姜辣素拮抗掌叶半夏毒针晶刺激性毒性的机制研究

为探索姜辣素拮抗掌叶半夏毒针晶刺激性毒性的机制,该研究采用掌叶半夏毒针晶诱导的小鼠腹腔炎症模型,观察姜辣素对小鼠腹腔炎症渗出液中炎症介质PGE2的影响;采用大鼠腹腔巨噬细胞体外培养模型,以上清液中TNF-α和IL-1β为指标,研究姜辣素拮抗掌叶半夏毒针晶的致炎作用;采用扫描电镜法观察姜辣素拮抗掌叶半夏毒针晶刺激后巨噬细胞表面形态的改变;采用巨噬细胞-中性粒细胞共培养模型,考察姜辣素拮抗掌叶半夏毒针晶对中性粒细胞迁移的影响。结果显示,姜辣素可以显著抑制掌叶半夏毒针晶所致的小鼠腹腔渗出液中PGE2的生成;姜辣素可以显著抑制掌叶半夏毒针晶诱导巨噬细胞释放炎症因子,并具有一定的量效关系;扫描电镜显示,姜辣素可以显著抑制巨噬细胞吞噬掌叶半夏毒针晶及细胞膜破损,并能显著抑制掌叶半夏毒针晶诱导的中性粒细胞迁移。姜辣素拮抗掌叶半夏毒针晶刺激性毒性的机制可能是抑制了包括巨噬细胞激活、炎症因子释放、中性粒细胞迁移聚集的致炎毒性。     

半夏及掌叶半夏毒针晶中共性毒蛋白的研究

目的:寻找天南星科半夏属有毒中药半夏及掌叶半夏毒性物质基础毒针晶中的共有蛋白,研究刺激性相关蛋白的致炎效应。方法:采用溶液内双酶解方法将半夏与掌叶半夏毒针晶中的蛋白降解为混合肽段,采用液相色谱与串联质谱偶联分析降解后的肽段,将质谱分析数据与BIOWORKS软件搜索的NCBI绿色植物Viridiplantae蛋白库比对进行蛋白质检索分析。采用聚丙烯酰胺凝胶电泳对半夏、掌叶半夏毒针晶蛋白进行分析,同时采用胶内酶解后液相色谱与串联质谱偶联的方法,分析并鉴定约13kDa蛋白。提取分离半夏中的凝集素蛋白,并采用中性粒细胞迁移试验和测定腹腔渗出液中PGE2的含量,考察毒针晶及凝集素的炎症刺激性效应。结果:半夏及掌叶半夏毒针晶中均含约13kDa的蛋白条带,此条带经鉴定为凝集素类蛋白,且此13kDa蛋白条带为毒针晶中的主要蛋白之一,凝集素类蛋白可诱导中性粒细胞向大鼠腹腔迁移,可显著增加小鼠腹腔渗出液中PGE2的含量。结论:天南星科有毒中药半夏及掌叶半夏毒针晶蛋白中均含有凝集素类蛋白,且此类凝集素蛋白具有显著的致炎作用,是半夏与掌叶半夏毒针晶中的共性毒蛋白。     

掌叶半夏(虎掌南星)矾制前后致炎毒性比较研究

目的: 探索掌叶半夏致炎毒性及白矾炮制对其致炎作用的影响。 方法: 自掌叶半夏鲜品中分离毒针晶及掌叶半夏凝集素蛋白;采用整体动物及细胞水平模型,以炎症介质为指标,考察掌叶半夏毒性成分毒针晶和凝集素蛋白的致炎效应,及矾制对致炎作用的影响。 结果: 腹腔注射掌叶半夏毒针晶混悬液可使小鼠腹腔渗出液中炎症介质PGE₂,NO含量显著增加,矾制后生品及毒针晶诱导的PGE₂,NO释放显著下降;掌叶半夏凝集素蛋白刺激小鼠巨噬细胞后,在一定时间内可导致剂量依赖性的炎症因子TNF-α及IL-6含量显著增加。矾制后凝集素蛋白诱导的小鼠腹腔渗出液中PGE₂含量显著下降。 结论: 掌叶半夏的刺激性毒性在机体表现为炎症反应,其所含的毒针晶及凝集素蛋白是其毒性成分,均可导致显著的炎症介质释放。矾制可显著降低掌叶半夏毒性成分的致炎毒性。     

