Anti-fungal effects of phenolic amides isolated from the root bark of Lycium chinense

Four phenolic amides, dihydro-N-caffeoyltyramine (1), trans-N-feruloyloctopamine (2), trans-N -caffeoyltyramine (3), and cis-N-caffeoyltyramine (4), were isolated from an ethyl acetate extract of the root bark of Lycium chinense Miller. All had an anti-fungal effect; compounds 1-3 were potent at 5-10 microg ml(-1) and were without hemolytic activity against human erythrocyte cells. Compound 4 was active at 40 microg ml(-1). All four compounds impeded the dimorphic transition of pathogen, Candida albicans.     

Caesalpinia palmeri: First Report on the Phenolic Compounds Profile,Antioxidant and Cytotoxicity Effect

This study aimed to determine the phenolic compounds profile, antioxidant potential and cytotoxicity of extracts and fractions of Caesalpinia palmeri. Methanolic extracts were generated from C. palmeri berries, stems and flowers. The latter was subjected to liquid-liquid partition, obtaining hexane, ethyl acetate and residues fractions. Results showed that the flower extract and ethyl acetate fraction had a larger concentration of phenolic compounds (148.9 and 307.9 mg GAE/g, respectively), being ellagic acid (6233.57 and 19550.08 μg/g, respectively), quercetin-3-β-glycoside (3023.85 and 8952.55 μg/g, respectively) and gallic acid (2212.98 and 8422.34 μg/g, respectively) the most abundant compounds. Flower extract and ethyl acetate fraction also presented the highest antioxidant capacity on all tested methods (DPPH, ABTS, ORAC and FRAP) and low cytotoxicity against ARPE-19 cells (IC₅₀ >170 μg/mL). C. palmeri possessed high antioxidant potential, associated with the presence of phenolic compounds and low cytotoxicity, suggesting that they could represent an option to counter oxidative stress.     

朱红栓菌提取物抗氧化、抗肿瘤活性评价及相关化学成分分析

采用石油醚、乙酸乙酯、乙醇浸提朱红栓菌 Trametes cinnabarina 子实体干粉,得到不同极性提取物;采用清除DPPH 自由基、羟自由基、超氧阴离子自由基能力,测定提取物的体外抗氧化活性;MTT法检测提取物对人肝癌细胞株HepG2细胞增殖的抑制作用。结果表明,朱红栓菌石油醚、乙酸乙酯、乙醇提取物均具有一定的抗氧化、抗肿瘤活性;各提取物在浓度为4-5mg/mL时,对DPPH自由基、羟自由基和超氧阴离子自由基清除能力大小依次为乙酸乙酯提取物>乙醇提取物>石油醚提取物;乙酸乙酯提取物对3种自由基的最高清除率分别为60.23%、74.49%、63.84%。各提取物对人肝癌细胞株HepG2细胞增殖抑制作用大小依次为乙酸乙酯提取物>乙醇提取物>石油醚提取物;乙酸乙酯提取物的抑制率最高达55.93%。采用硅胶和凝胶等柱色谱方法结合核磁、波谱和质谱等技术对乙酸乙酯提取物的化学组分进行分析,共分离纯化出11种化合物,分别鉴定为：麦角甾醇（1）,邻苯二甲酸二丁酯（2）,对羟基苯甲酸甲酯（3）,麦角甾-7,22,二烯-3-酮（4）,1-[(12E,16E)-12,16-二十碳二烯酰基]-2-[(E,E)-7,11-十八碳二烯酰基]-3-硬脂酰基甘油（5）,cinnabarin（6）,过氧麦角甾醇（7）,尿嘧啶（8）,甘露醇（9）,腺嘌呤核苷（10）,豆甾-7,22-二烯-3β,5α,6β-三醇（11）。除化合物6外均为首次从朱红栓菌子实体中分离得到。研究结果为开发利用朱红栓菌子实体提供了科学依据。     

Cytotoxic,hypoglycemic activity and phytochemical analysis of Rubus imperialis (Rosaceae)

