黄花蒿与其近缘种化学成分的FTIR和GC-MS鉴定与分析

黄花蒿是一种治疗痢疾的特效中药,植物体中含有丰富的精油,但其应用和生产中常有种类混杂现象,严重影响了黄花蒿为原料的药材质量。为实现黄花蒿药材快速鉴定与评价,该研究利用FTIR技术和GCMS分别对黄花蒿及其近缘种叶片原药材及挥发油成分进行了检测和鉴定。结果表明:挥发油以黄花蒿含量最高(1.86%),其次是南牡蒿、茵陈蒿、青蒿、牡蒿和艾蒿。FTIR分析结果表明,黄花蒿及其近缘种一维图谱相似,酰胺类、芳香类以及萜类化合物种类较多且含量丰富;二阶导数图谱中,黄花蒿青蒿素成分振动吸收明显增强,可以明显将黄花蒿与其混淆中区分开。GC-MS分析显示,黄花蒿与其近缘种的挥发油成分中共检测出17个共有峰,28种化学成分,均含有较高樟脑、á-杜松烯、Crocetane、植烷、2,4-二叔丁基苯酚,但不同种间成分含量差异很大,植烷在黄花蒿中含量明显高于其它近缘种,龙脑成分只能在黄花蒿叶片中检测出,然而á-雪松烯在青蒿、南牡蒿、茵陈蒿均较高,而在黄花蒿,艾蒿,牡蒿中含量均较低。最后通过聚类分析探讨了黄花蒿与其近缘种挥发油成分差异性,6种材料明显聚为2类。其中,黄花蒿与牡蒿、艾蒿聚为一类,青蒿与茵陈蒿和南牡蒿聚为一类。该研究结果为黄花蒿药材的真伪鉴别及其药材质量评价提供了快速而有效的分析手段。     

栽培与野生黄花蒿中化学组分的FTIR表征及青蒿素含量比较分析

本研究运用FTIR技术,对栽培和野生黄花蒿不同部位化学成分的红外光谱特征进行指认,并比较分析两者不同部位青蒿素含量差异.结果显示:酰胺、芳香类物质均以栽培黄花蒿茎和叶片中多;可溶性多糖和糖苷类物质,栽培与野生黄花蒿茎中含量相近,但是,叶片中含量以栽培黄花蒿高;纤维素类物质,栽培茎少于野生茎,而两种黄花蒿叶片的纤维素含量相近.与青蒿素标准品比较,栽培和野生黄花蒿不同部位青蒿素含量有一定差异,其中栽培叶最高,其次是野生叶,栽培茎的含量最低.所以,运用FTIR技术可以快速判断栽培和野生型黄花蒿主要化学成分的差异,本研究对黄花蒿的引种驯化和良种选育工作有一定指导意义.     

青蒿素提取研究进展

从黄花蒿的选择、青蒿素提取溶剂的选择、黄花蒿微波预处理、超临界CO_2萃取技术、微波辅助提取、超声波强化石油醚提取等方面叙述青蒿素提取的研究进展,并将这几种方法进行比较,为工业上大规模生产青蒿素提供借鉴。     

野生与栽培黄花蒿净光合速率对光强和CO2浓度的响应

比较了相同种源的野生和栽培黄花蒿(Artemisia annua L.)净光合速率对光强和CO2浓度的响应特性。结果表明,野生和栽培黄花蒿的光饱和点(LSP)分别为1 183和1 564μmol m-2s-1,光补偿点(LCP)为17和18μmol m-2s-1,最大净光合速率(Pmax)为18.78和22.38μmol m-2s-1,表观量子效率(AQY)为0.08和0.075μmol m-2s-1,表明黄花蒿的光合能力强,能够利用很高的光强,且对弱光的适应性也较强。栽培黄花蒿的Pmax、LSP和最大羧化速率(Vcmax)显著高于野生黄花蒿,两者的LCP、不包括光下呼吸的CO2补偿点、AQY、光下呼吸速率和最大电子传递速率(Jmax)差异不显著。强光下栽培黄花蒿主要通过提高Vcmax和Jmax等来增强光合能力,强的光合能力有利于黄花蒿的生长,因此在人工栽培黄花蒿的过程中应选择阳光充足的开阔生境。     

