绿木霉菌株T43对松烂皮病菌的抑制效果及其抑菌机理

通过对峙培养和非挥发性代谢产物对病原菌生长的抑制]作用,对引进绿木霉(Trichoderma virene)菌株T43及其抑菌活性物质对松树烂皮病菌(Cenangium ferruginosum)的抑制效果进行了研究;从菌株T43发酵液乙酸乙酯提取物对松树烂皮病菌的3种保护酶(SOD、CAT和POD)活力影响的角度,初步探讨了提取物对病原菌的抑菌机理.研究结果表明:对峙培养48 h后,菌株T43对松烂皮病菌相对抑制效果为1.00,72 h后菌株T43与病原菌形成明显的拮抗线,并抑制病原菌的生长.菌株T43乙酸乙酯提取物对病原菌株的室内生长抑制率为60.50％.菌株T43发酵液乙酸乙酯提取物对病原菌的3种保护酶具有明显的抑制作用.     

一株苏云金亚种的发酵产物对柳蚜3种保护酶活性的影响

为探究苏云金杆菌(Bacillus thuringiensis Bt)对蚜虫的生物活性影响及作用方式,以苏云金亚种(Bacillus thuringiensis subsp.thuringiensis)为目的菌株,以发酵产物为材料。利用5种不同浓度的菌液分别喷雾处理柳蚜,72h校正死亡率为29.74%～78.88%,其浓度与及死亡率呈正相关关系。苏云金杆菌亚种发酵液和发酵液滤液处理柳蚜后,12～24h柳蚜超氧化物歧化酶(SOD)比活力均被抑制,其抑制率分别为21.06%～38.11%和4.69%～36.79%,其中18h、24h对SOD比活力的抑制达显著水平(p<0.05)。发酵液和滤液处理组18h前对柳蚜过氧化氢酶(CAT)比活力的影响总体呈激活趋势,其中发酵液和滤液处理组分别在18h、12h对柳蚜CAT的激活率最高,其CAT比活力分别为对照的1.8890倍和2.9463倍。滤液处理组对柳蚜POD比活力总体呈激活作用,其激活率为14.78%～57.39%;发酵液处理组6h和18hPOD比活力显著高于对照,其他时间点POD比活力与对照无显著差异。     

青钱柳叶提取物对Caco-2细胞二糖酶活性的抑制作用

采用Caco-2细胞模型,分析了青钱柳Cyclocarya paliurus叶水提物、醇提后的水提物、95%醇提物、乙酸乙酯萃取物和正丁醇萃取物5种不同提取物对Caco-2细胞活力的影响及对Caco-2细胞上麦芽糖酶、蔗糖酶、乳糖酶3种二糖酶活性的抑制作用。结果表明,5种不同提取物中除95%醇提物对Caco-2细胞活力产生一定影响外,其余4种提取物对Caco-2细胞基本无毒性,在200 mg·L^-1浓度提取物范围内培养24～72 h后细胞仍能保持较高活力;对3种二糖酶活性抑制作用效果最好的为醇提后的水提物,尤其是对蔗糖酶和乳糖酶的抑制作用更为明显,最高的抑制率分别达到76.35%和60.13%,其次为乙酸乙酯萃取物,对乳糖酶的抑制率最高达54.41%。从青钱柳叶不同方法提取物的制备流程说明多糖类或中等极性的黄酮甙类有可能是青钱柳叶中抑制二糖酶活性的主要组成成分。     

杨树芽提取物体外抗肿瘤活性的研究

采用四甲基偶氮唑盐比色(MTT)法观察杨树芽提取物对MCF - 7、SMMC - 7721和A549 3种肿瘤细胞的体外生长抑制作用,并与市售蜂胶和杨树胶的抗肿瘤活性进行对比.结果表明:杨树芽提取物对3种肿瘤细胞作用48h的抑制率分别为97.1%、88.7%和60.5%,半数抑制质量浓度(IC_50)分别为58.4、83.9和196.3mg/L,均高于蜂胶和杨树胶.杨树芽提取物对体外培养的肿瘤细胞生长有明显的抑制作用,提示杨树芽提取物具有良好的抗肿瘤活性.     

