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。     

不同因子对长春花愈伤组织中药用成分积累的影响

 目的 研究继代次数、继代龄(继代组织块的年龄)、生长素、金属离子等因子的改变对长春花愈伤组织生长及吲哚生物碱类药用成分积累的影响。方法 测定不同因子的影响下,长春花愈伤组织的鲜重,并对吲哚总碱进行提取、含量测定和比较。结果 随继代次数的增加,长春花愈伤组织中吲哚总碱含量有所下降;20 d继代龄最利于组织生长;2,4-二氯苯氧乙酸的存在对吲哚总碱的积累有一定的抑制作用,但却有利于愈伤组织生长,两种生长素组合比单一的生长素更利于组织生长,也较利于吲哚总碱积累;钙离子浓度为220 mg·L^-1,锌离子浓度为8.6哗·L^-1时,对提高吲哚总碱产量最有效。结论 继代次数、继代龄、生长素、金属离子对长春花愈伤组织生长及吲哚生物碱类药用成分的积累有较大影响。     

Correlation between wing bone microstructure and different flight styles: The case of the griffon vulture (gyps fulvus) and greater flamingo (phoenicopterus roseus)

Flying is the main means of locomotion for most avian species, and it requires a series of adaptations of the skeleton and of feather distribution on the wing. Flight type is directly associated with the mechanical constraints during flight, which condition both the morphology and microscopic structure of the bones. Three primary flight styles are adopted by avian species: flapping, gliding, and soaring, with different loads among the main wing bones. The purpose of this study was to evaluate the cross-sectional microstructure of the most important skeletal wing bones, humerus, radius, ulna, and carpometacarpus, in griffon vultures (Gyps fulvus) and greater flamingos (Phoenicopterus roseus). These two species show a flapping and soaring flight style, respectively. Densitometry, morphology, and laminarity index were assessed from the main bones of the wing of 10 griffon vultures and 10 flamingos. Regarding bone mineral content, griffon vultures generally displayed a higher mineral density than flamingos. Regarding the morphology of the crucial wing bones involved in flight, while a very slightly longer humerus was observed in the radius and ulna of flamingos, the ulna in griffons was clearly longer than other bones. The laminarity index was significantly higher in griffons. The results of the present study highlight how the mechanics of different types of flight may affect the biomechanical properties of the wing bones most engaged during flight.     

长春花叶片中吲哚生物碱增产的研究

目的探讨乙烯利、乙酰水杨酸、色氨酸3种试剂对长春花植株中药用成分积累的影响。方法用不同质量浓度的3种试剂溶液分别处理温室盆栽长春花植株,2d后,测定叶片中的总吲哚生物碱含量,并采用RP-HPLC法测定长春碱和长春质碱的含量。结果3种试剂处理后吲哚总碱、长春碱、长春质碱的积累均有明显的提高,并确定了最适的处理质量浓度。结论3种试剂对长春花植物中吲哚生物碱的产生有明显的促进作用。     

高速逆流色谱法分离纯化长春花中长春新碱

目的采用高速逆流色谱(HSCCC)分离纯化长春花中长春新碱,建立相关工艺条件,为工业生产提供参考数据。方法采用高速逆流色谱仪,氯仿-甲醇-0.3mol.L-1 HCl(4∶3∶2)溶剂体系,上相为固定相,下相为流动相,流动相流速为2.0mL.min-1,紫外检测波长为280nm,主机转速为800r.min-1,恒温循环温度为28℃。结果 HPLC检测HSCCC纯化后长春新碱质量分数为79.7%。结论高速逆流色谱纯化长春花中长春新碱容量大、分离纯度和效率高,避免了有机溶剂的大量使用,适用于工业生产高纯度产品。     

长春花组织培养研究进展

长春花含有100多种吲哚类生物碱,具有抗肿瘤、降血压等多种生物活性,有较高的药用价值,但是含量偏低。长春花组织培养可从提高繁殖系数、调控次生代谢产物的累积等来提高长春花生物碱类成分含量。本文就外植体和培养基的选择、药用成分累积的影响因素、悬浮细胞培养应用、影响毛状根因素和基因工程技术应用等方面,综述了长春花组织培养的主要研究进展,为进一步开发利用长春花药物资源提供参考。     

