龙葵不同采收期及不同部位中澳洲茄碱与澳洲茄边碱的含量分析

目的比较不同采收期、不同部位龙葵中澳洲茄碱、澳洲茄边碱的含量。方法采用高效液相色谱法测定不同采收期、不同部位龙葵中澳洲茄碱、澳洲茄边碱的含量。结果 6~8月采集的样品中澳洲茄碱、澳洲茄边碱的含量以7月份最高。龙葵全草、果实、茎、叶、果柄中,果实含量最高。结论不同采收期、不同部位龙葵中澳洲茄碱、澳洲茄边碱的含量存在明显差异,应注意采收季节和药用部位对龙葵药材质量的影响。     

假烟叶树叶中澳洲茄碱和澳洲茄边碱的HPLC-ELSD测定

目的建立高效液相色谱-蒸发光散射检测(HPLC-ELSD)测定假烟叶树叶中澳洲茄碱和澳洲边茄碱的方法。方法通过方法学要素考察检测方法的可行性。结果以0.1%三氟乙酸水溶液和乙腈作为流动相,流速1.0 mL/min,柱温35℃;ELSD漂移管温度70℃,雾化管温度30℃,载气压力35psi,经梯度洗脱,澳洲茄碱和澳洲茄边碱达到良好分离;样品用质量50倍的1%甲酸甲醇在60℃超声提取30min,提取液直接进样。澳洲茄碱对照品在0.62～6.2g线性关系良好,加标回收率为100.95%;澳洲茄边碱在0.2～7.5g线性关系良好,加标回收率为95.34%。测定样品中澳洲茄碱含量为1.19mg/g;澳洲茄边碱含量为1.06 mg/g。结论该方法能准确快速地测定假烟叶中澳洲茄碱和澳洲茄边碱的含量。     

香瓜茄的化学成分分析

目的:对茄科Solanaceae植物甘肃武威香瓜茄Solanum muicatum成熟果实的化学成分进行分离纯化研究,并对分离得到的化合物进行化学成分鉴定。方法:采用95%的乙醇对香瓜茄干果进行加热提取,并依次用石油醚,乙酸乙酯,正丁醇进行萃取,然后采用薄层色谱法,正相硅胶柱色谱法,LH-20羟丙基葡聚糖凝胶色谱法和制备高效液相色谱法等方法对香瓜茄95%乙醇提取物的乙酸乙酯萃取部位进行分离纯化,并依据化合物的理化性质并通过核磁共振、质谱等现代波谱技术对化合物的化学结构进行鉴定。结果:最终从香瓜茄中分离纯化得到7个化合物,其中3个生物碱皂苷类化合物,2个黄酮类化合物及2个有机酸类化合物,分别鉴定为澳洲茄碱(solasonine 1),澳洲茄边碱(solamargine 2),澳洲茄胺(solasodine 3),槲皮素(quercetin 4),柚皮素(naringenin 5),阿魏酸(ferulic acid 6),熊果酸(ursone 7)。结论:以上7种化合物均为首次从该植物果实中分离纯化得到,该研究为综合开发及寻找天然植物性抗炎、抗肿瘤活性物质提供一定的化学依据和基础。     

龙葵药材中澳洲茄碱、澳洲茄边碱的含量测定

目的:寻找可以作为评价龙葵药材质量的指标成分,提高龙葵药材的质量标准。方法:采用柱层析方法分离化学成分,应用光谱法进行结构鉴定,用高效液相色谱法测定其在龙葵药材不同部位的含量。结果:从龙葵药材中分离并鉴定了2个含量较高的甾体类生物碱苷澳洲茄碱(solasonine)和澳洲茄边碱(solamar-gine);建立了龙葵药材中澳洲茄碱、澳洲茄边碱的HPLC含量测定方法。结论:这两种生物碱在龙葵药材中含量较高,可用于龙葵药材的质量控制。     

龙葵中澳洲茄碱含量的动态规律

目的：通过研究龙葵不同产地、生长周期及药用部位中澳洲茄碱的动态规律,为进一步优化龙葵的采收期、药用部位及栽培提供依据。方法：采用HPLC,AgilentTC-C18色谱柱（250 mm×4.6 mm）;以乙腈-0.5%磷酸水溶液作为流动相,梯度洗脱;检测波长为203 nm;体积流量：0.9 mL·min-1;柱温为30 ℃。结果：龙葵药材中澳洲茄碱含量因产地不同差异很大。龙葵地上部分澳洲茄碱含量的高低主要取决于所结果实的多少,以盛果期含量最高;不同成熟程度果实中澳洲茄碱含量的规律为：成熟果实＜青果＜幼果。结论：在龙葵的生长过程中,龙葵中澳洲茄碱的含量随着其生长发育进程而发生变化。根据龙葵不同生长阶段所含澳洲茄碱在各器官中的含量变化,龙葵的最佳采收期为盛果期,最佳药用部位为幼果和青果。     