杜仲叶提取物对人结直肠癌细胞HCT116及抗血管生成活性的影响

研究了杜仲叶提取物(EULE)对人结直肠癌细胞HCT116体外增殖和凋亡的影响以及其抗血管生成的活性。采用噻唑蓝(MTT)比色法测定了EULE及阳性对照药顺铂对HCT116和人脐静脉内皮细胞(HUVEC)体外增殖的影响;用流式细胞仪测定了EULE对HCT116细胞凋亡的影响。结果表明:EULE对HCT116细胞增殖具有一定的抑制作用,并表现出较好的量效关系,但抑制率稍低于顺铂;EULE对HCT116细胞的凋亡率的影响不显著;EULE对HUVEC细胞增殖的作用具有浓度依赖性,表现为低浓度促进增殖,高浓度抑制增殖。当EULE质量浓度分别为100、200、400和800 mg/L时,对HCT116细胞的抑制率分别为6.50%、10.52%、16.99%和56.49%,对HCT116细胞的凋亡率分别为0.00%、5.90%、7.00%和4.50%。EULE 800 mg/L时对HUVEC细胞的抑制率为41.59%,说明杜仲叶提取物只有达到一定浓度才具有抗新生血管生成的活性。     

核桃内生真菌G8菌株的分离鉴定及抑菌活性

从核桃根部分离得到一株真菌,编号为G8,对其进行生物学观察,初步鉴定为葡萄壳小圆孢菌,并对其进行了复侵染研究,确定其为核桃内生真菌.对此菌株进行液体发酵,研究其发酵液及其萃取物的抑菌活性.结果表明：（1）发酵液的提取物对28种植物病原真菌均有不同程度的抑制作用,各萃取相粗提物在40mg&#183;mL^-1时,乙酸乙酯萃取物的抑制效果最好,其次为正丁醇萃取物,萃余物最弱,但正丁醇萃取物对水稻纹枯、杨树溃疡、小麦雪腐病原菌的抑制率大于其它两相.（2）不同质量浓度的乙酸乙酯萃取物对11个病原真菌的活性实验结果表明：对苹果轮纹、苹果腐烂、小麦根腐、稻瘟4种病原菌的抑制率为100%,对苜蓿炭疽、小麦赤霉、番茄早疫、苹果炭疽和玉米小斑5种病原菌的抑制率表现出极显著差异,对玉米大斑和辣椒疫霉2种病原菌的抑制率无显著差异.（3）乙酸乙酯萃取物对5种细菌的抑菌试验表明,对蜡状芽孢杆菌和马铃薯芽孢杆菌2种细菌表现为抗药性,但不同质量浓度处理间差异不显著.     

瓦雷兹芽孢杆菌YH-20发酵液的抑菌特性初探

为掌握瓦雷兹芽孢杆菌YH-20发酵液的抑菌稳定性及其抑菌物质,采用生长速率法测定菌株YH-20发酵液对12株病原菌的抑制作用,以葡萄座腔菌为病原指示真菌测定发酵液在不同条件下抑菌稳定性,根癌农杆菌为病原指示细菌测定提取物质的抑菌活性;采用硫酸铵沉淀法和酸沉淀法提取发酵液中的蛋白类物质和脂肽类物质,并以特定培养基检测菌株产蛋白酶和纤维素酶的能力,以正丁醇、乙酸乙酯和石油醚对发酵液中的活性物质进行萃取。结果表明,菌株YH-20发酵液对供试的11种病原真菌和1株病原细菌均具有较好的抑制能力,该发酵液经普通光照和紫外光照处理后,发酵液的抑菌率保持稳定,经80℃处理120min后,抑菌率仍可达到49.24%,在pH6-9处理后,仍可保持较高的抑菌率。在4℃最少可储存30d而抑菌能力不发生改变。菌株YH-20可产生抑菌性蛋白和脂肽、蛋白酶、纤维素酶及嗜铁素。以极性较高的正丁醇和乙酸乙酯可萃取出发酵液中的抑菌活性物质。菌株YH-20对植物病原菌具有广谱抑菌能力和较好的抑菌稳定性,可产生多种抑菌物质。     