Severe Bone Marrow Depression Induced by an Anticancer Herb Cantharanthus Roseus

We report a 67‐yr‐old woman with hepatitis C–related liver cirrhosis and hepatoma who had developed severe bone marrow suppression after taking Cantharanthus roseus as an alternative anticancer treatment. The patient developed severe pancytopenia with initial presentations of vomiting, diarrhea, oral ulcer, and fever about 1 week after taking 5‐days' course of Cantharanthus roseus. Bone marrow biopsy showed autolysis, which indicated massive necrosis of the hematopoietic cells. There was no malignant cell infiltration. The patient also had severe gastrointestinal disturbances, bacteremia, urinary tract infection, and impaired renal and liver function. Supportive care with broad‐spectrum antibiotics, granulocyte colony‐stimulating factor, repeated blood transfusions, and albumin supplement was given. She recovered and was discharged after 48 days hospitalization. Coadministration of Cantharanthus roseus and cisapride was noted, and these two drugs are both substrates of cytochrome P450 3A4 enzymes (CYP 3A4). Because the vinca alkaloids are extensively metabolized by the liver cytochrome P450 enzymes, poor hepatic function and drug‐herb interaction might predispose the patient to develop the bone marrow toxicity. This case report demonstrated possible effect of oral dose of vinca alkaloids and also hinted that all the substrates and inhibitors of CYP 3A4 have propensity to interfere with metabolism of vinca alkaloids.     

Synthesis of silver nanoparticles using leaves of Catharanthus roseus Linn. G. Don and their antiplasmodial activities

Objective

To develop a novel approach for the green synthesis of silver nanoparticles using aqueous leaves extracts of Catharanthus roseus (C. roseus) Linn. G. Don which has been proven active against malaria parasite Plasmodium falciparum (P. falciparum).

Methods

Characterizations were determined by using ultraviolet-visible (UV-Vis) spectrophotometry, scanning electron microscopy (SEM), energy dispersive X-ray and X-ray diffraction.

Results

SEM showed the formation of silver nanoparticles with an average size of 35–55 nm. X-ray diffraction analysis showed that the particles were crystalline in nature with face centred cubic structure of the bulk silver with the broad peaks at 32.4, 46.4 and 28.0.

Conclusions

It can be concluded that the leaves of C. roseus can be good source for synthesis of silver nanoparticle which shows antiplasmodial activity against P. falciparum. The important outcome of the study will be the development of value added products from medicinal plants C. roseus for biomedical and nanotechnology based industries.     

长春花细胞C_(20)hi生物碱超量合成及激素自养的分子机制探讨

以长春花激素自养型细胞株C20hi及其母株C20D为材料,构建抑制性差减cDNA文库,斑点杂交筛选后对阳性克隆进行测序,共得到104个不同的差异表达基因片段。将这些序列在GenBank中进行BLAST比对分析,其中28个与已知基因具有很高的同源性。对部分基因的分析发现长春花细胞C20hi中g10h-like、CrPS(Catharanthus roseus protein S)、HyPRP(hybrid prolin-rich protein)、PNAE(polyneuridine aldehyde ester-ase)-like等基因的差异表达可能与该细胞株生物碱超量合成有关;而细胞色素P45071D和CR5基因的差异表达则可能与其激素自养有关。此外,培养基中2,4-D的缺失引起的TDC等酶活性的增强也可能是C20hi细胞生物碱超量合成及激素自养的原因之一。     

长春花内生真菌的分离及其发酵产生药用成分的初步研究

首次报道从长春花 Catharanthus roseus (L.) G. Don茎的韧皮部中分离出尖孢镰刀菌Fusariumoxysporum, 为该植物的一种内生真菌, 并用 TLC和 HPLC对该菌的97CG₃菌株培养物进行了分析, 初步结果表明该真菌能产生抗癌药长春新碱成分。