中药复方研究的思路和方法

本文阐述了近年来中药复方的药效物质基础（有效成分）,复方药理研究以及复方新药研制的思路和方法,对一些新观点与假设的实现途径进行了综述。     

基于“谱-效”相关的中药药效物质基础研究方法

中药作为我国传统医药学的重要组成部分,在我国传统的防治疾病中占有举足轻重的地位。中药主要来源于动植物、微生物及矿物,含有不同结构和性质的多种成分,其物质基础研究具有复杂性,目前尚处于初步探索研究阶段。近年来开展的中药谱效关系研究为其物质基础研究提供了思路和方法,重视将先进的科学技术与中药研究有机结合,进而为中药实现现代化提供了可能。本文通过查阅相关文献,总结了近年来有关中药谱效关系相关性研究思路和方法,对中药谱效关系研究方法进展作一综述,为其药效物质基础研究提供依据及借鉴。     

基于中药有效成分族信息辨识技术的中药物质基础

正随着中药现代化进程的不断深入,相关研究技术不断革新,中药所含化学成分的信息已不断清晰、完整.因此,现阶段,如何确定其中发挥药效、功效乃至药性的物质基础已经是中药现代化研究的关键科学问题之一.已有研究利用化学、生物学和药理学的方法和技术,从分子、细胞、器官和整体多个层次,获得了大量中药物质基础信息,为中药产品的研究     

用《组合中药学》的理论开发新型中药

我们以现代科技理论为出发点,提出了<组合中药学>新理论体系,试图使中外学者对中药的认识趋于一致.中药的本质就是"组合".无论是单方或复方都是一个庞大的分子库.具有相同或相似功能的分子组合成一个群体.这些群体表现出特定的药效."组合"有三层意思生药的组合、活性分子群的组合与特定疗效的组合.分子群是特定药效功能的物质基础.<组合中药学>就是从<组合分子>水平上研究中药及其药效的物质基础,阐明中药活性部位的相互作用与药效的相互关系及其作用机制.<组合中药学>的三个新概念中药功效分子族具有相似药效作用的一类分子的组合,其骨架及功能团相同或相似.单味中药表征性组合分子单味中药药效的代表性功效分子族,即主要药效分子组合.单味中药非表性组合分子单味中药表征性分子以外的其它功效分子族,即辅助药效分子的组合.<组合中药学>的主要研究范围中药药效的组合分子学基础(植物化学,天然产物);组合分子的药理学基础(分子生物学,细胞生物学,药理学).本文提出了应用<组合中药学>研究新型中药的新方法.     

《组合中药学》及其理论系统的建立

我们以传统中医药理论与现代科技的结合为出发点,提出了《组合中药学》新理论体系,试图解答中药现代化中基础理论方面的关键问题。中药的本质就是“组合”。无论是主或复方都是一个庞大的分子库,具有相同或相似功能的分子组合成一个群体。这些群体表现出特定的药效。“组合”有三层意思：生药的组合、分子群的组合与特定药效功能的组合。分子群是特定药效功能的物质基础。分子群的观念表明了东方医药不同于西方的本质之所在,西方医药学研究的对象是单一的化合物。复方是特定药效的组合,分别具有不同或相同或相似药效的几类分子群的组合。即为中药的复方制剂。这些分子群体的组合或相辅相成,或减毒增效,以达到治疗的目的。本论文阐述了《组合中药学》的目的、意义和研究方法。《组合中药学》作为一大研究领域,可以建立一整套现代中药学的新理论体系,为中药的现代化提供科学依据,并可以此作为中药现代化切入点,以解决中医药科学化的关键问题。遵循这一理论体系,可以使中医药研究系统化,起到提纲挈领的作用,具有重要的理论价值和指导意义。《组合中药学》就是从《组合分子》水平上研究中药及其药效的物质基础,阐明中药活性部位的相互作用与药效的相互关系及其作用机制。《组合中药学》的三个新概念：中药功效分子族：具有相似药效作用的一类分子的组合,其骨架及功能团相同或相似。中药表征性组合分子：单味中药药效的代表性功效分子族,即主要药效分子组合。中药非表征性组合分子：单味中药表征性分子以外的其他功效分子族,即辅助药效分子的组合。     