半夏凝集素蛋白与半夏毒针晶毒性的相关性研究

目的:研究半夏凝集素蛋白与半夏毒针晶毒性的相关性,揭示半夏产生毒性的机制。方法:采用大鼠腹腔炎症模型,以腹腔渗出液中PGE2含量为指标,研究半夏凝集素致炎作用的量-毒、时-毒曲线;采用家兔眼结膜刺激模型,以眼结膜的病理切片观察半夏凝集素与半夏毒针晶刺激性的相关性。结果:半夏凝集素可引起大鼠腹腔渗出液中PGE2、蛋白含量显著提高,呈剂量依赖性;半夏凝集素可显著加重半夏毒针晶对家兔眼结膜的刺激性,且随着半夏凝集浓度的升高,刺激性显著增强,但单给药半夏凝集素对家兔眼结膜无刺激性。结论:半夏凝集素蛋白具强烈的致炎作用。半夏产生强烈炎症刺激的机制是半夏刺入机体,凝集素蛋白随针晶进入机体组织,诱导炎症反应发生而产生刺激性。     

黄柏抗痛风活性部位筛选及其对腹膜炎小鼠的保护作用

目的 研究黄柏不同萃取部位抗痛风作用，筛选有效部位并初步探究其作用机制。方法 采用单钠尿酸盐(MSU)诱导小鼠腹腔巨噬细胞及骨髓来源巨噬细胞(BMDM)建立细胞痛风模型，经黄柏不同极性部位给药后，ELISA法检测肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)分泌，从而筛选出活性部位，最后研究活性部位在小鼠体内抗痛风作用及相关机制。结果 黄柏醇提液各萃取部位中，正丁醇部位能降低细胞IL-1β水平和巨噬细胞的吞噬能力(P<0.01),但对TNF-α无明显影响(P>0.05)。与模型组比较，正丁醇部位各剂量组小鼠腹腔灌洗液IL-1β、IL-6水平和中性粒细胞募集程度均降低(P<0.05,P<0.01),其中高剂量组效果最明显，但各给药组TNF-α水平无明显变化(P>0.05)。结论 正丁醇部位为黄柏抗痛风活性部位，其抗痛风机制可能是通过降低细胞吞噬MSU晶体的能力，从而减少IL-1β、IL-6等细胞因子释放，进而减少中性粒细胞募集程度，改善痛风症状。     

藏药忍冬果对小鼠腹腔巨噬细胞吞噬功能及分泌细胞因子的影响

目的:研究藏药忍冬果水提物(FLM)对小鼠腹腔巨噬细胞吞噬功能及分泌细胞因子的影响。方法:运用吞噬中性红试验观察FLM对小鼠腹腔巨噬细胞吞噬功能的影响,用小鼠腹腔巨噬细胞分泌物对小鼠胸腺细胞增殖及对L929细胞杀伤作用的影响分别测定IL-1及TNF-α的活性,用RT-PCR技术观察药物对小鼠腹腔巨噬细胞TNF-αmRNA和IFN-γmRNA表达的影响。结果:FLM在1～100μg.mL^-1剂量内对小鼠腹腔巨噬细胞吞噬功能有明显促进作用,同时能诱导巨噬细胞分泌IL-1及TNF-α,并促进细胞因子TNF-αmRNA和IFN-γmRNA的表达。结论:FLM对小鼠腹腔巨噬细胞吞噬功能及分泌细胞因子都有促进作用。     

杨梅素对小鼠脾淋巴细胞和腹腔巨噬细胞增殖的影响

目的:研究杨梅素体外对小鼠脾脏淋巴细胞增殖及腹腔巨噬细胞活化的影响。方法:取健康小鼠,MTT法检测杨梅素对小鼠脾淋巴细胞增殖的作用;中性红法测腹腔巨噬细胞吞噬活性;Griess试剂法测NO释放量;Elisa法测TNF-α、IL-1β和IL-4的含量。结果:杨梅素对未活化及丝裂原(Con A、LPS)活化的淋巴细胞增殖均有明显促进作用;可剂量依赖性地增强小鼠腹腔巨噬细胞吞噬中性红能力和促进NO释放;促进RAW264.7细胞TNF-α、IL-1β产生,抑制IL-4的释放。结论:杨梅素可能通过促进小鼠脾淋巴细胞增殖和提高巨噬细胞吞噬及分泌功能,提高机体免疫功能。     