不同产地的龙葵果中澳洲茄碱、澳洲茄边碱的含量测定比较研究

目的比较不同采收期、同部位龙葵中澳洲茄碱、澳洲茄边碱的含量。方法采用高效液相色谱法测定不同产地龙葵中澳洲茄碱、澳洲茄边碱的含量。结果以吉林产的鲜龙葵果含含澳洲茄碱、澳洲茄边碱的量最高,安徽,辽宁,河北等地的龙葵干果的含量也较高。结论不同产地的龙葵果中鲜果与干果里面所含有澳洲茄碱以及澳洲茄边碱的含量存在显著性差异,应注意采时的产地和龙葵果的新鲜程度对龙葵药材质量的影响。     

龙葵果保鲜技术对澳洲茄碱、澳洲茄边碱含量的影响

目的:研究龙葵果保鲜技术对其抗肿瘤成分澳洲茄碱、澳洲茄边碱含量的影响。方法:采用独特的龙葵果保鲜技术保存龙葵鲜果,并采用高效液相色谱法对龙葵干果和保鲜处理的鲜果中澳洲茄碱、澳洲茄边碱进行含量测定,比较保鲜技术对其抗肿瘤有效成分含量的影响。结果:经保鲜技术处理的龙葵鲜果中澳洲茄碱和澳洲茄边碱的含量要比龙葵干果的含量高80%以上。结论:龙葵果保鲜技术可以明显提高龙葵药材中抗肿瘤有效成分的含量,值得推广使用。     

无梗五加果实乙醇提取物抗肿瘤有效部位的研究

目的确定无梗五加果实乙醇提取物抗肿瘤的有效部位。方法建立小鼠肝癌H22肿瘤模型,检测无梗五加果实80%乙醇提取物的不同溶剂萃取物:石油醚萃取物、二氯甲烷萃取物、正丁醇萃取物以及剩余水溶性成分,对小鼠体内肿瘤生长的抑制作用以及对脾脏指数、胸腺指数、细胞因子水平的影响。结果无梗五加果实乙醇提取物的不同溶剂萃取物中石油醚萃取物高剂量组、二氯甲烷萃取物高剂量组以及水溶性成分低剂量组对小鼠移植性实体瘤H22有明显抑制作用,抑瘤率分别为41.88%、42.55%、44.83%。与模型组比较,无梗五加果实各萃取物对于荷瘤小鼠的胸腺指数、脾脏指数以及血清中IL-2、IL-6、TNF-α的水平均有不同程度的提高。结论无梗五加果实乙醇提取物中石油醚层、二氯甲烷层、水溶性成分层是无梗五加果实乙醇提取物抗肿瘤的有效部位。     

龙葵果的神经药理活性

龙葵Solanum nigrum L.是常用草药,其果实在墨西哥用作健神药,人们亦用它治疗神经官能症。从该植物中已分离鉴定澳洲茄次碱(solasodine)、澳洲茄碱(solasonine)、茄啶(solanidine)等化合物,但其神经药理作用尚未阐明。作者研究了龙葵果乙醇提取物的神经药理活性。将龙葵果碾碎并制成乙醇提取物,收率为7.9％。用大鼠观察其对戊巴比妥睡眠时     

大黄不同饮片大孔树脂分离部位的提取物总量及其HPLC比较研究

目的:通过对大黄不同饮片大孔树脂分离部位的提取物总量及其HPLC比较,确定饮片炮制前后成分变化明显的组分,探索导致大黄药性变化的药效物质基础。方法:以75%乙醇提取大黄各种饮片,提取物经大孔树脂D101分离,分别制备成水、20%乙醇、50%乙醇和95%乙醇部位,以HPLC对各部位进行比较。结果:饮片总提取物重量由高到低为生片、酒片>醋片>熟片>炭片。酒片、醋片及炭片均为50%乙醇洗脱物最多,生片和熟片均为H₂O洗脱物最多,但生片20%和50%乙醇洗脱物的重量基本一致,而熟片则呈依次递减趋势。各饮片4个洗脱部位中均为95%乙醇洗脱部位的洗脱物最少。HPLC比较结果显示,各组分分离良好,各饮片间的20%和50%及95%乙醇洗脱部位色谱峰变化显著。结论:随着炮制条件的加剧,各饮片化学成分的组成及含量发生了较明显的变化,提取物重量及HPLC比较结果提示,20%,50%及95%乙醇洗脱部位化学成分的变化可能是导致大黄不同饮片药性变化的物质基础。     

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.     