黄芩汤药效物质基础及其治疗胃肠道疾病研究进展

黄芩汤是《伤寒论》中治疗下痢的经典方剂,由黄芩、芍药、甘草(炙)和大枣四味中药煎煮而成,用于治疗痢疾、腹泻等胃肠道疾病已有近1800年,中药治疗胃肠道疾病相较于西药更安全。本文结合国内外相关文献,对近几年来黄芩汤药效物质基础研究进行概括总结,并对其治疗溃疡性结肠炎和减轻化疗药物胃肠道毒性机制的最新研究进行了综述,结合胃肠道疾病的发病机制以及黄芩汤减毒机制的研究方法对现有研究过程中存在的不足进行了评述,并对中药疗效研究展开设想,提出以组学作为研究手段,配合中药复方整体作用模式的组分-疗效研究思路,以期为黄芩汤治疗胃肠道疾病的研究提供参考。     

中国特有植物明党参化学成分和药理研究进展

本文报道中国特有名贵中药明党参ＣｈａｎｇｉｕｍｓｍｙｒｎｉｏｉｄｅｓＷｏｌｆ根的化学成分与药理研究的最新进展及应用历史概况,阐明了该药物治病物质基础,发现了新的活性成分和药效,提出了合理、科学、先进的质量检测指标和方法。为开发防治老年病的新药提供了实验基础。对明党参道地性研究亦具有重要意义。     

蛋白质组学在动物源中药研究中的应用

动物源中药是传统中药的重要组成部分，因其成分复杂不易分离分析，导致其活性成分和机制研究进度缓慢，也进一步增加了评价动物源药材的品种质量以及保障临床安全用药的难度。随着多维色谱、质谱、生物信息学等技术的飞速发展，蛋白质组学逐渐成为动物源中药研究的重要工具。蛋白质组学从整体水平对药材本身或药物处理前后的动物或细胞模型的总蛋白进行定性定量分析，在获得中药指纹图谱、挖掘潜在活性成分以及定位药物靶点和通路方面具有显著优势。本文主要介绍了蛋白质组学技术及其在动物源中药药材品种和质量鉴定、药用资源开发和可持续发展、药效物质基础及作用机制研究等方面的应用，为动物源中药的深入研究提供新的研究思路和方法。     

沃特世与黑龙江中医药大学共建中美中医方证代谢组学技术合作中心

正沃特世公司(Waters~)近日宣布与黑龙江中医药大学共同建立中美中医方证代谢组学技术合作中心。该中心将针对与药效物质基础相关的中药有效性及安全性等质量问题进行研究,探索基于临床有效性的中药创新药物发现。中医方证代谢组学是中医药现代化研究方法学意义上的创新,主要研究中药复方给药形式的特殊性及方证对应疗效的专属性。合作中     

基于中医农业理论创制新型生物农药的实践探索

中医农业是利用中医药基本理论和方法解决现代农业生产过程中遇到的诸多实际问题,是实现生态农业与有机农业的重要途径.本文分析了4个中医农业探索案例.首先论述了从“清热解毒”类传统中药复方及其单味药材中挖掘苦参碱、小檗碱和大黄素甲醚等植物源农药,提出了传统“清热解毒”类中药复方及其单味中药是新型植物源农药创制的重要来源;接着对中药天仙子中挖掘具有新颖结构的天仙子新碱以创制新型植物免疫诱抗剂和植物生长调节剂进行了阐述,从药效物质基础的角度对铁线莲属植物发挥“整体”药效以挖掘新颖结构农用活性分子的潜力进行了归纳;最后对猕猴桃溃疡病防治技术中的“治未病”策略、“两前两后”关键防控技术以及复方生物菌剂创制进行了阐述.以上中医农业实践探索为更好地利用中医药理论和资源创制新型生物农药提供了思路和借鉴.     

Influence of method of presentation on results of plant-host preference tests with two species of grasshopper

Leaf material of two Piper spp. (Piperaceae) was presented to two different generalist herbivores (Orthoptera: Acrididae) to determine whether method of leaf presentation in preference trials affects choice. For the grasshopper Leptomerinthoprora brevipennis, the method of presentation had no effect on preference ranking: more of Piper sancti-felicis was eaten than of P. arieianum, regardless of whether leaf material was presented as leaf disks, whole leaves supplemented with water, or whole leaves without water. In contrast, preference of the grasshopper Abracis flavolineata depended on presentation method. In that case, whole leaves of P. scancti-felicis supplemented with water were preferred over the same of P. arieianum, while dry weight consumption of leaf disks of P. arieianum was greater than that for disks of P. sancti-felicis. When presented with leaves not supplemented with water, A. flavolineata showed no significant preference for one plant species over another.