半夏及其毒针晶刺激性毒性的稳定性研究

目的:考察半夏毒针晶刺激性毒性的稳定性及针晶蛋白对针晶刺激性毒性的贡献,为进一步揭示半夏毒针晶产生刺激性毒性的机制提供依据。方法:不同溶剂及混合蛋白酶液浸泡处理半夏及其毒针晶,用家兔眼结膜刺激性模型考察半夏及其所含针晶的刺激性变化,用电镜扫描法观察毒针晶晶体结构变化,采用垂直凝胶电泳(SDS-PAGE)试验分析蛋白酶液浸泡前后针晶蛋白的条带变化。结果:半夏毒针晶在甲醇、乙醇浸泡24 h内刺激性逐渐消失,半夏生品在75%乙醇溶液润制2 d后刺激性降低,扫描电镜显示经过甲醇、乙醇处理的毒针晶变得易于断碎,针尖锈蚀。混合蛋白酶液浸泡3 min后的半夏毒针晶刺激性急剧下降,SDS-PAGE图谱显示蛋白酶液浸泡后的针晶蛋白被酶解。结论:半夏针晶蛋白对针晶刺激性有贡献。甲醇、乙醇浸泡可使半夏毒针晶刺激性下降,提示可能是由于极性有机溶剂作用于针晶蛋白,逐渐破坏针晶蛋白的功能。     

半夏毒针晶的致炎作用研究

目的探讨半夏毒针晶引起的刺激性炎症的机体表现及对相关炎症介质的影响。方法采用小鼠毛细管通透性实验、小鼠腹腔炎症实验及大鼠足跖肿胀实验,考察不同剂量半夏毒针晶的致炎效应及其量-效关系,并测定相关炎症介质的量。结果半夏毒针晶混悬液可使小鼠腹腔毛细血管通透性增加,腹腔渗出液中炎症介质前列腺素E2(PGE2)、一氧化氮(NO)、丙二醛(MDA)的量增加;亦可引起大鼠足跖肿胀,并在一定剂量范围内表现为典型的量-效关系,还可使大鼠致炎足跖中炎症介质PGE2、环氧合酶-2(COX-2)的量显著增加。结论半夏毒针晶刺激性毒性在机体内表现为炎症反应。     

多糖类药物作用的受体及信号转导机制的研究进展

β-葡聚糖通过补体3型受体(CR3)增强巨噬细胞、NK细胞、中性粒细胞的杀伤作用;多数多糖可通过Toll样受体4(TLR4)或TLR2激活和增强巨噬细胞的吞噬功能,或诱导巨噬细胞合成包括肿瘤坏死因子α(TNF-α)在内的多种细胞因子;也有报道β-葡聚糖的主要受体是树突状细胞相关C型凝集素-1(dectin-1),而且介导了巨噬细胞的生物功能。     

Effect of feeding Mucuna pruriens on helminth parasite infestation in lambs

Ethnopharmacological relevance

Mucuna pruriens is a tropical legume anecdotally reputed to have anthelmintic properties. This study was conducted to examine the validity of such claims.

Aim of the study

The aim of this study was to determine if ingestion of Mucuna seeds reduces helminth parasite infestation in lambs.

Materials and methods

Thirty-six Dorper × Katahdin ram lambs were assigned to three treatments, a cottonseed meal based control diet, a diet in which Mucuna replaced cottonseed meal and the control diet with levamisole (7.5 mg/kg body weight) administration. All diets were isonitrogenous and isocaloric. The 12 lambs in each treatment were assigned randomly to 4 pens, each containing 3 lambs. Lambs were trickle infected three times per week by gavage with infectious Haemonchus contortus larvae (2000 larvae/lamb) for 3 weeks.

Results

Levamisole treatment decreased fecal egg counts by 87% and abomasal worm counts by 83%. Mucuna intake did not statistically affect fecal egg counts or abomasal worm counts, though numerical (P > 0.10) reductions of 7.4% and 18.1%, respectively were evident. Anemia indicators, feed intake, and lamb growth were unaffected by treatment.

Conclusions

Levamisole reduced the Haemonchus parasite burden in lambs significantly but feeding Mucuna reduced the burden by levels unlikely to eliminate the clinical effects of parasitism.     

Investigation of plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts

Aim of the study

To investigate the activities of the 217 plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts.

Materials and methods

Plant extracts were prepared by the method of maceration using solvents of different polarities. The growth inhibition of chloroquine-resistant Plasmodium falciparum strain (FcB1) was determined by measuring the radioactivity of the tritiated hypoxanthine incorporated. Activity against Leishmania (Leishmania) chagasi and Trypanosoma cruzi was measured by the MTT colorimetric assay. The antifungal tests were carried out by using the CLSI method. The active extracts were tested also by cytotoxicity assay using NIH-3T3 cells of mammalian fibroblasts.

