The effect of chalcones extracted from Angelica Keiskei on insulin resistance of type Ⅱ diabetes

目的 研究明日叶查尔酮对2型糖尿病骨骼肌胰岛素抵抗的影响作用及其机制。 方法 采用浸提法从明日叶中提取制备查尔酮，采用体外细胞培养和体内动物实验的方法研究明日叶查尔酮对胰岛素抵抗时骨骼肌葡萄糖代谢的影响作用及其机制。 体外细胞培养：采用高浓度胰岛素诱导的胰岛素抵抗L6细胞，分别加入低、中、高剂量依次为5μmol/L、10μmol/L、20μmol/L终浓度的明日叶查尔酮共同培养24h。采用葡萄糖氧化酶-过氧化物酶法(Glucoseoxidase-peroxidase，GOD-POD)检测培养基上清液中残余的葡萄糖含量，逆转录聚合酶链式反应方法（RT-PCR）检测PI3K、Akt、ACC...Objective To investigate the effect of chalcones extracted from Angelica keiskeion (AC) on insulin resistance of skeletal muscle with type 2 diabetes in vivo and vitro. Methods In vitro study: Differentiated L6 cell line with insulin resistance induced by high insulin was incubated with Angelica keiskei chalcones at the concentrations of 20 μmol/L, 10μmol/L and 5 μmol/L for 24 h. The contents o...学位：医学硕士院系专业：公共卫生学院_营养与食品卫生学学号：3262014115056

明日叶查尔酮对胰岛素抵抗细胞PI3K-Akt-GLUT4信号通路的影响

目的研究明日叶查尔酮(Angelica keiskei; chalcones,AC)对胰岛素抵抗L6细胞磷脂酰肌醇3激酶-蛋白激酶-葡萄糖转运体4(PI3K-Akt-GLUT4)信号通路的影响。方法采用; 高浓度胰岛素诱导的胰岛素抵抗L6细胞,分别加入5、10、20 mumol /L终浓度的明日叶查尔酮共同培养24; h,葡萄糖氧化酶法测定细胞上清液中葡萄糖含量,逆转录聚合酶链式反应方法检测细胞PI3K和Akt的mRNA表达水平,蛋白印迹法检测GLUT4和Ak; t的蛋白表达水平。结果胰岛素抵抗组细胞培养液的葡萄糖含量显著高于空白对照组(P < 0.05), 而PI3K mRNA、Akt; mRNA和GLUT4、 Akt蛋白表达水平则降低(P < 0.05);中、高剂量AC组细胞培养液葡萄糖含量均低于胰岛素抵抗组(P <; 0.05), 而PI3K mRNA、Akt mRNA和GLUT4、Akt蛋白表达水平则升高(P < 0.05); 。结论明日叶查尔酮可上调L6细胞胰岛素抵抗模型PI3K-Akt- GLUT4信号通路的表达水平,增强其对葡萄糖的摄取利用,改善胰岛素抵抗。福建省自然科学基金计划资助项

Study on the Isolation Identification and Antitumor Activity of a Phenol Derivative from Mangrove Fungus BYY-1

[作者简介] 杜希萍( 1978—) ，女，讲师，从事天然产物化学方面的研究． 通讯作者: 苏文金( 1956—) ，男，教授，从事海洋药物与食品方向研究，E-mail: wjsu@ jmu. edu. cn．[中文文摘]采用硅胶柱层析、薄层层析、Sephadex LH-20柱层析从红树内生真菌BYY-1的次级代谢产物中分离到化合物B1.通过乙酰化、NMR、HR ESI-Q-TOF MS等技术鉴定了化合物B1的结构,确定其为首次从红树内生真菌中分离到的化合物.运用MTT法和流式细胞术研究了化合物B1的抗肿瘤活性,结果表明,化合物B1可显著抑制Hela细胞的增殖[英文文摘]Compound B1 was obtained from the secondary metabolites of mangrove fungus BYY －1 using silica gel column chromatography，thin layer chromatography and Sephadex LH － 20 column chromatography．The structure of compound B1 was elucidated by acetylation，NMR and HR ESI －Q －TOF MS analysis，and compound B1 was isolated from mangrove fungi for the first time． The antitumor activity of compound B1 was studied by the methods of MTT and flow cytometric DNA analysis． The results showed compound B1 could significantly inhibit the proliferation of Hela cells with IC50 3. 3 μg /mL，and induce evident Hela cells apoptosis when compound B1 was 4 μg /mL．中央高校基本业务费资助项目(2010121092); 福建省科技计划重点项目(2011N1008);; 集美大学博士启动基金资助项目(ZQ2011004