Rubus imperialis, Artemia salina, 3-O-methylellagic-4'-O-alpha-rhamnose Acid Screening of different extracts, fractions and compounds from Rubus imperialis Chum. Schl. (Rosaceae) has been conduced using the brine shrimp microwell cytotoxicity assay. Three parts of the plant (methanolic extract from leaves, roots and stems), three fractions from roots (hexane, ethyl acetate and butanol) and three isolated compounds (niga-ichigoside F1, 23-hydroxytormentic acid, ellagic acid derivative) were tested. The most promising material (LC50 <1000 microg/ml) were the methanolic extract and ethyl acetate fraction from roots. However, there was little correlation observed in the degree of toxicities observed between the isolated compounds. On the other hand, the cytotoxicity and in vivo assays confirmed the hypoglycemic activity of methanolic extract and validated the Brazilian popular use of R. imperialis as an antidiabetic agent.     

Androgenic action of Tinospora cordifolia ethanolic extract in prostate cancer cell line LNCaP

Aim of the studyRecently, Tinospora cordifolia (TC) was shown to affect prostate growth in rats. It is not known whether this is a direct effect of TC or whether it is induced by altered hormone release. To investigate the actions of TC on the prostate, human LNCaP cells were exposed to an ethanolic extract of TC.Materials and MethodsLNCaP cells were incubated with the test substances for 48 h. Proliferation was measured by MTT test and prostate-specific antigen (PSA) secretion was determined with ELISA.ResultsTC showed a dose-dependent stimulation of proliferation of LNCaP cells. Co-incubation with the anti-androgen flutamide (FLU) reversed the TC-induced stimulation of PSA secretion.ConclusionsThe reference compound dihydrotestosterone (DHT) caused a significant increase of growth of LNCaP cells. Similarly, TC stimulated proliferation of these prostate cells. The anti-androgen FLU reversed the increase of PSA release caused by either DHT or TC. Thus, we suggest that TC may contain androgenic compounds, which appear to act via androgen receptor (AR).     

苦瓜叶提取物对美洲斑潜蝇取食和产卵行为的抑制作用

美洲斑潜蝇是危害蔬菜、观赏植物的重大害虫之一.苦瓜叶乙醇提取物(浓度为2000～4000 μg·ml^-1)对美洲斑潜蝇成虫的取食和产卵都具有较强的抑制作用.用环己烷、乙酸乙酯、正丁醇和水依次对乙醇提取物进行萃取,并测试了4种萃取物对美洲斑潜蝇成虫取食和产卵的抑制作用.结果表明: 环己烷、乙酸乙酯、正丁醇和水萃取物在浓度为1000 μg·ml^-1时,处理后2 d对美洲斑潜蝇成虫的拒食率分别是11.08%、34.89%、22.99%和 0,产卵抑制率分别是0、30.91%、6.45%和 0.其中,乙酸乙酯萃取物的活性最强,当其浓度为4000 μg·ml^-1时,处理后2 d对美洲斑潜蝇成虫的拒食率和产卵忌避率分别为70.95％ 和69.49％.乙酸乙酯萃取物经硅胶柱层析分离得到(19S,23E)-5β,19-环氧-19-甲氧葫芦素-6,23-二烯-3β,25-二醇 (化合物1)、(19R,23E)-5β,19-环氧-19-甲氧葫芦素-6,23-二烯-3β,25-二醇(化合物2) 和3β,7β,25-三羟基葫芦素-5,23-二烯-19-醛缩-3-O-β-D-吡喃葡糖苷（化合物3）,3种化合物在供试的浓度（100～400 μg·ml^-1）条件下对美洲斑潜蝇的取食和产卵行为都有明显的抑制作用.在400μg·ml^-1浓度时,化合物1、化合物2和化合物3对美洲斑潜蝇成虫的拒食率分别是66.89%、53.53%和78.02%,产卵抑制率分别是76.32%、58.36%和78.36%.     

甘蓝型油菜花粉离体培养

通过对甘蓝型油菜花粉发育阶段和活力的检测确定花粉发育的时期,分离出单核晚期花粉进行离体培养.结果表明,(1)筛选出适合油菜小孢子花粉离体培养的液体培养基为T_1+怀特维生素(White's vitamins)+2%椰子汁+0.5 mol/L麦芽糖,在此培养基上花粉的成熟率可达25.1%,萌发率达6.3%.(2)筛选出适合成熟花粉离体萌发液体培养基为0.6 mol/L麦芽糖+1.6 mmol/L硼酸+2.9 mmol/L硝酸钙+29.6 μmol/L VB_1,在此培养基上,自然成熟花粉的萌发率可达75.2%.将离体培养成熟的花粉培养在萌发培养基,萌发的花粉占成熟花粉的66.3%.     