Results

Two hundred and seventeen extracts of plants were tested against Plasmodium falciparum. The eleven active extracts, belonging to eight plant species were evaluated against L. (L.) chagasi, Trypanosoma cruzi, yeasts and in NIH-3T3 cells. The results found in these biological models are consistent with the ethnopharmacological data of these plants. The ethyl acetate extract of Diospyros hispida root showed IC₅₀ values of 1 μg/mL against Plasmodium falciparum. This extract demonstrated no toxicity against mammalian cells, resulting in a significant selectivity index (SI) of 435.8. The dichloromethane extract of Calophyllum brasiliense root wood was active against Cryptococcus gattii LMGO 01 with MIC of 1.95 μg/mL; and Candida albicans ATCC 10231 and Candida krusei LMGO 174, both with MIC of 7.81 μg/mL. The same extract was also active against Plasmodium falciparum and L. (L.) chagasi with IC₅₀ of 6.7 and 27.6 μg/mL respectively. The ethyl acetate extract of Spiranthera odoratissima leaves was active against Cryptococcus gattii LMGO 01 with MIC of 31.25 μg/mL, and against Plasmodium falciparum with IC₅₀ of 9.2 μg/mL and Trypanosoma cruzi with IC₅₀ of 56.3 μg/mL.

Conclusion

The active extracts for protozoans and human pathogenic yeasts are considered promising to continue the search for the identification and development of leading compounds.     

α-Glucosidase inhibitory activity of selected Philippine plants

Ethnopharmacological relevance

Antidesma bunius Spreng. (Phyllantaceae), Averrhoa bilimbi L. (Oxalidaceae), Biophytum sensitivum (L.) DC. (Oxalidaceae), Ceriops tagal (Perr.) C.B. Rob. (Rhizophoraceae), Kyllinga monocephala Rottb. (Cyperaceae), and Rhizophora mucronata Lam. (Rhizophoraceae) are used as remedies to control diabetes. In the present study, these plants were screened for their potential α-glucosidase inhibitory activity.

Materials and methods

The 80% aqueous ethanolic extracts were screened for their α-glucosidase enzyme inhibitory activity using yeast alpha glucosidase enzyme.

Results

Except for A. bilimbi with IC₅₀ at 519.86±3.07, all manifested a significant enzyme inhibitory activity. R. mucronata manifested the highest activity with IC₅₀ at 0.08±1.82 μg mL⁻¹, followed by C. tagal with IC₅₀ at 0.85±1.46 μg mL⁻¹ and B. sensitivum with IC₅₀ at 2.24±1.58 μg mL⁻¹.

Conclusion

This is the first report on the α-glucosidase inhibitory effect of the six Philippine plants; thus, partly defining the mechanism on why these medicinal plants possess antidiabetic properties.     

European medicinal polypores – A modern view on traditional uses

Ethnopharmacological relevance

In particular five polypore species, i.e. Laetiporus sulphureus, Fomes fomentarius, Fomitopsis pinicola, Piptoporus betulinus, and Laricifomes officinalis, have been widely used in central European folk medicines for the treatment of various diseases, e.g. dysmenorrhoea, haemorrhoids, bladder disorders, pyretic diseases, treatment of coughs, cancer, and rheumatism. Prehistoric artefacts going back to over 5000 years underline the long tradition of using polypores for various applications ranging from food or tinder material to medicinal–spiritual uses as witnessed by two polypore species found among items of Ötzi, the Iceman. The present paper reviews the traditional uses, phytochemistry, and biological activity of the five mentioned polypores.

Materials and methods

All available information on the selected polypore taxa used in traditional folk medicine was collected through evaluation of literature in libraries and searches in online databases using SciFinder and Web of Knowledge.

Results

Mycochemical studies report the presence of many primary (e.g. polysaccharides) and secondary metabolites (e.g. triterpenes). Crude extracts and isolated compounds show a wide spectrum of biological properties, such as anti-inflammatory, cytotoxic, and antimicrobial activities.

Conclusions

The investigated polypores possess a longstanding ethnomycological tradition in Europe. Here, we compile biological results which highlight their therapeutic value. Moreover, this work provides a solid base for further investigations on a molecular level, both compound- and target-wise.     