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.     

Screening of the topical anti-inflammatory activity of the bark of Acacia cornigera Willdenow, Byrsonima crassifolia Kunth, Sweetia panamensis Yakovlev and the leaves of Sphagneticola trilobata Hitchcock

Ethnopharmacological relevance

An investigation of topical anti-inflammatory activity was undertaken on plants used in Central America traditional medicine.

Aim of study

Four herbal drugs used in the folk medicine of Central America to treat inflammatory skin affections (Acacia cornigera bark, Byrsonima crassifolia bark, Sphagneticola trilobata leaves and Sweetia panamensis bark) were evaluated for their topical anti-inflammatory activity.

Materials and methods

Petroleum ether, chloroform and methanol extracts were obtained for herbal medicines and then extracts were tested on Croton oil-induced ear dermatitis model in mice.

Results

Almost all the extracts reduced the Croton oil-induced ear dermatitis in mice and the chloroform ones showed the highest activity, with ID₅₀ (dose giving 50% oedema inhibition) values ranging from 112 μg/cm² (Byrsonima crassifolia) to 183 μg/cm² (Sphagneticola trilobata). As reference, ID₅₀ of the non-steroidal anti-inflammatory drug indomethacin was 93 μg/cm².

Conclusions

Lipophilic extracts from these species can be regarded as potential sources of anti-inflammatory principles.     

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。     

Antifungal activity of Brazilian medicinal plants involved in popular treatment of mycoses

A survey of medicinal plants used to treat common mycoses was done in the Curituba district, Sergipe State, Brazil. One hundred inhabitants were interviewed by health agents and traditional healers. Four different plants were the most cited (more than 50% of the citations): Ziziphus joazeiro, Caesalpinia pyramidalis, Bumelia sartorum and Hymenea courbaril. The aqueous extracts obtained following traditional methods and using different parts of these plants, were submitted to drop agar diffusion tests for primary antimicrobial screening. Only the water infusion extract of Ziziphus joazeiro and Caesalpinea pyramidalis presented a significant antifungal activity against Trichophyton rubrum, Candida guilliermondii, Candida albicans, Cryptococcus neoformans and Fonsecaea pedrosoi, when compared to the antifungal agent amphotericin B. The minimal inhibitory concentration (MIC) of the bioactive extracts was evaluated by the microdilution method. Best activity with a MIC of 6.5 microg/ml for both extracts was observed against Trichophyton rubrum and Candida guilliermondii. Ziziphus joazeiro and Caesalpinea pyramidalis extracts presented also low acute toxicity in murine models. The present study validates the folk use of these plant extracts and indicates that they can be effective potential candidates for the development of new strategies to treat fungal infections.     

Experimental study on anti-inflammatory activity of a TCM recipe consisting of the supercritical fluid CO2 extract of Chrysanthemum indicum, Patchouli Oil and Zedoary Turmeric Oil in vivo

Ethnopharmacological relevance

Chrysanthemum indicum (Compositae) Linné, Pogostemon cablin (Blanco) Benth and Curcuma wenyujin (Zingiberaceae) Y. H. Chen et C. Ling are three of the extensively used herbal remedies among traditional Chinese medicines for the purpose of anti-inflammation. A traditional Chinese medicine (TCM) recipe named CPZ consisting extracts of the above three herbs, has shown noteworthy anti-influenza activity, which is closely related to its anti-inflammatory feature.

Aim of this study

To investigated the anti-inflammtory activity of CPZ in vivo for a further exploration of the recipe's anti-inflammatory properties.

Materials and methods

The anti-inflammatory property of CPZ on acute inflammation was evaluated by inflammatory models of dimethylbenzene (DMB)-induced ear vasodilatation and acetic acid-induced capillary permeability enhancement in mice, as well as the carrageenan-induced paw edema rat model, in which inflammation-related cytokine including prostaglandin E₂ (PGE₂), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and nitric oxide (NO) in the edematous paw tissue were determined by enzyme linked immunosorbent assay (ELISA). Moreover, effect of CPZ on chronic inflammation was observed through granuloma formation in rats subjected to cotton pellet implantation.

Results

CPZ (340, 170, and 85 mg/kg for mice, p.o.) not only decreased the DMB-induced ear vasodilatation but also attenuated capillary permeability under acetic acid challenge in mice. And the significant inhibition on carrageenan-induced paw edema was observed. Further more, the ELISA results showed that CPZ (170, 85, and 42.5 mg/kg for rats, p.o.) could up-regulate the level of IL-1β in the edema paw tissue of rats significantly while down-regulate that of PGE₂, but no apparent effect on TNF-α or NO was observed in the test. Besides, CPZ had a certain degree of restraining effect on the cotton pellet-induced granuloma formation in rats and the highest dose of 170 mg/kg even showed a significant suppression on it.