三叶蔓荆子提取物对小菜蛾的拒食活性

室内测定了三叶蔓荆子(Vitex trifolia(L.))叶片提取物对小菜蛾Plutella xylostella(L.)3龄幼虫的拒食活性。非选择性试验结果表明,三叶蔓荆子的不同溶剂(乙酸乙酯、石油醚、氯仿和乙醇)提取物对小菜蛾均有一定的拒食作用。4种提取物对小菜蛾的毒力顺序为:乙酸乙酯提取物>石油醚提取物>氯仿提取物>乙醇提取物。选择性试验表明,乙酸乙酯提取物对小菜蛾24、48h的拒食中浓度(AFC50)分别为2 520、3240mg.L-1。室内盆栽试验结果表明,施用乙酸乙酯提取物(10000mg.L-1)3d后甘蓝(Brassica oleracea var.capitata)植株上3龄小菜蛾的虫口减退率可达72.76%。     

蚁巢膝束霉次级代谢产物及生物活性

菌株L342分离自中国四川省的废弃白蚁巢,rDNA序列分析表明L342为蚁巢膝束霉Geniculisynnema termiticola。抗氧化活性评价结果显示其PDB发酵产物的乙酸乙酯提取物具有很强的还原能力和中等强度的DPPH清除能力。采用硅胶柱色谱、ODS柱色谱、Sephadex LH-20柱色谱、HPLC等分离方法,从G. termiticola的PDB发酵粗提物中分离鉴定了6个化合物：3,4-Dihydro-6,8-dihydroxy-3-methylisocoumarin (1), regiolone (2), (3S, 4S)-3,4,6,8-tetradydroxy-3,4-Dihydronaphthalen-1(2H)-one (3), ergosta-4,6,8(14),22-tetraen-3-one (4), cerebroside A (5), and cerebroside C (6)。其中化合物4-6具有中等强度的铁离子还原能力,化合物4还具有一定的HeLa细胞毒活性（IC50=98.3μmol/L）。G. termiticola发酵产物具有产生有生物活性次级代谢产物的潜力,有望开发成为一种新型的功能性食品。     

EGFR signaling pathway negatively regulates PSA expression and secretion via the PI3K-Akt pathway in LNCaP prostate cancer cells

Epidermal growth factor (EGF) and its receptor (EGFR) are involved in hormone-refractory growth and poor prognosis of a subgroup of human prostate cancer. In this communication, we investigated the regulation of PSA by the EGFR signaling pathway using LNCaP C-81 prostate cancer cells. Administration of EGF stimulated the growth of LNCaP C-81 cells, however, PSA expression and secretion were suppressed. An EGFR inhibitor, AG1478, abrogated the PSA suppression effect by EGF, in concurrence with the suppression of tyro-phosphorylation levels of EGFR. Interestingly, the AR level was also decreased in EGF-treated LNCaP C-81 cells. Moreover, LY294002, but not PD98059, inhibited the PSA and AR suppression effect by EGF in concurrence with the suppression of phosphorylation levels of Akt. In conclusion, our results strongly suggest the existence of a novel androgen-independent PSA regulatory mechanism, i.e., the EGFR signaling pathway negatively regulates PSA expression which may be induced by the alteration of AR expression via the PI3K-Akt pathway in LNCaP C-81 cells.     

Secretion of endogenous kallikreins 2 and 3 by androgen receptor-transfected PC-3 prostate cancer cells