提取青蒿素的原植物

青蒿素是新型抗疟药的有效成分,而提取青蒿素的原植物则是一种习见杂草黄花蒿。介绍了黄花蒿的形态特征、生境及分布特点,以及资源利用现状,分析了我国中药青蒿的渊源,以及可能包含的植物种类,并指出了青蒿与黄花蒿之间的区别。     

广西黄花蒿类型调查研究

本文报道广西黄花蒿类型调查结果。１）生长在石山上的黄花蒿类型植株长势比生长在平地、路边、土坡、房前屋后的类型差,且产量也较低。２）同一分布区内,生长在石山的黄花蒿类型,有效成分（青蒿素）含量明显高于生长在平地、路边的类型,前者比后者含量提高００３％～０２８％。     

不同土壤环境对黄花蒿生长和青蒿素含量的影响研究

通过田间小区试验,比较研究了施肥与不施肥条件下,4种土壤环境（沙土、旱地土、水稻土和棕色石灰土）对黄花蒿的生长、生物量分配和青蒿素含量的影响。结果表明：黄花蒿对土壤养分的适应性较强,在沙土、旱地土、水稻土和石灰土上均能生长发育,养分水平低时,分配更多的生物量到根,根生物量分数和根/冠比增大;养分水平高时,分配更多的生物量到叶,叶生物量分数增加。黄花蒿的生长和青蒿素含量显著受土壤养分的影响,不施肥时,石灰土和水稻土栽培黄花蒿的株高、地径、总生物量、叶生物量和青蒿素含量显著大于旱地土,而旱地土又显著大于沙土。但在施肥条件下,以上参数不同土壤间无显著差异,且显著高于不施肥。因此,只要根据土壤养分状况合理施肥,黄花蒿在不同养分土壤栽培均能获得较高的青蒿素产量。     

武陵山区黄花蒿生长发育规律研究

为了解武陵山区黄花蒿(Artemisia annua)的生长特性,对其生长发育规律进行了研究。结果表明,黄花蒿的生长发育可划分为苗期、分枝初期、分枝盛期、蕾期、花期、种子成熟期。叶面积在分枝盛期增长最快,叶面积指数随着枝叶的生长呈直线上升。种子成熟期叶面积比呈总体降低的趋势。净同化率变化较小,分别在苗期和花期出现了两个小高峰。相对生长率变化较大,作物生长率在苗期和种子成熟期较低,分枝盛期较高。整个生长期干物质积累规律呈S曲线。6月下旬至8月下旬,黄花蒿生长进入分枝盛期,干物质积累速率加快,叶片产量迅速增加,此时期是黄花蒿叶片增产的关键时期,应注重氮磷钾肥的平衡施用。     

干旱胁迫对黄土高原4种蒿属植物叶形态解剖学特征的影响

以黄土高原演替初期阶段的猪毛蒿(Artemisia scoparia Waldst.et Kit.)、茭蒿(Artemisia giraldii Pamp.)、铁杆蒿(Artemisia gmelinii Web.ex Stechm.)3种优势种和1种常见种黄花蒿(Artemisia annua Linn.)为供试材料,采用盆栽试验与称重控水法,将土壤含水量分别控制在田间最大持水量的75%(适宜水分对照)、55%(中度干旱)和35%(重度干旱),研究干旱胁迫对菊科蒿属植物叶片形态解剖学的影响,揭示植物叶片生态适应机制,为生态恢复过程中植被演替的规律提供理论依据。于植物营养生长末期选取代表性的叶片,分别采用电子扫描显微镜、电子透射显微镜和光学显微镜对叶表皮特征、亚细胞显微特征和组织结构进行观察。结果表明:干旱胁迫下,这4种植物表现为不同程度的叶片增厚、栅栏组织厚度、叶片紧实度和角质层厚度增加;3种旱生型植物猪毛蒿、茭蒿和铁杆蒿的叶片下表皮具有浓密的表皮毛,有利于防止水分过度蒸腾,而中生型黄花蒿叶片无此功能;铁杆蒿和黄花蒿上表皮毛稀少,不利于保水和防止强光照,干旱胁迫下依靠降低上表皮气孔密度和叶细胞叶绿体数目来适应环境;茭蒿和猪毛蒿叶绿体形态相似,具有较厚的基粒和浓密的基粒片层,强光下容易引起氧化损伤,猪毛蒿通过浓密的上表皮毛而茭蒿通过较强的抗氧化机制防止光能过剩,两者在重度干旱下均产生大量脂质体。4种蒿属植物中黄花蒿的叶片解剖学特征受到干旱的影响变化最大,铁杆蒿和猪毛蒿次之,茭蒿最弱,3种优势种的干旱适应性显著强于黄花蒿,干旱下叶片解剖学的结论与干旱耐受性指标结果一致。     