Antitumor Activity of Mangrove Endophytic Fungi

作者简介: 郑忠辉(1956-),男, 教授[中文文摘]从红树植物秋茄、桐花树和白骨壤树皮样品中分离出125株内生真菌。采用MTT法对其体外抗肿瘤活性进行检测,发现11株红树植物内生真菌对KB(人口腔上皮癌)或HL-60(人白血病)细胞具有显著的抑制活性,占总分离菌株的8.8％。秋茄、桐花树和白骨壤的抗肿瘤活性菌株分别占供测菌株的11.6％、5.7％和4.8％。抗肿瘤活性菌株主要分布在拟青霉属及曲霉属。在所分离的菌株中,归属于拟青霉属的活性菌株占供测菌株的比例最高、活性最强。结果显示红树植物内生真菌是潜在的抗肿瘤药物资源。[英文文摘]One hundred and twenty-five strains of endophytic fungi were isolated from the inner barks of three kinds of mangrove plants,Kandelin canda,Avicennia marina and Aegiceras corniculatum.Antitumor activity was studied by the MTT assay.8.8% of endophytic fungi fermentation broths displayed cytotoxic activity on KB or HL-60 cells at and below a dilution of 1:100,11.6% cytotoxic-positive strains isolated from Kandelin canda,5.7% from Avicennia marina,and 4.8% from Aegiceras corniculatum.The active strains obtained mostly belong to Paecilomyces and Aspergillllus.Among all endophytic fungi isolated ,the genus Paecilomyces has the highest cytotoxic-positive rate and antitvmor activity.These results indicate mangrove endophytic fungi could be a promising source for antitumor bioactive agents.国家“863”计划(2001AA620401);福建省自然科学基金(C0210001

The Effects of Different Media on Antitumor Active Substances Production of Mangrove Fungus HTF3

对红树林真菌HTF3菌株在PD、GPY、YES、GPM及SYE 5种不同培养基中的生长情况、抗肿瘤活性及代谢产物的形成进行了研究,旨在了解培养基对该菌株生长及抗肿瘤活性物质合成的影响.结果显示:在不同的培养基中,HTF3菌株的生长速率不同,抗肿瘤活性及抗瘤谱也有明显差异;采用HPLC对不同培养基发酵液乙酸乙酯抽提物进行分析,发现不同抽提物的色谱峰数、色谱峰的保留时间及紫外吸收值均有较大的不同.实验结果表明:不同培养基对HTF3菌株抗肿瘤活性物质的合成有显著影响.Mangroves are special woody plant communities in the intertidal zone of tropical and subtropical coasts.Mangrove fungi growing in mangroves are attractive because a variety of chemically interesting and biologically significally substances have been purified from their secondary metabolites.Therefore,this paper studied the growth,antitumor activities and metabolites production of mangrove fungus HTF3 on potato dextrose medium,glucose peptone yeast extract medium,yeast extract sucrose medium,glucose peptone malt extract medium and starch yeast extract medium,to find out the effects of different media on the growth and antitumor active substances production of HTF3.The results showed that the growth rates of HTF3 on different solid media were different;the growth rates of HTF3 on glucose peptone yeast extract medium and starch yeast extract medium was 17.1mm/d and 11.1 mm/d;antitumor activities and antitumor specffum were markedly different in different liquid media,the IC_(50) against Raji cell of five extracts from different fermentation broth,namely A(PD),B(GPY),C(YES),D(GPM) and E(SYE) is 31.9,66.5,76,49.6 and 27.6 μg/mL respectively.The ethyl acetate extracts of the fermentation broth from different media were analysed using HPLC.The numbers of chromatographic peak,retention times and UV spectra were significant different.The study indicated diffetent media greatly affected the antitumor active substances production of HTF3.国家863计划(2001AA620401);; 福建省科技计划项目(2004N003)资