---

Résumé Des feuilles provenant de deux espèces de Piper (Piperacae) ont été proposées à deux Orthoptères, Acrididae, herbivores généralistes, pour déterminer si la méthode de présentation des feuilles modifiait le choix. Pour Leptomerinthoprora brevipennis, la méthode de présentation est sans influence sur l'ordre de préférence; la consommation de Piper sancti-felicis est toujours supérieure à celle de P. arieianum, quelque soit le mode de présentation: disques de feuilles, feuilles entières humidifiées, ou feuilles entières sans addition d'eau. A l'opposé, l'ordre de préférence de Abracis flavolineata dépend du type de présentation; les feuilles entières humidifiées de P. sancti-felicis sont préférées à celles de P. arieianum proposées de la même façon, tandis que la consommation de disques secs de P. arieianum est plus importante que celle des disques secs de P. sancti-felicis. En présence de feuilles entières humidifiées, A. flavolineata ne manifeste aucune préférence.

     

Diversity of the Carbohydrate Composition of the Antigenic Polysaccharides of Proteobacteria of the Genera Pseudoalteromonasand Marinomonas

The sugar analysis of the glycans of the type strains of marine proteobacteria of the genera Pseudoalteromonasand Marinomonas—Pseudoalteromonas atlanticaIAM12927^T, P. aurantiaNCIMB 2033^T, P. citreaATCC 29719^T, P. elyakoviiKMM 162^T, P. espejianaATCC 29659^T, P. piscicidaNCIMB 645^T, P. tetraodonisIAM 14160^T, Marinomonas communisATCC 27118^T, and M. vagaATCC 27119^T—showed that they contain glucose, galactose, galactosamine, glucosamine, fucose, rhamnose, mannose, heptose, 2-keto-3-deoxyoctonate (KDO), uronic acids, colitose (3,6-dideoxy-L-xylo-hexose), and 6-deoxy-L-talose. The carbohydrate composition of the antigenic polysaccharides (PSs) of P. elyakoviiKMM 162^Tand P. espejianaATCC 29659^Tdepended on the type and the concentration of carbohydrate substrates in the nutrient media. The molar proportion between rhamnose, glucose, and galactose (ca. 1 : 0.3 : 2) in the PS of P. elyakoviiKMM 162^Twas almost the same in the media lacking carbohydrates or containing glucose or galactose at a concentration of 1 g/l. At the same time, the molar proportion between fucose, glucose, galactose, galactosamine, and glucosamine (ca. 1 : 1 : 1 : 2 : 0.5) in the PS of P. espejianaATCC 29659^Tdepended on the presence and the concentration of carbohydrate substrates in the medium. A high concentration of glucose in the medium (30 g/l) brought about a rise in the content of glucose in PSs (9-fold for the PS of P. elyakoviiKMM 162^Tand 4.6-fold for the PS of P. espejianaATCC 29659^T) and led to a decrease in the content of other carbohydrates. The cultivation of these two strains at a lactose concentration of 30 g/l resulted in their PSs containing glucose and galactose in about equal proportions (ca. 1 : 1 in the case of P. espejianaATCC 29659^Tand ca. 2.1 : 1.7 in the case of P. elyakoviiKMM 162^T).     