核桃叶中抗植物病毒活性物质提取条件的研究

以不同溶剂和方法对核桃叶进行了抗植物病毒物质的提取和活性测定.结果表明:提取物的抗病毒活性因提取溶剂和提取方法的不同而异.以体积分数 95 % 乙醇为提取溶剂,冷浸提取物在质量浓度 10 g/L 时,对烟草花叶病毒(TMV) 的抑制率最高,为 100 %.采用冷浸提取时,核桃叶中抗TMV活性物质提取的优化条件是:以体积分数 95 % 乙醇为提取溶剂,料液比1:10(g:mL),提取 18 h,得率 21.42 %.质量浓度 5 g/L 的提取物对TMV侵染心叶烟的抑制率为 100 %.该提取物的乙酸乙酯萃取物经多次层析分离和纯化得到黄酮类化合物A,质量浓度 0.5 g/L 的化合物A对TMV侵染心叶烟的抑制率达 85.55 %,说明化合物A是乙酸乙酯萃取物中的主要抗病毒活性成分之一.     

24种竹笋蛋白对肿瘤细胞增殖的抑制作用

目的:探讨不同种类竹笋蛋白对胃癌细胞SGC7901和结肠癌细胞HCT116增殖活性的影响。方法:采用植物活性蛋白提取试剂盒从24种不同种类竹笋中提取蛋白并进行BCA法定量,CCK-8细胞增殖活性检测方法检测不同竹笋蛋白对胃癌细胞SGC7901和结肠癌细胞HCT116增殖活性的影响;不同浓度竹笋蛋白作用胃癌细胞SGC7901和结肠癌细胞HCT116,72 h后采用显微镜拍照记录细胞生长情况。结果:24种竹笋蛋白均可不同程度地抑制胃癌细胞SGC7901和结肠癌细胞HCT116的增殖,同一种竹笋蛋白不同浓度的增殖抑制率不同,呈现出一定的浓度依赖关系;24种竹笋蛋白作用胃癌细胞SGC7901和结肠癌细胞HCT116,72 h后细胞生长受到明显的抑制。结论:竹笋蛋白对肿瘤细胞,如胃癌细胞SGC7901和结肠癌细胞HCT116均有显著的抑制作用,为肿瘤的发生发展的预防治疗提供了重要方法参考,具有一定的临床开发价值。     

异海松酸糠醛类酰腙化合物的合成及生物活性研究

以异海松酸为原料,采用先酰氯化后与水合肼反应的方法,制备得到异海松酸酰肼,然后再与不同取代基的糠醛发生反应,制备得到5种异海松酸糠醛类酰腙化合物:异海松酸基(糠醛)酰腙(4a)、异海松酸基(5-甲基糠醛)酰腙(4b)、异海松酸基(5-羟甲基糠醛)酰腙(4c)、异海松酸基(5-溴-2-糠醛)酰腙(4d)和异海松酸基(5-(4-溴苯基)糠醛)酰腙(4e),并采用FT-IR、1 H NMR、13 C NMR和MS对产物结构进行了确证。生物活性测定结果表明:目标化合物对肺炎链球菌、肺炎克雷伯氏菌、大肠杆菌、表皮葡萄球菌和金黄色葡萄球菌这5种菌种均具有一定的抑制活性,其中4a对肺炎克雷伯氏菌具有很好的抑菌活性,最低抑菌浓度为1.95 mg/L,化合物4c对肺炎链球菌具有非常好的抑菌活性,最低抑菌浓度仅为0.98 mg/L。当异海松酸糠醛类酰腙化合物的浓度为100μmol/L时,化合物4d对人体肝癌(Hep G2)、乳腺癌(MDA-MB-231)、前列腺癌(PC-3)和宫颈癌(Hela)这4种人体肿瘤细胞均具有较高的抑制率,其抑制率分别为75.17%、82.33%、78.52%和80.97%;化合物4e对人体肝癌(Hep G2)的抑制率高达93.68%,说明其对人体肝癌细胞具有很强的抑制活性。     