Androgen independent PC-3 cells lack androgen receptor (AR) expression and do not produce kallikrein 2 (hK2) or 3 (prostate-specific antigen, PSA). In this paper, we examined the ability of androgens to stimulate PSA and hK2 production in AR transfected PC-3 cells (PC-3(AR)) and compared this to LNCaP cells. PSA and hK2 were measured in the culture medium and cell lysates using an ELISA-based immunofluorometric assay. Only androgens were able to induce PSA and hK2 secretion in PC-3(AR) cells in a dose- and time-dependent manner depending on the level of AR present. The level of androgen-induced PSA and hK2 secretion in PC-3(AR) cells was approximately 1.5 and 0.9% that induced in LNCaP cells, respectively. Insulin-like growth factor-I (IGF-I), which has been shown to activate AR in the absence of ligand, did not activate PSA secretion in the absence of androgen, but further increased the dihydrotestosterone-induced PSA secretion in PC-3(AR) cells. The lack of PSA and hK2 production in parental PC-3 cells is thus a result of their lack of AR expression. PSA and/or hK2 production in PC-3(AR) cells can thus serve as an endogenous reporter system to investigate AR action or to screen putative endocrine disrupters.     

Evaluation of cytotoxic compounds from calligonum comosum L. growing in Egypt

Calligonum comosum (Polygonaceae), an Egyptian desert plant, was extracted and fractionated using petroleum ether, methylene chloride, and ethyl acetate. The total methanolic extract and other fractions were tested for their anticancer activity using Ehrlich ascites, brine shrimp and antioxidant assays. Ethyl acetate fraction proved to be the most active in all assays. Eight compounds were isolated, purified, and identified from this fraction as (+)-catechin (1), dehydrodicatechin A (2), kaempferol-3-O-rhamnopyranoside (3), quercitrin (quercetin-3-O-rhamnopyranoside) (4), beta-sitosterol-3-O-glucoside (5), isoquercitrin (quercetin-3-O-glucopyranoside) (6), kaempferol-3-O-glucuronide (7), and mequilianin (quercetin-3-O-glucuronide) (8). All isolated compounds were tested for their cytotoxicity and antioxidant activity. Compound 2 showed the best cytotoxic and antioxidant activity.     

银合欢豆荚中的酚类化学成分

为探讨银合欢(Leucaena leucocephala)的化学成分,从银合欢豆荚乙醇提取物中分离得到7个酚类化合物,经过波谱分析,鉴定为原儿茶酸乙酯(1)、丁香酸(2)、3-羟基-1-(3-甲氧基-4-羟基苯基)丙烷-1-酮(3)、咖啡酸甲酯(4)、(Z)-对香豆醛(5)、3-甲氧基-4-羟苯丙烷-7,8,9-三醇(6)、愈创木基甘油-8-O-4′-芥子醇醚(7)。所有化合物均为首次从该植物中分离得到。化合物1对大肠杆菌和鼠伤沙门氏菌有一定的抑制活性。     

毛头鬼伞子实体化学成分及抗糖尿病活性初探

从毛头鬼伞子实体中萃取得到乙醇、乙酸乙酯、石油醚3种有机提取物,采用α-葡萄糖苷酶活性抑制实验对3种有机提取物的抗糖尿病活性进行评价,结果显示,乙酸乙酯提取物对α-葡萄糖苷酶有较强的抑制活性。采用柱层析技术从乙酸乙酯提取物中分离纯化出10种化合物,经核磁等方法鉴定为：（1）顺,顺-9,12-十八(碳)二烯酸;（2）顺式-9-十八烯酸;（3）(22E,24R)-麦角甾烷-5,7,22-三烯-3β醇;（4）3β-5α-6α-22E-麦角甾-7,22-双烯-3,5,6-三醇-6-亚油酸酯;（5）3β-5α-6α-22E-麦角甾-7,22-双烯-3,5,6-三醇-6-油酸酯;（6）邻苯二甲酸二异丁酯;（7）对羟基苯乙醇;（8）4-羟基苯乙基乙酸酯;（9）3-(4-羟基-3-甲氧苯基)败脂酸;（10）N-反式-3,4亚甲二氧基肉桂酰基-3-甲氧基酪胺。对分离化合物的α-葡萄糖苷酶活性抑制实验结果显示,N-反式-3,4亚甲二氧基肉桂酰基-3-甲氧基酪胺对α-葡萄糖苷酶具有较强的抑制活性,其IC₅₀值为4.17mg/mL。     