Effects of different batches of iodine on properties of I-hFSH and characteristics of radioligand-receptor assays

Radioiodination of highly purified human follicle-stimulating hormone (hFSH) (4000 IU/mg) was performed every other week for 23 weeks using 2 mCi carrier free Na ¹²⁵I (Amersham Corp., 15 mCi/μg I₂) in the presence of lactoperoxidase. Incorporation of ¹²⁵I into hFSH was determined by the method of [7.]Biochem. J. 89, 114). Hormone binding was studied in vitro under steady-state conditions (16 h, 20°C) using different calf testis membrane preparations having similar receptor characteristics. Each ¹²⁵I-hFSH preparation was characterized for maximum bindability, specific activity of bindable radioligand as determined by self-displacement analysis, and by determination of K_a and R_t. Incorporation of ¹²⁵I into FSH was relatively constant over the large number of experiments (62.4 ± 6.4 μCi/μg; n = 23). By comparison, however, specific radioactivity of the receptor bindable fraction of ¹²⁵I-hFSH was related to the lot of ¹²⁵I utilized, and was significantly (P ≤ 0.01) lower and more variable (28.7 ± 10.5 μCi/μg). Maximum bindability of ¹²⁵I-hFSH was not correlated to specific activity (r = 0.06) but was negatively correlated to hFSH ¹²⁵I incorporation (r = −0.47; P ≤ 0.05). These observations demonstrate the need to assess the quality of each batch of radioligand before undertaking radioligand-receptor assays and suggest that differences in Na¹²⁵I lots affect specific radioactivity of the radioligand and its receptor binding characteristics.     

Estimation of genome sizes of hyperthermophiles

Genomes of various hyperthermophilic and extremely thermophilic prokaryotes were analyzed with respect to size, physical organization, and 16S rDNA copy number. Our results show that all the genomes are circular, and they are in the size range of 1.6–1.8 Mb for Pyrodictium abyssi, Methanococcus igneus, Pyrobaculum aerophilum, Archaeoglobus fulgidus, Archaeoglobus lithotrophicus, and Archaeoglobus profundus (the two bacteria Fervidobacterium islandicum and Thermosipho africanus possess genomes of 1.5-Mb size). A systematic study of all validly described species of the order Sulfolobales revealed the existence of two classes of genome size for these archaea, correlating with phylogenetic analyses. The Metallosphaera–Acidianus group, plus Sulfolobus metallicus, have genomes of ca. 1.9 Mb; the other members of the order Sulfolobales group possess genomes >2.7 Mb. The special case of Stygiolobus azoricus is discussed. Received: August 10, 1997 / Accepted: January 1, 1998     

Seasonal study of the fungal biota of the fur of dogs

During a one year period, 944 dogs from the Municipal kennel of Barcelona were examined to detect animals with suspected dermatophytosis. Only a few animals (1.8%) presented skin lesions but none of them had dermatophytosis. A representative number of dogs without visible skin lesions (n=172), selected at random, were used to carry out a seasonal study of the mycobiota of their fur. Fifteen isolates belonging to the genera Microsporum and Trichophyton were isolated from 14 of the 172 (8.1%) dogs without lesions. The identity of these fungi was Microsporum gypseum (6/15), Trichophyton terrestre (4/15), M. canis (2/15), M. cookei (2/15) and Trichophyton ajelloi (1/15) (one strain each of M. gypseum and T. ajelloi were isolated from one dog). Species of Penicillium (% prevalence=89.5%), Alternaria (86.6%), Cladosporium (84.9%), Aspergillus (77.3%), Scopulariopsis (65.7%) and Chrysosporium (64.5%) were the most prevalent. No significant differences in the fungal biota were observed with respect to age, gender, hair length or between mixed and pure breed dogs. A large number of isolates, including species belonging to the genera Beauveria, Chrysosporium, Malbranchea and Scopulariopsis, that macroscopically and/or microscopically resemble dermatophytes and may be mistaken for them, produced a red color change in Dermatophyte Test Medium. No significant seasonal difference was detected among the isolates belonging to the the most frequently encountered genera, with the exception of Scopulariopsis (higher in summer and autumn) and Chrysosporium (higher in summer). Species from other genera, with lower occurrence also presented significant differences in their seasonal distribution. Arthrinium, Aureobasidium, Chaetomium and Phoma spp. presented maximum prevalence peaks in spring, Fusarium, Paecilomyces, Phoma and Rhizopus spp. in summer and Geotrichum and Mucor spp. in autumn. The Microsporum and Trichophyton species were more frequently isolated in summer.     

