两面针化学成分、药理活性及抗肿瘤机制研究进展

本文主要结合近几年的国内外研究报道,综述了中国传统中草药-两面针的化学成分、药理活性及临床疗效等方面的研究进展;并结合作者所在课题组的研究成果,重点介绍了两面针生物碱及其金属配合物的抗肿瘤活性研究。     

Folin-Ciocalteu比色法测定花椒中多酚含量的研究

研究了以没食子酸为标准品,采用Folin-Ciocalteu比色法,测定花椒多酚含量的适宜条件。结果表明:当Folin-Ciocalteu试剂与10%Na2CO3溶液之比为1∶2(V∶V),Folin-Ciocalteu试剂体积为4 mL,反应温度为25℃,反应时间为120 min时,多酚含量在1~6μg/mL范围内与吸光值呈良好的线性关系。测定了4种花椒中的多酚含量,四川青花椒多酚含量最高。     

Optimized genetic transformation of Zanthoxylum zanthoxyloides by Agrobacterium rhizogenes and the production of chelerythrine and skimmiamine in hairy root cultures

Zanthoxylum zanthoxyloides is an endangered African tree producing numerous bioactive substances including antileukemic and antisickling agents. Here, the potential of Z. zanthoxyloides hairy root cultures was tested for the production of bioactive substances with limited natural resources. The efficiency of Agrobacterium rhizogenes LBA9402‐mediated transformation of leaf material was evaluated using different techniques. An optimal transformation frequency of 77% was obtained after 11 days by inoculating A. rhizogenes directly onto the central vein of 14‐week‐old leaves followed by a co‐cultivation period of 3 days. Different treatments in immersion mode (manual wounding, acetosyringone, CaCl₂, ultrasonication) never exceeded these results. A maximum growth rate of 0.37 cm/day was determined during the exponential phase. Liquid chromatography‐diode array detection analysis showed the presence of skimmiamine, sesamine, chelerythrine, and chelerythrine derivatives in Z. zanthoxyloides hairy root lines. The maximum production of skimmiamine and chelerythrine in 28‐day‐old hairy root cultures was 45 ± 2 and 107 ± 4 mg/100 g dry weight, respectively. The present results highlight the potential of Z. zanthoxyloides hairy root cultures for the sustainable production of skimmiamine and chelerythrine.     

蚬壳花椒ISSR-PCR反应体系的建立及优化

通过单因素试验及正交设计方法对影响蚬壳花椒ISSR-PCR扩增的主要因素(模板DNA、Mg2+的浓度、引物、dNTPs,TaqDNA聚合酶的用量以及退火温度)进行优化,以建立蚬壳花椒ISSR-PCR反应的最佳体系。结果表明:最佳反应体系(20μL)为模板DNA 60ng、MgCl2 2.5mmol/L、dNTPs 0.15mmol/L、引物0.6μmol/L、TaqDNA聚合酶2.4U。在此基础上,从100条引物中筛选出18条扩增稳定、多态性好的ISSR引物并经过10份蚬壳花椒种质检验,证明该体系具有扩增条带清晰、稳定、重复性好等优点。该反应体系的建立为蚬壳花椒种质资源分类、遗传多样性分析提供了更客观可靠的方法。     

Isolation and Characterization of 7S Protein-I of Phaseolus angularis (Adzuki bean)

The major storage protein of Phaseolus angularis, 7S protein-I, was purified by gel filtration on Sephadex G–200 and ion exchange chromatography on DEAE-cellulose. The purified 7S protein-I was homogeneous on disc electrophoresis, isoelectric focusing and ultracentrifugation. The sedimentation coefficient (, w) and Stokes radius of 7S protein-I were estimated to be 7.5 S and 48 Å, respectively. The molecular weight of 7S protein-I was calculated to be 150,000 ± 15,000 from these values. Polyacrylamide gel electrophoresis of 7S protein-I in the presence of sodium dodecyl sulfate showed one main (55,000 ± 3000 daltons) and two minor protein bands (28,000 ± 1400 and 25,000 ± 1300 daltons). 7S protein-I contained large amounts of glutamic acid and aspartic acid but no cysteine and low amounts of methionine. Carbohydrate analysis of 7S protein-I revealed the presence of 5.0% of neutral sugars and 0.5 % of amino sugars. Circular dichroism measurements indicated that this protein is a β-form rich protein.     

中国花椒属(广义)叶结构研究

对国产花椒属32种4变种叶结构特征进行了研究。结果发现,花椒属叶脉均属于羽状脉弓形脉序类型;叶齿具全缘、具齿或钝齿3种类型;二级叶脉有分支或不分支;三级叶脉有分支、网状和及顶3种类型;多数种类叶齿无腺点,仅4种叶齿有腺点。叶结构研究结果支持花椒亚属和崖椒亚属的亚属等级,它们共同组成广义的花椒属,并支持花椒属为芸香科中比较原始类群的观点;但不支持Engler将花椒亚属和崖椒亚属独立成属的分类处理。基于重要外部形态学特征和叶结构特征,重新对花椒属下两个亚属的检索表进行了修订。     

Benzophenanthridine alkaloids from Zanthoxylum nitidum (Roxb.) DC, and their analgesic and anti-inflammatory activities

Seven benzophenanthridine alkaloids, 1-7, were isolated from the roots of Zanthoxylum nitidum. Among them, two novel alkaloids, named (R)-8-[(R)-1-hydroxyethyl]dihydrochelerythrine (1) and 8-methoxynorchelerythrine (2), were structurally identified as new compounds on the basis of the spectroscopic analysis. Bioactivity evaluation showed that nitidine (3), dihydrochelerythrine (4), oxyavicine (5), 8-methoxychelerythrine (6), and 8-hydroxydihydrochelerythrine (7) exhibit comparable analgesic and anti-inflammatory effects as hydrocortisone.     

超临界二氧化碳萃取技术在花椒麻味成分分离中的应用研究

要在超临界迷流体萃取花椒过程中,使用乙醇作为夹带剂,将花椒中的麻味成分如山椒素等萃取分离出来,探讨了萃取压力、温度以及夹带荆纯度等因素对萃取效果的影响。得到最佳萃取工艺：夹带剂加入量为5％,纯度为80％,萃取压力为30Mpa。温度为50℃,时间为1h。     

超临界CO2萃取技术应用于花椒油树脂分离工艺的研究

本文研究了超临界迷萃取花椒油树脂的工艺,探讨了萃取压力、温度和时间等因素对花椒萃取率的影响,通过正交实验法确定了花椒超临界迷流体萃取的最佳工艺：花椒原料粒度为60日,萃取条件为压力32Mpa、温度55℃、时间1h。     

两面针中的化学成分

研究两面针Zanthaxylum nitidum(Roxb．)DC根部乙醇提取物中的化学成分。采用硅胶柱层析方法进行分离和纯化,通过波谱分析鉴定分离得到的化合物的结构。共鉴定7种香豆素成分及2种其它类型成分,其中6种香豆素前文已作过报道。本文主要介绍三种化学成分即β-谷甾醇(1),5-methoxymarmesin(2),β-香树素(3)。其中5-methoxymarmesin为首次从该植物中分离得到的化合物。