鬼臼类植物产鬼臼毒素内生真菌的筛选

从桃儿七(Sinopodoph yllum hexandrum)、南方山荷叶(Diphylleia sinensis)和川八角莲(Dysos-ma veitchii)中分离到92株内生真菌,其发酵提取物经TLC、HPLC,结果表明,6株内生真菌能产鬼臼毒素,占总分离菌株数的6．5％。桃儿七、南方山荷叶和川八角莲内生真菌中产鬼臼毒素菌株占各供测菌株的7％,3％和10％。     

川八角莲内生真菌产鬼臼毒素类似物的初步研究

目的：从川八角莲中分离到产抗癌药物鬼臼毒素类似物的内生真菌。方法：采用薄层层析法(TLC)和高效液相色谱法(HPLC)对内生真菌的发酵产物进行分析检测。结果：分离到16株内生真菌,其中一株真菌(2BNO1)产鬼臼毒素类似物,经鉴定该真菌为牵连青霉(Penicillium implication)。结论：川八角莲个别内生真菌能产抗癌药物鬼臼毒素类似物。     

川八角莲的化学成分

川八角莲〔Dysosma veitchii (Hemsl. el Wils.) Fu (Podophyllum V.)〕为鬼臼科植物,我国主要分布在四川、云南、贵州。其根之乙醇提取物显示强的抗癌活性。我们从乙醇提取物主氯仿溶解部份分得三个木脂素:去氧鬼臼毒素(deoxypodophyllotoxin(1),β—足叶草脂素(β-peltatin) (2)和鬼臼毒素(podophyllotoxin) (3);从氯仿不溶部份分到山萘黄素3—β—D—吡喃葡萄糖苷(kaempfcrol 3—β—D—glucopyranoside)。另外,β—谷甾醇、葡萄糖、蔗糖亦从该植物根中分到。     

峨眉山的八角莲属植物

小檗科八角莲属(Dysosma)为我国特有的多型属植物类群。全属8种1变种,其中包括1987年以来在峨眉山发现的新种峨眉八角莲(D.emeiensis)和川八角莲变种即长瓣八角莲(D.veitchii var.longipetalis)。峨眉山计有本属植物5种1变种,尤其是集中了川八角莲(D.veltchii.)贵州八角莲(D.majorensis)     

西藏八角莲与桃儿七根及根茎SSH文库的建立

以西藏八角莲(Dysosma tsayuensis Ying)和桃儿七[Sinopodophyllum hexandrum(Royle)Ying]为材料,构建其根及根茎的SSH文库,从中筛选鬼臼类植物属种间与鬼臼毒素生物合成相关的差异表达基因。从文库中随机挑取201个阳性克隆测序后得到183条ESTs。去除载体序列和冗余序列,聚类拼接得到17个西藏八角莲的unique ESTs。经BLAST同源比较和功能查寻,有功能注释的unique ESTs共12个,占70.6%,所编码的蛋白涉及光合作用、合成代谢、转录调控等功能;无功能注释和匹配结果的共5个,占29.4%。该研究成功构建了西藏八角莲和桃儿七SSH文库,为进一步揭示鬼臼毒素生物合成途径及其调控机制奠定了基础。     

鬼臼毒素及其衍生物生物合成研究进展

鬼臼毒素(Podophyllotoxin,PTOX)是来源于中药八角莲、山荷叶和桃儿七等鬼臼属植物的芳基四氢萘类木脂素.其化学半合成衍生物依托泊苷和替尼泊苷被用于多种癌症的临床治疗.作为天然产物来源新药创制的典型代表,鬼臼毒素目前依赖天然提取,供求矛盾日渐突出.生物合成具有不受资源限制、反应条件友好等优势,是鬼臼毒素及...     

川八角莲繁殖生态学初步研究

对川八角莲(Dysosma veitchii)3个种群进行的繁殖生态学研究结果表明:川八角莲主要分布在常绿阔叶林下,个体在种群内呈片丛状分布或为散生。川八角莲只进行有性生殖,由种子萌发至性成熟约需要5～6年。性成熟植株的花芽在开放前一年8月上旬开始形成,至11月上旬营养生长结束时花各部分已分化完全。川八角莲为自花受粉植物,限制结实率与结籽率的主要因素是花的构造不利于授粉以及营养竞争。果实与种子的近距离散布主要通过重力和蚂蚁,远距离散布主要通过鸟和啮齿类食用果实来进行。在自然条件下,川八角莲不进行营养繁殖,但在人工条件下,可用根来进行营养繁殖。     

两种八角莲内生真菌的生物活性研究

测定云南八角莲Dysosma aurantiocaulis和川八角莲Dysosma veitchii内生真菌的生物活性,为进一步从内生真菌中寻找活性成分提供指导,也为认识内生真菌相互作用提供有益的参考。从两种八角莲中分离出内生真菌187株,用滤纸片法测抗菌活性,用卤虫模型测抗卤虫活性得知大部分菌株发酵液都有活性,高活性菌株主要集中在镰孢霉属Fusarium、青霉属Penicillium、木霉属Trichoderma、丛梗孢属Monilia、毛壳菌属Chaetomium、无孢类群Mycellia sterilia、小齿梗霉属Rhinotrichum、无梗孢属Trichocladium和胶帚霉属Gliocladium。云南八角莲和川八角莲内生真菌是活性成分筛选的重要来源,并且通过活性分析发现:发酵液的活性强于菌丝体;云南八角莲的抗细菌活性菌株多于川八角莲。     