黄花蒿花序精油化学成分初步分析

黄花蒿(Artemisia annua L.)系菊科蒿属植物,为常用中草药.黄花蒿花序精油具有独特香气,青香中略带花香,头香有甜、凉气息。为充分利用国内这一丰富野生植物资源,我们对其精油成分进行了 GC/MS/DS 分析和主成分的层析分离、鉴定。实验结果如下。实验部分1.精油的制取将采集于长春地区的新鲜黄花蒿花序进行水蒸汽蒸馏,接收采用二次冷凝系统。所     

促进黄花蒿发根青蒿素合成的内生真菌诱导子的制备

应用酸解法对黄花蒿(ArtemisiaannuaL.)内生胶孢炭疽菌(Colletotrichumgloeosporioides)菌丝体进行提取,在黄花蒿发根培养系统中比较了各制备提取物的青蒿素诱导活性。活性提取物经过SephadexG25层析后,部分纯化的内生菌寡糖提取物(MW<2500)可显著促进发根青蒿素的合成,培养23d的发根经诱导子(0.4mg/mL)处理4d后,青蒿素产量可达13.51mg/L,比同期对照产量提高51.63%,诱导作用与诱导子浓度、作用时间相关。内生菌寡糖诱导子的制备和使用,在青蒿素生物技术生产研究中为首次应用。     

青蒿倍半萜合酶(环化酶)研究进展

青蒿素是从中药青蒿中分离得到的抗疟有效单体,是含有过氧基团的新型倍半萜内酯化合物,是目前世界上最有效的疟疾治疗药物。青蒿素的生物合成途径属于类异戊二烯代谢途径中的倍半萜类分支途径,倍半萜合酶是该途径的关键酶之一,目前已从青蒿中克隆了多个倍半萜合酶基因。综述了青蒿中已克隆的几种倍半萜合酶基因的研究进展。     

Yield enhancement strategies for artemisinin production by suspension cultures of Artemisia annua

Artemisinin, isolated from the shrub-Artemisia annua, is a sesquiterpene lactone used to treat multi-drug resistant strains of falciparum malaria. It is also effective against a wide variety of cancers such as leukemia and colon cancer. To counter the present low content in leaves and uneconomical chemical synthesis, alternate ways to produce artemisinin have been sought. But this compound remains elusive in cell cultures of A. annua despite the extensive studies undertaken. This work reports the first successful approach for production of artemisinin by cell cultures of Indian variety of A. annua. In the present study, an integrated yield enhancement strategy, developed by addition of selected precursor (mevalonic acid lactone) and elicitor (methyl jasmonate) at optimized concentrations, resulted in 15.2g/l biomass and 110.2mg/l artemisinin, which was 5.93 times higher in productivity in comparison to control cultures.     

State of the art of the production of the antimalarial compound artemisinin in plants

For more than three centuries we have relied on the extracts of the bark of Cinchona species to treat malaria. Now, it seems we may be changing to the leaves of a Chinese weed, Artemisia annua, and its active compound artemisinin. Artemisinin-derived drugs have been proved particularly effective treatments for severe malaria, even for multi-drug-resistant malaria. However, this promising antimalarial compound remains expensive and is hardly available on a global scale. Therefore, many research groups have directed their investigations toward the enhancement of artemisinin production in A. annua cell cultures or whole plants in order to overproduce artemisinin or one of its precursors. This article provides a brief review of the state of art of the different aspects in A. annua research.     