Nematicidal and Brine Shrimp Lethality of Secondary Metabolites from Marine-derived Fungi

对采集自浙江舟山海域底泥、福建省九龙江口红树林及福建省云霄县红树林共188株海洋真菌代谢产物的杀小杆线虫(Rhabditissp.)和卤虫(brine shrimp)活性进行初步研究.筛选出#164菌株代谢产物对小杆线虫显示了较好的杀害活性,其LC90小于0.5 mg/mL.#122菌株代谢产物对卤虫显示了较强的麻痹效应.#305菌株代谢产物对卤虫的LC50仅为75μg/mL.此外,对188株海洋真菌代谢产物的乙酰胆碱酯酶抑制活性进行研究,筛选出#229菌株代谢产物和#170菌株代谢产物具有较强的乙酰胆碱酯酶抑制活性,IC50分别为30μg/mL和25μg/mL,这些活性菌株具有产生杀线虫剂或乙酰胆碱酯酶抑制剂的特性.The nematicidal,brine shrimp lethality and acetylcholinesterase inhibitory activity of secondary metabolites from 188 marine-derived fungi were tested.The fungi were derived from the sediment in Zhoushan sea area and the mangrove at Yunxiao County and Jiulongjiang estuary in Fujian Province.The ethyl acetate extract of #164 culture filtrate showed strong lethal effect on nematode Rhabditis sp.at concentration of 0.5 mg/mL.Its LC_(90) was less than 0.5 mg/mL.The ethyl acetate extract of #122 culture filtrate showed strong paralyzing effect on brine shrimp at concentration of 0.5 mg/mL.It leaded to 100% mortality after treating for 2 h.And the ethyl acetate extract of #305 culture filtrate exhibited strong activity against brine shrimp at concentration of 100 μg/mL.It caused 90% mortality after treating for 24 h.Additionally,the ethyl acetate extracts of #229 and #170 showed strong inhibitory activity of acetylcholinesterase.The hyperactive acetylcholinesterase was one important character of Alzheimer's Disease symptoms.Acetylcholinesterase inhibitors were one effective approach for treating the symptoms of Alzheimer's Disease.Acetylcholinesterase inhibitors were also one important type of insecticides.It inhibited the loss of acetylcholine and impelled the overexcited state of insect's mental functions.Ultimately,it leaded the insect to death.All the bioactive marine-derived fungi had the possibility to produce nematicide or acetylcholinesterase inhibitors.国家863计划基金(2001AA620401);; 福建省科技计划(2004N003)基金资

Purification,Elucidation and Activities Study of Cytosporone B

从福建九龙江口红树林自然保护区桐花树树皮中分离出内生真菌HTF3菌株,用柱层析和正相HPLC的方法从该菌株的发酵液中纯化出一单体化合物,经1D- 、2D -NMR和ESI -MS等方法鉴定该化合物为CytosporoneB.活性检测发现它具有较强的抗肿瘤活性和较广谱的抗真菌活性,本文报道这类物质有抗肿瘤活性,作用机理值得进一步深入研究.The microorgamism is an attractive resource for chemically interesting and biologically significantly secondary metabolites.Mangroves are special woody plant communities in the intertidal zone of tropical and subtropical coasts.They are proved to be natural microorganisms and new metabolites storage.An endophytic fungus,Dothiorella sp.,strain HTF3 was isolated from mangrove plant Avicennia marina at the estuary of Jiulong River,Fujian Province.Its fermentation broth was extracted with ethyl acetate and the extract was subjected to column chromatography on silica gel and HPLC.A compound was purified,its structure was elucidated by ESI,1D- and 2D-NMR spectra as Cytosporone B.Its IC_(50) values were 4 μg/mL and 0.062 5 μg/mL against human epidermal carcinoma of oral cavity,KB cell line and human Burkitt’s lymphoma,Raji cell line,respectively.Furthermore,it showed broad activities against fungi,the MIC against Aspergillus niger、Trichoderma sp.and Fusarium sp were 0.125 mg/mL、62.5 μg/mL and 62.5 μg/mL,respectively.Its high antitumor activities and broad anti-fungi spectra indicates that it is worthy further research.国家 863计划 ( 2001AA620401 );; 福建省科技计划项目(2004N003)资