广西八角莲与八角莲光响应特性的比较研究

为了解濒危植物广西八角莲对环境光强的适应性,该研究以广西八角莲同属渐危种八角莲为对照,采用Li-6400便携式光合测定系统对两种植物的光合光响应特性进行了比较研究,进而探讨广西八角莲的濒危机制。结果表明:广西八角莲与的八角莲的光饱和点分别为440和530μmol·m~(-2)·s~(-1),光补偿点为13.25和13.10μmol·m~(-2)·s~(-1),最大净光合速率为3.62和6.81μmol·m~(-2)·s~(-1),表观量子效率为0.065和0.042μmol·μmol~(-1),两种八角莲均具阴生草本植物的光合特性,但其光补偿点与饱和点均高于一般阴生草本,10%~30%阴蔽度的林下生境有利于两种八角莲的生长;两种植物相比较,广西八角莲的光合能力较弱,光饱和点较低,但其弱光下的量子效率较高。大部分光强下,八角莲的净光合速率、气孔导度和蒸腾速率均高于广西八角莲,但广西八角莲的瞬时水分利用效率却高于八角莲,表明广西八角莲的光合策略比较保守,以较低的光合积累为代价来维持较高的水分利用效率,以保持体内水分平衡。     

甘肃不同居群桃儿七[Sinopodophyllum hexandrum(Royle) Ying]鬼臼毒素含量与环境因子的相关性

为探讨桃儿七(Sinopodophyllum hexandrum)中鬼臼毒素含量与环境因子的相关性,采集甘肃省境内6个不同居群桃儿七的地下部分和根际土壤样品,测定鬼臼毒素含量和根际土壤中水分、pH值、有机质、总氮、速效磷、速效钾、有效铁、有效锰、有效铜、有效镁、总铁、总锰、总铜、总镁等14个因子。采用逐步回归分析和通径分析方法对土壤因子、海拔和气候因子(年均温、年均降雨量、年均日照时数、年均最高温(2010—2014年最高温的平均值)、年均最低温(2010—2014年最低温的平均值))与桃儿七地下部分的鬼臼毒素含量进行了相关性分析。结果表明:不同居群间桃儿七鬼臼毒素含量呈现极显著差异,居群内鬼臼毒素含量相对一致,基本呈正态分布。逐步回归结果显示,年均温、年均日照时数和土壤总氮含量是影响桃儿七鬼臼毒素含量的3个主导环境因子,且均与鬼臼毒素含量呈显著负相关。进一步通径分析表明,3个主导因子对鬼臼毒素含量的影响程度依次为:年均温年均日照时数总氮含量。研究结果表明,在桃儿七人工栽培中可通过适当低温、缩短日照时间、减施氮肥等措施来调控桃儿七鬼臼毒素含量。     

Low temperature enhanced the podophyllotoxin accumulation vis-a-vis its biosynthetic pathway gene(s) expression in Dysosma versipellis (Hance) M. Cheng – A pharmaceutically important medicinal plant

Dysosma versipellis (Hance) M. Chang is an endemic and endangered Chinese medicinal herb. Podophyllotoxin, a lead compound for the semi-synthesis of clinically useful anticancer compounds, is the main active aryltetralin lignan accumulated in D. versipellis. In this study, morphology and physiology traits in response to podophyllotoxin accumulation and its biosynthetic pathway genes expression were investigated in D. versipellis plants grown at low temperature (4∼6 °C) and greenhouse (25∼30 °C) temperature. Our study found that low temperature significantly increased plant biomass (2.7-fold), chlorophyll, and carotenoid content compared to greenhouse temperature. The mRNA expression analysis of podophyllotoxin pathway genes secoisolariciresinol dehydrogenase and pinoresionol lariciresinol reductase expression levels were also up-regulated by low temperature. In addition, plants grown at low temperature markedly increased podophyllotoxin accumulation to 3.49-fold in rhizome of D. versipellis than the greenhouse temperature. These results suggest that plants at low temperature enhance the accumulation of podophyllotoxin in rhizome of D. versipellis by over-expressing podophyllotoxin pathway genes. This study finding could be useful in the producing pharmaceutically important podophyllotoxin without overexploiting this endangered species.     