Conclusion

The above results indicated that CPZ possessed a potent anti-inflammatory activity, which is indicated to be closely associated with its regulation on IL-1β and PGE₂ thereby mediating the inflammatory response acting at an appropriate level.     

Regulation effects on abnormal glucose and lipid metabolism of TZQ-F,a new kind of Traditional Chinese Medicine

Aim of the study

Based on the recipe of the traditional anti-diabetic formula TZQ, we developed TZQ-F, a new formula including 8 fractions isolated from Red Paeony root, Mulberry leaf, Lotus leaf, Danshen root and Hawthorn leaf with a good quality assurance. The study was aimed at fraction preparation and effects of the fractions on abnormal glucose and lipid metabolism.

Materials and methods

The active fractions were obtained by macroporous resin, ion-exchange resin and polyamide resin column chromatographies. HPLC analyses were used for quality control. In vitro mechanism study included DPPH radical scavenging, AGEs formation inhibition, α-glucosidase inhibition and lipase inhibition, and rats on high-fat diet were used for in vivo study.

Results

In vitro mechanism study showed that among the 8 fractions, three of them had inhibition effects on intestinal disaccharase, three with inhibition effects on lipase, and five with effects of free radical scavenging. In vivo study showed that after 4 weeks of treatment, TZQ-F significantly decreased the levels of serum total cholesterol, TG, glucose, LDL-C and HDL-C in rats on high-fat diet. Consistent with the in vitro and in vivo results, histology study demonstrated that TZQ-F alleviated hepatic steatosis induced by high-fat diet.

Conclusions

TZQ-F possesses the potential regulation effects on abnormal glucose and lipid metabolism.     

常用藏药“蒂达”(藏茵陈)基原物种药用合理性及资源利用价值评价

目的:对“蒂达”各品种不同基原植物药用合理性及其资源利用价值作出评价.方法:采用HPLC指纹图谱共有特征峰、结合SPSS聚类分析方法,评价“蒂达”不同品种基原植物的成分组成相似性.结果:“蒂达”不同品种涉及的来源有龙胆科獐牙菜属、花锚属、扁蕾属及肋柱花属共15种1变种33个样品,指纹图谱全谱相似度较差;以10个共有峰成分的相对面积聚类,其基原植物可分别聚为4个组;上述4个属的属间成分组成差异较大;其中獐牙菜属不同种间成分组成变化较大,而其他各属种间成分组成相似度较高.结论:上述龙胆科4属植物均含有具有保肝利胆生物活性的环烯醚萜类、(山)酮类及三萜类成分,藏医学将其归为“蒂达”一类药材使用具有一定的物质基础;鉴于“蒂达”类药材不同基原各品种间在成分组成上差异较大,分别制定各品种的HPLC指纹图谱,是鉴别和控制各品种质量的有效手段,可有效控制各基原品种质量.     

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。     

虫草属EST-SSR标记系统的建立研究

目的:通过球孢虫草、蛹虫草EST设计EST-SSR引物,建立虫草属EST-SSR标记系统.方法:从NCBI公共数据库下载获得虫草EST,利用Sequece Seiners 1.2软件去除冗余序列并设计引物,进行PAGE电泳.结果:通过去除EST总序列中低质量的和冗余的序列后,得到全长为2 953 173 bp的4 556条无冗余球孢虫草EST.从中发掘出718个EST-SSR,分布于616条EST中,出现频率是15.8％.平均分布频率是每4 096 bp出现1个,三核苷酸重复序列有419个,是出现最多的重复类型.蛹虫草EST去冗余后得到1 363条无冗余EST,共含有1 117个EST-SSR,出现频率为81.95％,出现最多的重复类型是A核苷酸重复.根据球孢虫草EST-SSR序列,设计合成50对引物,有扩增产物的引物为34对,占总设计引物数的68％.根据蛹虫草EST-SSR,设计合成40对引物,有扩增产物的引物为39对,占总设计引物数的97.5％.基于SSR标记进行聚类分析,7种虫草无性型均能分开,且分为4支.结论:虫草属EST-SSR出现频率较高、类型较丰富、多态性潜能较高,具有较高的利用价值.球孢虫草和蛹虫草EST开发的SSR标记在虫草属有较好的转移性与通用性,可以很好的应用于虫草种间遗传关系的研究.应用虫草物种EST建立分子标记是一条简便而又有效的途径.