Methyl jasmonate and miconazole differently affect arteminisin production and gene expression in Artemisia annua suspension cultures

Artemisia annua L. is a herb traditionally used for treatment of fevers. The glandular trichomes of this plant accumulate, although at low levels, artemisinin, which is highly effective against malaria. Due to the great importance of this compound, many efforts have been made to improve knowledge on artemisinin production both in plants and in cell cultures. In this study, A. annua suspension cultures were established in order to investigate the effects of methyl jasmonate (MeJA) and miconazole on artemisinin biosynthesis. Twenty-two micro molar MeJA induced a three-fold increase of artemisinin production in around 30 min; while 200 μm miconazole induced a 2.5-fold increase of artemisinin production after 24 h, but had severe effects on cell viability. The influence of these treatments on expression of biosynthetic genes was also investigated. MeJA induced up-regulation of CYP71AV1, while miconazole induced up-regulation of CPR and DBR2.     

Effects of Overexpression of the Endogenous Farnesyl Diphosphate Synthase on the Artemisinin Content in Artemisia annua L.

Artemisinin is a novel effective antimalarial drug extracted from the medicinal plant Artemisia annua L. Owing to the tight market and low yield of artemisinin, there is great interest in enhancing the production of artemisinin. In the present study, farnesyi diphosphate synthase （FPS） was overexpressed in high-yield A. annua to Increase the artemisinin content. The FPS activity in transgenic A. ennue was twoto threefold greater than that In non-transgenic A. annua. The highest artemisinin content in transgenic A. annua was approximately 0.9% （dry weight）, which was 34.4% higher than that in non-transgenic A. annua. The results demonstrate the regulatory role of FPS in artemisinin biosynthesis.     

TLC-densitometric analysis of artemisinin for the rapid screening of high-producing plantlets of Artemisia annua L

A simple TLC-densitometric technique has been developed for the rapid and accurate analysis of artemisinin in a large number of Artemisia annua plantlets cultured in vitro. This new analytical method is based on the structural conversion of artemisinin on a silica gel layer by ammonia vapour to form 10-azadesoxyartemisinin, a chromophore-containing compound (lambdamax 320 nm) that can be detected by UV-based TLC densitometry. The TLC system was evaluated quantitatively in terms of product stability, precision, accuracy and calibration. Good linearity was obtained in the range of 0.01-0.12 microg artemisinin. The technique appeared to be accurate and sensitive as compared with the complicated pre-column reaction-HPLC technique. Among 90 samples of A. annua plantlets, the artemisinin content in the leaves appeared to be highly variable, ranging from 0.02 to 0.67% w/w dry weight. These results demonstrate that densitometric TLC can be a cheap and simple technique for the accurate screening of high-artemisinin-producing plants.     

Dihydroartemisinin regulates the immune system by promotion of CD8~+ T lymphocytes and suppression of B cell responses

Artemisia annua is an anti-fever herbal medicine first described in traditional Chinese medicine 1,000 years ago. Artemisinin,the extract of A. annua, and its derivatives(dihydroartemisinin(DHA), artemether, and artesunate) have been used for the treatment of malaria with substantial efficacy. Recently, DHA has also been tested for the treatment of lupus erythematosus,indicating that it may function to balance the immune response in immunocompromised individuals. In the present study, the regulatory effect of artemisinin on the murine immune system was systematically investigated in mice infected with two different protozoan parasites(Toxoplasma gondii and Plasmodium berghei). Our results revealed that the mouse spleen index significantly increased(spleen enlargement) in the healthy mice after DHA administration primarily due to the generation of an extra number of lymphocytes and CD8~+ T lymphocytes in both the spleen and circulation. DHA could increase the proportion of T helper cells and CD8~+ T cells, as well as decrease the number of splenic and circulatory B cells. Further, DHA could reduce the production of proinflammatory cytokines. Our study revealed that apart from their anti-parasitic activity, artemisinin and its derivatives can also actively modulate the immune system to directly benefit the host.     

Terpenoids from the seeds of Artemisia annua

Fourteen sesquiterpenes, three monoterpenes and one diterpene natural product have been isolated from the seeds of Artemisia annua. The possible biogenesis of some of these natural products are discussed by reference to recently reported experimental results for the autoxidation of dihydroartemisinic acid and other terpenoids from Artemisia annua.