小檗科鬼臼亚科植物的核型研究

本文首次报道了中华山荷叶与川八角莲的核型,分别为Ｋ（２ｎ）＝１２＝８ｍ（４ＳＡＴ）＋２ｓｔ＋２ｔ及Ｋ（２ｎ）＝１２＝４ｍ（２ＳＡＴ）十４ｓｍ＋２ｓｔ（２ＳＡＴ）＋２ｔ,核型类型均为ＺＡ型。本文报道的桃儿七及八角莲的核型与前人的结果有一定差异,前者为：Ｋ（２ｎ）＝１２＝６ｍ（４ＳＡＴ）＋２ｓｍ＋２ｓｔ＋２ｔ,２Ｂ型,后者为Ｋ（２ｎ）＝１２＝８ｍ（２ＳＡＴ）＋２ｓｔ（２ＳＡＴ）＋２ｔ,为２Ａ型。本文分析了小檗科鬼臼亚科４个属共７种植物的核型,结果是该类植物的核型极为相似,染色体数目均为２ｎ＝１２,由８条ｍ或ｓｍ,２条ｓｔ以及２条ｔ染色体组成。核型的相似性反映了这类植物的亲缘关系,这４个属的植物是一个自然类群。但随着系统发育,核型的不对称性有所增加,其中以山荷叶属最为对称,八角莲属居中,桃儿七属与足叶草属最不对称。笔者认为,核型上的高度相似是该类植物在系统发育上不发达,属内种类稀少,通常为寡种属或单种属的重要原因。     

Interest of lignans in prevention and treatment of cancers

Lignans are diphenolic compounds widely distributed in the plant kingdom. They are mainly localised in lignified tissues, seeds and roots. These molecules are involved in plant defence mechanisms, but are also interesting for human health. Flax lignans belonging to the phytoestrogens are metabolised after ingestion into enterolignans that may offer a protection against the onset and development of hormono-dependant cancers. In vitro studies based on mammalian cellular models tend to confirm their beneficial effects observed during epidemiological studies and give us insights about their mechanisms of action. The most studied lignan, podophyllotoxin, and its semi-synthetic derivatives (etoposide, teniposide, etoposide phosphate), are particularly interesting at a curative level due to their cytotoxic properties. These semi-synthetic derivatives are used in chemotherapy of lung cancer for example. However, the extensive use of these anticancer drugs will lead to the problem of podophyllotoxin supply. This molecule is currently extracted from the rhizomes and roots of an Indian species Podophyllum hexandrum which has subsequently become endangered. Strategies are investigated to obtain economically viable alternative sources of Podophyllotoxin from plants and in vitro cultures of several species. Among them, north american Podophyllum peltatum, Linum wild species, Hyptis, Anthriscus, Juniperus or Dysosma species which accumulate Podophyllotoxin or closely related derivatives, are good candidates. double dagger.     

鬼臼类植物内生真菌的分离及其抗癌活性研究

通过对三属四种鬼臼类植物地下茎内生真菌的分离,发现鬼臼类植物地下茎中内生真菌的物种类型极其丰富,主要分布在地下茎的表皮层和维管组织中,来源于外界环境.通过对包括鬼臼类植物在内的7种植物内生真菌的抗癌活性测定,发现内生真菌的抗癌活性与宿主有密切有关系,鬼臼类植物地下茎内生真菌含有较高比例的抗癌活性菌株.宿主种类、地理位置都会影响内生真菌的分布,进而影响活性菌株出现的频率.通过对所有鬼臼类植物地下茎内生真菌次生代谢产物的深入分析,并没有发现产生鬼臼毒素的内生真菌,鬼臼类植物地下茎内生真菌的抗癌活性成分是独立产生的.     

Antitumor agents. Part 212. Bucidarasins A-C, three new cytotoxic clerodane diterpenes from Bucida buceras

As part of a study on antitumor agents from rainforest plants, four new clerodane diterpenes, bucidarasins A--D (1-4), were isolated from Bucida buceras. Their structures were elucidated from detailed 2D NMR analyses. Compounds 1-3 showed potent cytotoxicity against human tumor cell lines with IC(50) values of 0.5-1.9 microM. The potency was retained in drug resistant lines.     

Antiproliferation and cell apoptosis inducing bioactivities of constituents from Dysosma versipellis in PC3 and Bcap-37 cell lines

Background

Recently, interest in phytochemicals from traditional Chinese medicinal herbs with the capability to inhibit cancer cells growth and proliferation has been growing rapidly due to their nontoxic nature. Dysosma versipellis as Bereridaceae plants is an endemic species in China, which has been proved to be an important Chinese herbal medicine because of its biological activity. However, systematic and comprehensive studies on the phytochemicals from Dysosma versipellis and their bioactivity are limited.

Results

Fifteen compounds were isolated and characterized from the roots of Dysosma versipellis, among which six compounds were isolated from this plant for the first time. The inhibitory activities of these compounds were investigated on tumor cells PC3, Bcap-37 and BGC-823 in vitro by MTT method, and the results showed that podophyllotoxone (PTO) and 4'-demethyldeoxypodophyllotoxin (DDPT) had potent inhibitory activities against the growth of human carcinoma cell lines. Subsequent fluorescence staining and flow cytometry analysis indicated that these two compounds could induce apoptosis in PC3 and Bcap-37 cells, and the apoptosis ratios reached the peak (12.0% and 14.1%) after 72 h of treatment at 20 μM, respectively.

Conclusions

This study suggests that most of the compounds from the roots of D. versipellis could inhibit the growth of human carcinoma cells. In addition, PTO and DDPT could induce apoptosis of tumor cells.