Comparison of the profiles of non-glycosylated triterpenoids from leaves of plants of selected species of genus Dioscorea

Remarkable qualitative and quantitative differences in non-glycosylated triterpenoid profiles of twelve Dioscorea spp. leaves were demonstrated with the use of GC–MS/FID analysis. The total content of tetracyclic triterpenoids and their esters ranged from 397 μg/g of dry leaf weight in D. bulbifera to 762 μg/g d.w. in D. discolor and 777 μg/g d.w. in D. alata. Three main phytosterols, i.e. campesterol (1), sitosterol (2) and stigmasterol (3) were found in extracts from all analyzed species in total amount ranging from 316 μg/g in D. bulbifera to 676 μg/g of dry leaf weight in D. hispida, with either sitosterol (2) or stigmasterol (3) as predominant in the profile. Extracts from D. hispida and D. purpurea leaves were distinguished from the others by particularly high amount of campesterol (1). In the majority of the species, except for D. caucasica, other tetracyclic triterpenoids were found: cycloartanol (4), 24-methylenecycloartanol (5) and cycloeucalenol (6). Less common steroids, stigmastan-3-en-6β-ol (7) and ergosta-7,22-dien-3β-ol (8) were detected in D. japonica. The significant content (992 μg/g) of pentacyclic triterpenoids of ursane, oleanane, taraxastane and taraxerene (friedooleanane)-type carbon skeletons, i.e. α-amyrin (9), β-amyrin (10), taraxasterol (11) and taraxerol (12), respectively, was found in D. caucasica. The obtained results supplement the knowledge of biochemical diversity of Dioscorea genus.     

Stimulation by interferon of induction of differentiation of human promyelocytic leukemia cells

F₁-ATPase was isolated from yeast $\text{S.}\text{cerevisiae}$. The constituent subunits 1 and 2 were purified by gel permeation chromatography, and their amino acid compositions determined. Both subunits have a similar composition except for $\text{1}\text{2}$ cystine, methionine, leucine, histidine, and tryptophan. When F₁ is treated for three hours with 5′-p-[³H]fluorosulfonylbenzoyl adenosine in dimethylsulfoxide, 90% of the activity is lost. Disc gel electrophoresis of the modified complex showed that over 90% of the label was associated with subunit 2. A labelled peptide from a $\text{S.}\text{aureus}$ digest of subunit 2 was isolated and sequenced. It had the following amino acid sequence: His-Try¹-Asp-Val-Ala-Ser-Lys-Val-Gln-Glu, whereby Tyr¹ is the modified amino acid residue. This sequence shows homology to other sequences obtained from maize, beef heart, and $\text{E.}\text{coli}$ F₁-ATPases.     

Preparation and regeneration of protoplasts of three fungi of Boletaceae

Protoplasts of three fungi of Boletaceae,Suillus luteus, S. grevillei, andBoletinus cavipes, were prepared with yields of 45, 8.0, and 1.8×10⁷/g fresh mycelia under the optimal conditions, respectively. Nucleate protoplasts accounted for 42% of the whole preparation ofS. luteus and 32% of that ofS. grevillei, and 21% of the nucleate protoplasts ofS. luteus and 35% of those ofS. grevillei possesed two nuclei. Regeneration efficiency of protoplasts was 0.4% forS. luteus and 0.05% forS. grevillei. The regeneration ofB. cavipes protoplasts was also confirmed. Optimal conditions for regeneration were determined. Addition of gellan gum instead of agar to the medium and activated charcoal treatment of agar medium increased the regeneration efficiency significantly.     

Comparison of Vanadium-Rich Activity of Three Species Fungi of Basidiomycetes

A comparison of vanadium-rich activity of three species fungi of Basidiomycetes, Ganoderma lucidum, Coprinus comatus, and Grifola frondosa, was studied. By fermentation and atomic absorption spectroscopy analysis, the biomass of G. lucidum and G. frondosa declined rapidly when the concentration of vanadium exceeded 0.3% but the biomass of C. comatus did not decline rapidly until the concentration of vanadium exceeded 0.4% and the content of vanadium accumulated in the mycelia was 3529.3 μg/g. After the mice were administered (intragastrically) with vanadium-rich C. comatus, the blood glucose of alloxan-induced hyperglycemic mice was decreased (p < 0.05) and the body weight of the alloxan-induced hyperglycemic mice was increased gradually. Thus, we selected C. comatus to absorb vanadium and chose 0.4% as the optimal concentration of vanadium for the pharmacological works.