Antimicrobial and Antitumor Activities of Marine Fungi Isolated from Sediment in Deep Sea

从西太平洋近赤道区采集到的18个深海沉积物样品中分离到83株海洋真菌,采用滤纸片琼脂扩散法和MTT法分别对其发酵液乙酸乙酯抽提物的抗菌、抗肿瘤活性进行筛选.结果显示,有37株菌株对至少一种指示菌有抑制作用;有29株菌株对KB或Raji肿瘤细胞具有显著的抑制活性,分别占总供测菌株的44.6%和34.9%.在抗肿瘤活性菌株中,WP-M-1、DY-G-2、DY-C-2、WP-K-3和WP-Q-2等5株菌具有很高的活性,其发酵液乙酸乙酯抽提物对Raji细胞的IC50分别为5.000、1.064、2.796、1.920、0.520μg/mL.研究结果表明,海洋沉积物来源的真菌中蕴藏着丰富的抗菌及抗肿瘤活性代谢产物产生菌,是开发抗菌和抗肿瘤药物的宝贵资源.Marine organisms are a rich source of structurally novel and biologically active metabolites.So far,many chemically unique compounds of marine origin with different biologically activity have been isolated and a number of them are under investigation and/or are being developed as new pharmaceuticals.In this report,we describe the screening of ethyl acetate extracts of eighty-three strains of marine sediment fungi,collected from west of the Pacific Ocean beside the equator,for antimicrobial and antitumor activities.As a result,thirtyseven strains showed inhibition against one or more of those tested microorganism,namely EC,SA,BS,CA,ANS,while twenty-nine strains of fungi demonstrated antitumor activity againnst KB cell or Raji cell respectively.The ratio of such activity fungi to total tested fungi was 44.6%,34.9%.Furthermore,five fungal extracts,namely WP-M-1、DY-G-2、DY-C-2、WP-K-3 and P-Q-2,were chosen to study the IC_(50) against Raji cell,the value of them was 5.000,1.064,2.796,1.920,0.520μg/mL respectively.These results indicated that marine sediment fungi would be a promossing resourse of antimicrobial and antitumor medicament.国家863计划(2003AA624010);; 福建省科技计划项目(2004N003)资

Isolation and structure elucidation of lipophilic cytotoxic compounds produced by a marine actinomycetes, N350

海洋微生物是当前抗肿瘤药物开发的重要新资源.应用硅胶柱层析结合活性跟踪分离的方法,从海洋放线菌菌株N350的脂溶性发酵产物中纯化并鉴定了4个化合物,分别为反式-13-二十二碳单烯酰胺,7,4'-二羟基异黄酮、6-甲氧基-7,4'-二羟基异黄酮及β-胡萝卜甙.其中,反式-13-二十二碳单烯酰胺、7,4'-二羟基异黄酮及6-甲氧基-7,4'-二羟基异黄酮具有抗肿瘤活性,抑制人口腔上皮癌KB细胞生长的ID50分别为15,8 1,5 2μg/cm3,抑制人白血病细胞HL-60生长的ID50分别为7,2 8,1 9μg/cm3.这些化合物均首次在海洋放线菌中得到分离.Marine microorganism, which can be easily reproduced in large scale in industry, is being exploited for new antitumor drugs with unique structure and excellent bioactivity. Four lipophilic cytotoxic compounds produced by a marine actinomycetes, strain N350, were purified by bioassay-guided isolation, and identified as cis-13-docosenmamide, 7, 4′-dihydroxyisoflavone, 6-methoxy-7, 4′-dihydroxyisoflavone, and β-sitosterol-β-D-glucopyranoside, respectively. Cis-13-docosenamide, 7, 4′-dihydroxyisoflavone and 6-methoxy-7, 4′-dihydroxyisoflavone demonstrated significant cytotoxic activity in vitro. Against human leukemia cell line, HL-60, their IC_(50) was 2.8, 1.9, 7 μg/cm~3, respectively, and against human epidermal carcinoma of oral cavity, KB cell line, IC_(50) was 8.1, 5.2, 15 μg/cm~3, respectively. All these compounds were, for the first time, isolated from the products of marine actinomycetes.国家"863"计划资助项目(2001AA620401);; 国家自然科学基金资助项目(0030007024);; 福建省自然科学基金资助项目(F99032)