真菌的中药干预研究近5年进展

近年来真菌感染率逐年上升,传统抗真菌药物易产生耐药性,而中药在防治真菌感染方面具有一定的优势。本文就近5年来中药对白念珠菌、皮肤癣菌、曲霉菌、马拉色菌、串珠镰孢菌、申克孢子丝菌、新生隐球菌及真菌生物膜的干预研究进展进行综述。     

厚朴与凹叶厚朴群体遗传学研究

目的:对厚朴与凹叶厚朴的群体遗传学进行研究,为中药厚朴的质量控制提供分子生药学方面的依据。方法:对厚朴与凹叶厚朴15个居群应用2个叶绿体基因间序列psbA-trnH和trnL-trnF进行PCR扩增并测序,计算厚朴与凹叶厚朴单倍型频率,用程序HaploNst分析遗传多样性和遗传结构,应用TCS version 1.13软件构建单倍型网状进化树。结果:厚朴与凹叶厚朴均无特有单倍型存在,但单倍型频率存在显著差异,已开始出现遗传分化的趋势,NST略大于GST。结论:厚朴与凹叶厚朴在遗传上已出现遗传分化的趋势,但尚未完全分化成彼此独立的单系。     

Antiplasmodial activity of extracts from seven medicinal plants used in malaria treatment in Cameroon

Aim of the study

In a search for new plant-derived biologically active compounds against malaria parasites, we have carried out an ethnopharmacological study to evaluate the susceptibility of cultured Plasmodium falciparum to extracts and fractions from seven Cameroonian medicinal plants used in malaria treatment. We have also explored the inhibition of the Plasmodium falciparum cysteine protease Falcipain-2.

Materials and methods

Plant materials were extracted by maceration in organic solvents, and subsequently partitioned or fractionated to afford test fractions. The susceptibility of erythrocytes and the W2 strain of Plasmodium falciparum to plant extracts was evaluated in culture. In addition, the ability of annonaceous extracts to inhibit recombinant cysteine protease Falcipain-2 was also assessed.

Results and discussion

The extracts showed no toxicity against erythrocytes. The majority of plant extracts were highly active against Plasmodium falciparumin vitro, with IC₅₀ values lower than 5 μg/ml. Annonaceous extracts (acetogenin-rich fractions and interface precipitates) exhibited the highest potency. Some of these extracts exhibited modest inhibition of Falcipain-2.

Conclusion

These results support continued investigation of components of traditional medicines as potential new antimalarial agents.     

中国石斛属植物文献计量研究

石斛是珍稀濒危中药材,目前正处于快速发展阶段。为全面了解我国石斛属植物研究的历史和发展现状,作者以1954～2010年"中国知网中国学术期刊网络出版总库"收录的石斛研究文献为依据,采用文献计量学的原理和方法,对我国石斛属植物研究文献从文献年代分布、期刊分布与被引频率、主题分布、研究对象分布、作者与研究机构分布等方面进行了统计与分析。结果表明,我国石斛研究明显分为起步(2个)、停滞、平稳发展、快速上升5个阶段;期刊分布存在离散性与集中性并存的现象,已形成核心期刊研究群,并以《中国中药杂志》、《中草药》和《陕西中医》为代表;研究主题广泛涉及临床与药理、组织培养与种苗繁育、成分分析等多个领域,已经形成比较稳定的研究机构和团队,但研究对象差异显著,以铁皮石斛、金钗石斛和霍山石斛最为集中。我国石斛属植物的研究已取得显著成果,但种植产业发展缓慢,供需矛盾突出,预计种苗繁育与人工种植、产品开发、化学与药理等方面是未来的研究热点,其文献报道仍将进一步上升。     

列当属、肉苁蓉属和草苁蓉属植物传统药物学调查

通过野外资源调查、整理各大标本馆标本原始记录和查阅文献记载等方法,系统整理、总结、归纳了列当属、肉苁蓉属和草苁蓉属民族药用植物种类、功效及民间使用情况,结果表明列当属6种药用植物在4个少数民族间作为7种民族药应用,草苁蓉属2种药用植物在8个少数民族间作为10种民族药应用,肉苁蓉属2种药用植物在3个少数民族间作为3种民族药应用,且同种药用植物常作不同民族药;发现3属植物的传统疗效主要集中在补肾壮阳、止血和止痛3个方面,并且该传统疗效与现代药理研究结果基本吻合。因此深入研究植物种类丰富的列当属植物资源对缓解肉苁蓉植物资源匮乏局面和扩大药源具有积极意义。     

牛耳朵和黄花牛耳朵的显微和化学鉴别

目的:为苦苣苔科唇柱苣苔属植物牛耳朵和黄花牛耳朵的鉴定和分类提供解剖学和化学依据。方法:采用石蜡制片法和水合氯醛透化法对2种药用植物的根状茎和叶横切面和粉末特征进行研究,应用光学显微镜观察显微结构。采用HPLC-UV法进行化学鉴别。结果:牛耳朵和黄花牛耳朵显微特征无明显区别,但是化学特征有明显的差异。结论:化学诞生特征鉴别方法可以作为2种药用植物的鉴定方法。