ajacine are structurally related alkaloids occurring in several species of the Aconitum genus. While aconitine is known to activate the voltage-dependent sodium channel, lappaconitine has been reported to block this channel. To investigate a possible antagonism of the aconitine action on neuronal activity by lappaconitine and the closely related alkaloid ajacine, we have performed extracellular recordings of stimulus evoked population spikes and field excitatory postsynaptic potential (EPSP) in rat hippocampal slices. Aconitine (10–100 nM) diminished the amplitude of the orthodromic population spike in a concentration-dependent manner. When aconitine was applied in presence of 10 μM lappaconitine, the concentration–response curve was shifted to the right. Furthermore, the complete suppression of the population spike evoked by 100 nM aconitine was reversed by 10 μM lappaconitine. The action of lappaconitine was mimicked by ajacine, however, the latter alkaloid was less potent. Both lappaconitine and ajacine shifted the input–output relationship of the presynaptic fiber spike as function of the stimulation intensity and of the field EPSP as function of the presynaptic fiber spike to the right. After pharmacological isolation, the presynaptic fiber spike was decreased by both compounds in a frequency-dependent manner indicative for a use-dependent action. Thus, electrophysiologically these alkaloids seem to inhibit predominantly the excitability of the afferent fibres and, in consequence, neurotransmission between Schaffer collaterals and the CA1 neurons, thereby suppressing the firing of the latter. Ajacine and lappaconitine inhibited stimulus-triggered epileptiform population bursts in area CA1 elicited by omission of Mg²⁺ as well as spontaneously occurring epileptiform discharges in area CA3 elicited by omission of Mg²⁺ and elevation of K⁺. It is concluded that the inhibitory and antiepileptiform effect of ajacine and lappaconitine is mediated by a frequency-dependent inhibition of the voltage-dependent sodium channel, thereby decreasing the excitability which might be important for filtering high frequency bursts of action potentials characteristic for epileptiform activity in the hippocampus. Moreover, these alkaloids are naturally occurring antagonists of the sodium channel activator aconitine.     

Effect of Liposome—Encapsulated Total Alkaloid of Harmaline on Rabbit Lens Epithelial Cells：Experimental Study on the Prevention of Posterior Capsule Opacification

Purpose: To investigate whether liposome encapsulated total alkaloid of Harmaline (TAH) as a therapeutic agent is beneficial to prevention of posterior capsular opacifi-cation (PCO).Methods: Liposome-encapsulated TAH was prepared by modified freeze-thawing method. 0. 1ml of liposome-encapsulated TAH (0. 2mg/ml) was injected into the capsular bag during extracapsular lens extraction (ECLE) of each eye in total 10 rabbit eyes. Blank liposome or balance salt solution (BSS) was used as control. Slit-lamp examination and histopathological examination was used to evaluated capsule opacifica-tion. Intraocular pressure (IOP) , density and morphology of corneal endothelia cells, the amplitude and latency of b wave of ERG were measured.Results: The inflammatory response was mild both in TAH treated and the control group. PCO formation occurred in the control group 2 weeks postoperatively, but the posterior capsule was clear in TAH treated eyes. 4 weeks and 8 weeks after operation, PCO occurred both in TAH treated     

Vinflunine (20′,20′-difluoro- 3′,4′-dihydrovinorelbine), a novel Vinca alkaloid, which participates in P-glycoprotein (Pgp)-mediated multidrug resistance in vivo and in vitro

Vinflunine (VFL) is a novel derivative of vinorelbine (NVB, Navelbine®), which has shown markedly superior antitumor activity to NVB, in various experimental animal models. To establish whether this new Vinca alkaloid participates in P-glycoprotein (Pgp)-mediated multidrug resistance (MDR), VFL-resistant murine P388 cells (P388/VFL) were established in vivo and used in conjunction with the well established MDR P388/ADR subline, to define the in vivo resistance profile for VFL. P388/VFL cells proved cross-resistant to drugs implicated in MDR (other Vinca alkaloids, doxorubicin, etoposide), but not to campothecin or cisplatin and showed an increased expression of Pgp, without any detectable alterations in topoisomerase II or in glutathione metabolism. The P388/ADR cells proved cross-resistant to VFL both in vivo and in vitro, and this VFL resistance was efficiently modulated by verapamil in vitro. Cellular transport experiments with tritiated-VFL revealed differential uptake by P388 sensitive and P388/ADR resistant cells, comparable with data obtained using tritiated-NVB. In various in vitro models of human MDR tumor cells, whilst full sensitivity was retained in cells expressing alternative non-Pgp-mediated MDR mechanisms, cross resistance was identified in Pgp-overexpressing cells. Differences were, however, noted in terms of the drug resistance profiles relative to the other Vincas, with tumor cell lines proving generally least cross-resistant to VFL. Overall, these results suggest that VFL, like other Vinca alkaloids, participates in Pgp-mediated MDR, with tumor cells selected for resistance to VFL overexpressing Pgp, yet MDR tumor cell lines proved generally less cross resistant to VFL relative to the other Vinca alkaloids.     

长春花叶片愈伤组织诱导及培养过程中生物碱合成类型的变化和调节及有关酶的分析

在４种不同培养基上诱导长春花叶片愈伤组织发生的过程中,叶片中含量较高的文多灵（ｖｉｎｄｏｌｉｎｅ）和长春质碱（ｃａｔｈａｒａｎｔｈｉｎｅ）迅速降解至极低水平,而在叶片中含量较低的阿玛碱（ａｊ－ｍａｌｉｃｉｎｅ）和蛇根碱（ｓｅｒｐｅｎｔｉｎｅ）的合成却逐渐增加,同时酸性和碱性过氧化物酶（ｐｅｒｏｘｉｄａｓｅ）活性呈上升趋势,尤其碱性过氧化物酶与蛇根碱和阿玛碱的相关性较密切。光照和黑暗中的变化趋势基本一致,但光照可提高过氧化物酶的活性,对培养物的长春质碱、文多灵和蛇根碱的合成有利。不同的培养基对生物碱的合成影响较大,２,４－Ｄ显著抑制生物碱的合成,持续光照使愈伤组织的生长受到抑制,使阿玛碱向其蛇根碱转化。