Antiproliferative activity of Saponaria vaccaria constituents and related compounds

Total methanolic extracts of Saponaria vaccaria seed derived from several varieties, as well as various purified components obtained through successive chromatographic separations of total extracts were evaluated for their growth inhibitory activity in WiDr (colon), MDA-MB-231 (breast), NCI-417 (lung) and PC-3 (prostate) human cancer cells as well as the non-tumorigenic fibroblast BJ (CRL-2522) cell line using MTT colorimetric assay. Purified bisdesmosidic saponins segetoside H and I were further examined using microscopy and apoptosis assays. Bisdesmosidic saponins exhibited dose-dependent growth inhibitory and selective apoptosis-inducing activity. Growth inhibitory effects were particularly strong in a breast (MDA-MB-231) and a prostate (PC-3) cancer cell line. Total extracts exhibited a different preference being most active against a colon cancer cell line (WiDr). In a comparison of varieties, all of the total seed extracts exhibited similar dose-dependent activities, but with some variation in potency. Monodesmosidic saponins vaccarosides A and B, phenolic vaccarin, and cyclopeptide segetalin A, co-occurring seed substituents, did not exhibit activity. The non-tumorigenic fibroblast cell line BJ (CRL 2522) was growth inhibited but did not undergo apoptosis when treated with bisdesmosidic saponins at low micromolar concentrations. Saponin-rich extracts from Kochia scoparia seed and Chenopodium quinoa were also evaluated alongside Saponaria saponins but did not exhibit activity. Closely related Quillaja saponins exhibited activity but were less potent.     

Antiproliferative effects of Ceratonia siliqua L. on mouse hepatocellular carcinoma cell line

Extracts from pods and leaves of carob (Ceratonia siliqua L.) were tested for their ability to inhibit cell proliferation of mouse hepatocellular carcinoma cell line (T1). The two extracts showed a marked alteration of T1 cell proliferation in a dose-related fashion reaching the maximal effect at 1 mg/ml. Moreover, we demonstrated that leaf and pod extracts were able to induce apoptosis in T1 cell lines after 24-h treatment mediating a direct activation of the caspase 3 pathway. HPLC analysis revealed the presence of gallic acid, (-) epigallocatechin-3-gallate and (-) epicatechin-3-gallate in pod and leaf extracts, compounds well known to exert antiproliferative effects. Their concentration reached 6.28 mg/g in carob leaves and 1.36 mg/g in carob pods extract. The discovery that carob pod and leaf extracts contained antiproliferative agents could be of practical importance in the development of functional foods and/or chemopreventive drugs.     

Isoflavone content and estrogenic activity of different batches of red clover (Trifolium pratense L.) extracts: An in vitro study in MCF-7 cells

The estrogenicity of different batches of red clover (Trifolium pratense L., Fabaceae; RCL) extracts and its relationship with the isoflavone content were assessed by measuring MCF-7 cell proliferation by flow cytometry and propidium iodide staining. RCL extracts were compared to estradiol (E2) and to the main RCL isoflavones biochanin A, daidzein, genistein and formononetin. Isoflavone content in the extracts was assayed by HPLC.E2 and isoflavones increased MCF-7 proliferation in a concentration-dependent fashion, with the following potency order: E2 >>> genistein > biochanin A = daidzein > formononetin. Extracts increased MCF-7 proliferation with different potencies, which in four out of five extracts correlated with the ratios 5,7-dihydroxyisoflavones/7-hydroxyisoflavones. The efficacy of all extracts increased with decreasing genistein contents. A solution containing the main isoflavones at the average concentration of RCL extracts increased MCF-7 proliferation with higher potency and steeper concentration–response curve. The effects of E2, of RCL extracts and of the isoflavone solution were inhibited by the estrogen receptor antagonist 4-hydroxytamoxifen.Flow cytometric analysis of MCF-7 proliferation is a suitable bioassay for the estrogenicity of RCL extracts, thus expanding the characterization of individual batches beyond assessment of chemical composition and contributing to improved standardization of quality and activity.     