瑞香狼毒根的抑菌活性研究（Ⅰ）

以苹果干腐病菌、小麦赤霉病菌、番茄早疫病菌、南瓜枯萎病菌、玉米大斑病菌、烟草赤星病菌和辣椒疫霉病菌为供试菌,采用生物活性跟踪法、生长速率测定法和系统溶剂提取法对瑞香狼毒根中的杀菌活性物质进行了筛选和分离。结果表明,杀菌活性物质主要集中在乙酸乙酯提取物和甲醇粗提物中。从乙酸乙酯提取物中分离出两种杀菌活性物质AF1-4和AF1-5,同时还发现瑞香狼毒根中还存在增菌物质。     

模拟氮沉降对青藏高原地区油菜幼苗期生长和生理特性的影响

以甘蓝型油菜(青杂303)为研究材料,通过研究分析油菜幼苗期的生长和生理特性,旨在探讨油菜幼苗对青藏高原地区未来大气氮沉降的响应机制。实验以NH4NO3氮肥模拟大气氮沉降的NH4+、NO3-输入并分别喷施0、2.5、5.0 g/m2.a模拟青藏高原地区大气氮沉降增加的0倍、2倍、4倍背景。结果表明:施氮处理对油菜幼苗生长具有促进作用,油菜幼苗的地上部分干重、叶面积和株高等形态指标都发生了显著变化;随着施氮量增加油菜幼苗叶片的抗氧化酶活性增强,氮肥输入量为5.0 g/m2.a的处理组中,其幼苗叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性与对照的相比均具有极显著差异。因此,反映出青藏高原地区生态系统中植物生长受到有效氮的限制,在外界有效氮输入下,对植物生长发育具有促进作用。     

Antimutagenic activity of flavonoids from Chrysanthemum morifolium

A methanol extract from the flower heads of Chrysanthemum morifolium showed a suppressive effect on umu gene expression of the SOS response in Salmonella typhimurium TA1535/pSK1002 against the mutagen 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide). The methanol extract was re-extracted with hexane, chloroform, ethyl acetate, butanol, and water. The ethyl acetate fraction showed a suppressive effect. Suppressive compounds in the ethyl acetate fraction were isolated by silica gel column chromatography and identified as the flavonoids acacetin (1), apigenin (2), luteolin (3), and quercetin (4) by EI-MS, IR, and (1)H and 13C NMR spectroscopy. Compounds 1-4 suppressed the furylfuramide-induced SOS response in the umu test. Compounds 1-4 suppressed 60.2, 75.7, 90.0, and 66.6% of the SOS-inducing activity at a concentration of 0.70 micromol/ml. The ID50 (50% inhibitory dose) values of 1-4 were 0.62, 0.55, 0.44, and 0.59 micromol/ml. These compounds had the suppressive effects on umu gene expression of the SOS response against other mutagens, 4-nitroquinolin 1-oxide (4NQO) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), which do not require liver-metabolizing enzymes. These compounds also showed the suppression of SOS-inducing activity against the other mutagens aflatoxin B1 (AfB1) and 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), which require liver-metabolizing enzymes, and UV irradiation. In addition to the antimutagenic activities of these compounds against furylfuramide, Trp-P-1 and activated Trp-P-1 were also assayed by the Ames test using S. typhimurium TA100.     

Characterization and functional analysis of a novel PCP gene BcMF5 from Chinese cabbage (Brassica campestris L. ssp. chinensis Makino)

Brassica campestris Male Fertile 5 (BcMF5), a novel member of the pollen coat protein class A (PCP-A) gene family, was identified from Brassica campestris L. ssp. chinensis Makino (Chinese cabbage-pak-choi). Temporal and spatial expression analysis showed that BcMF5 is a late-expressed PCP gene related to the process of determining pollen fertility. Functional analysis by hairpin RNA (hpRNA)-mediated RNA interference also showed that the expression of BcMF5 is inhibited, which resulted in the low germination ability of the pollen and also in an abnormality of the pollen exemplified by a collapsed germination furrow. This demonstrates that the expression of BcMF5 is closely related to the tapetum. Further, the expression profile of the BcMF5 promoter in Arabidopsis was also analyzed. This analysis indicated that the BcMF5 promoter began expression in the early stage of anther development and drove high levels of glucuronidase (GUS) expression in anthers, pollen, and the pollen tube in the late stage of pollen development, but did not drive any expression in petals, sepals, or pistils. Together with the functional analysis, the hypothesis that BcMF5 may have a sporophytic or gametophytic expression pattern is presented.