Wound-healing plants from TCM: in vitro investigations on selected TCM plants and their influence on human dermal fibroblasts and keratinocytes

Wound-healing plants from Traditional Chinese Medicine and described for wound healing in the Pharmacopoeia of People's Republic of China (2005 ed.) were investigated by in vitro bioassay on human skin cells. Therefore water and EtOH–water extracts (6:4, v/v) from 12 plants were tested on human primary dermal fibroblasts (pNHDF) and human HaCaT keratinocyte cell line by quantification of cell viability (MTT assay) and cellular proliferation (BrdU incorporation ELISA). No functional activity was found for extracts from Achyranthis bidentatae rhizoma, Cimicifugae rhizoma, Corydalis rhizoma, Gardeniae fructus, Houttuyniae herba, Lonicerae japonicae caulis, Paeoniae rubrae radix and Rehmanniae radix. Extracts from Notoginseng radix et rhizoma, Angelicae sinensis radix and Lonicerae japonicae flos showed moderate activity, while extracts from Moutan cortex (the root bark of Paeonia suffruticosa Andr., Ranunculaceae) increased cell viability of HaCaT keratinocytes and pNHDF in a dose-dependent manner significantly. Bioassay-guided fractionation yielded paeonol 1, the flavan-3-ols catechin 2 and epicatechin-3-O-gallate 3, the dimeric proanthocyanidin epicatechin-(4β → 8)-catechin 4, a mixture of trigalloyl-glucoses 5 and 1,2,3,4,6-penta-O-galloyl-β-d-glucose (PGG) 6. The proanthocyanidin-containing fractions as well as PGG-containing fractions contributed substantially to the stimulating effects. Especially PGG-containing fractions enhanced cell viability and cellular proliferation of HaCaT keratinocytes at concentration of 100 nM.From these data we conclude that indication claims for TCM herbal materials must be carefully investigated in order to establish evidence-driven use of such plants. In case of Moutan cortex skin cell stimulating effects have clearly been proven. These effects can be related to the polyphenol fractions of condensed and hydrolysable tannins.     

Cytotoxic ellagitannins from Reaumuria vermiculata

Three ellagitannins and one disulfated flavonol were isolated from the aerial parts of Reaumuria vermiculata L. Besides that, 16 known compounds were characterized as well. The structures of all compounds were elucidated on the basis of spectroscopic data including 1D and 2D NMR and ESI HR-FTMS. The in vivo antioxidant activity using the oxygen radical absorbance capacity (ORAC) method, of the extract, its column fractions and two of the isolated ellagitannins was accomplished. In addition, a possible cytotoxicity of the extract and two of the new ellagitannins on HaCaT human keratinocytes and the activity of both compounds against the prostate cancer cell line (PC-3) were also assessed, whereby a potent cytotoxicity with IC(50) less than 1μg/ml was determined for both compounds. Besides, the extract exhibited a potential cytotoxic effect against four different solid tumor cell lines, namely liver (Huh-7), colorectal (HCT-116), breast (MCF-7) and prostate (PC-3). The IC(50)s were found to be substantially low (ranged from 1.3±0.15 to 2.4±0.22μg/ml) with relatively low resistance possibility reaching to 0% in the case of Huh-7 cell.     

不同类型油茶穗条提取液中生根抑制物的研究

为给油茶无性繁殖提供理论依据,以油茶新品种‘秋霞’为试验材料,对不同月份(4月、6月)采集的不同类型穗条(8年生嫁接大树、3年生组培苗、3年生家系苗、3年生扦插苗)提取液的酸度与酚类物质的含量进行了检测与分析;并以白菜种子为试材,就不同类型穗条提取液对其发芽的生物效应进行了鉴定试验。测定结果表明:不同月份采集的不同类型穗条提取液的酸度和酚类物质含量均不同。鉴定试验结果表明:白菜种子在不同月份采集的不同类型穗条提取液原液中的发芽率不存在显著差异,但白菜种子在嫁接大树穗条提取液中的发芽率最低,而在家系苗穗条提取液中的发芽率最高。研究结果表明:不同类型穗条提取液中均存在生根抑制物质,且均存在酸类和酚类物质;嫁接大树穗条中生根抑制物质的含量较高,而家系苗穗条中生根抑制物质的含量较低。     

Preliminary antiproliferative effects of some species of Terminalia, Combretum and Pteleopsis collected in Tanzania on some human cancer cell lines

Methanolic extracts (25 microug/ml) of species belonging to the genera of Combretum, Terminalia and Pteleopsis, collected during a field expedition in Tanzania in 1999, were screened for their antiproliferative and cytotoxic effects against three human cancer cell lines (HeLa, cervical carcinoma; T 24, bladder carcinoma; and MCF 7, breast carcinoma). A leaf extract of Combretum fragrans and a fruit extract of C. zeyheri gave the strongest antiproliferative and cytotoxic effects of all the twenty-four extracts screened in this investigation. In contrast to the highly powerful leaf extract of C. fragrans, the root extract of this species gave no cytotoxic effects against the investigated cancer cell lines at a concentration of 25 microg/ml. The other investigated species of Combretum and Terminalia differed greatly in their cytotoxic potential. Root extracts of Terminalia sambesiaca and T. sericea gave the strongest cytotoxic effects of the five species of Terminalia used in this study. Eight of the twenty-four investigated plant extracts showed pronounced cytotoxic effects (<30% proliferation compared to the control) against the T 24 bladder cancer cells, seven against the HeLa cells and four against the MCF 7 cells.     

Estrogenic activity of the dichloromethane extract from Pueraria mirifica

Pueraria mirifica and its extracts are widely used as the ingredient(s) in many rejuvenating products. Up to now, the extract of P. mirifica roots that has been used in most studies, is the alcoholic extract. In the present study, we investigated the estrogenic activity using uterotropic and MCF-7 cell proliferation models of the dichloromethane extract as well as the water extract which was obtained from partitioning the ethanolic extract. The results indicated that among the three extracts, i.e. the ethanolic extract (PM1), the water extract (PM2) and dichloromethane extract (PM3), PM3 exhibited the most potent estrogenic activity in both models, followed by PM1. The extracts produced uterotropic activity associated with the increase of water content while uterotropic activity of 17beta-estradiol was related to the increase of muscle mass. The two isoflavonoids, genistein and daidzein, were not the major active phytoestrogens involving the estrogenic activity of these extracts.     

The effects of ganoderma alcohols isolated from Ganoderma lucidum on the androgen receptor binding and the growth of LNCaP cells

The effects of ganoderma alcohols isolated from ethanol extracts of Ganoderma lucidum (Fr.) Krast (Ganodermataceae) on the androgen receptor binding and the growth of LNCaP cells have been investigated. Less than two hydroxyl groups in 17β-side chain are needed for binding to androgen receptor. In the case of the ganoderma alcohols with the same number of hydroxyl groups in 17β-side chain, the one which has C-3 carbonyl group showed better binding activity to androgen receptor than that which has C-3 hydroxyl group. The unsaturation in 17β-side chain is needed for the inhibition of the cell proliferation of androgen-induced LNCaP cells growth.