Anticancer and antiviral activities of Youngia japonica (L.) DC (Asteraceae, Compositae)

Aqueous and ethanol extracts of Youngia japonica (also known as Oriental hawksbeard) were tested in vitro for anti-tumor activity against three cell lines, human promyelocytic leukaemia (HL-60), human myelogenous leukaemia (chronic K-562) and mouse Sarcoma 180 (S-180), and for antiviral activity against respiratory syncytial virus (RSV), influenza A virus (Flu A) and herpes simplex virus type 1 (HSV-1) by cytopathic effect (CPE) reduction assay. Hot water extract of Youngia japonica inhibited cell proliferation and growth of all cancer cell lines to various extent. K-562 cells were the most sensitive to the extract whereas S-180 cells were the least. It did not show any significant cytotoxic effects on normal mammalian Vero cells up to the concentration of 450 microg/mL. The ethanol extract of whole plant of Youngia japonica exhibited antiviral activity against RSV cultured in HEp-2 cells, but did not have any activity against Flu A and HSV-1. Two partially purified fractions (Fr.10 and Fr.11) from the 95% ethanol extract exhibited significant anti-RSV with 50% inhibitory concentration (IC50) in the range of 3.0-6.0 microg/mL. The ratio of the viral titer reduction in the absence (viral control) and presence of the maximal non-cytotoxic concentration (MNCC) of the Fr.10 and Fr.11 was both estimated to be 1 x 10(4) (RF, viral titer reduction factors), indicating that their anti-RSV activity was high enough to justify for further analysis. Our preliminary analysis showed that the antiviral ingredients were likely to contain phenolic compounds including tannins by chemical tests.     

基于在体循环肠灌流模型分析羊耳菊提取物的肠吸收特性

目的:考察羊耳菊提取物在大鼠肠内的吸收特征。方法:采用大鼠在体循环肠灌流模型,选择羊耳菊提取物中东莨菪苷等9种代表性成分为考察对象,建立其UPLC-MS/MS并测量各成分的累计吸收量,探究药物质量浓度,胆汁,P-糖蛋白(P-gp)抑制剂及不同肠段对羊耳菊提取物中东莨菪苷等成分肠吸收的影响,阐明各成分在不同肠段的吸收特征。结果:东莨菪苷在高质量浓度条件下存在饱和现象,提示可能的吸收方式为主动转运,而1,3-O-二咖啡酰基奎宁酸等其余8种成分则无明显高浓度饱和现象,提示这8种成分可能的吸收方式均为被动扩散。木犀草苷,1,3-O-二咖啡酰基奎宁酸,3,4-O-二咖啡酰基奎宁酸和4,5-O-二咖啡酰基奎宁酸的主要吸收部位在空肠,东莨菪苷和3,5-O-二咖啡酰基奎宁酸的主要吸收部位在回肠,绿原酸、新绿原酸和隐绿原酸的主要吸收部位在十二指肠。羊耳菊提取物中东莨菪苷等9种成分的肠吸收均受到p H和胆汁的影响。结论:羊耳菊提取物中9种成分在小肠均有吸收,但各成分的吸收速率、最佳吸收部位及吸收机制不尽相同。     

Cytotoxicity and in vitro susceptibility of Entamoeba histolytica to Morinda morindoides leaf extracts and its isolated constituents

An aqueous decotion (dried extract), an 80% MeOH extract from Morinda morindoides leaves, and 10 flavonoids and 4 iridoids isolated from the 80% MeOH extract were evaluated in vitro for their potential antiamoebic activity and their cytotoxic effect against MT-4 cells. Results indicated that the aqueous decoction and the 80% MeOH extract exhibited an interesting antiamoebic activity with IC(50) values of 3.1 +/- 1.7 and 1.7 +/- 0.6 microg/ml, respectively. Apigenin-7-O-glucoside and luteolin-7-O-glucoside exhibited a moderate antiamoebic activity with IC(50) values of 22.3 +/- 3.2 and 37.4 +/- 2.7 microg/ml, respectively. Kaempferol (IC(50) = 10.3 +/- 2.3 microg/ml), apigenin (IC(50) = 12.7 +/- 4.3 microg/ml), and luteolin (IC(50) = 17.8 +/- 4.3 microg/ml) showed a more pronounced activity than their corresponding glycosides. All tested iridoids displayed a very good activity with IC(50) values less than 10 microg/ml. The most active iridoids were epoxygaertneroside (IC(50) = 1.3 +/- 0.4 microg/ml) and methoxygaertneroside (IC(50) = 2.3. +/- 0.7), followed by gaertneroside and gaertneric acid with IC(50) values of 4.3 +/- 1.8 and 7.1 +/- 1.4 microg/ml, respectively. Except quercetin and quercetin-7,4'-dimethylether which have shown a cytotoxic effect with IC(50) ranging from 14 to 22 microg/ml. No correlation could be deduced between the observed antiamoebic and cytotoxic activity of these tested samples. A structure-activity relationship for isolated compounds is discussed. These findings support the medicinal report for the traditional use of Morinda morindoides leaves for the treatment of amoebiasis.     

旱柳叶中三种黄酮类成分及水杨苷含量的高效液相色谱法测定

目的建立同时测定旱柳叶中木樨草素-7-O-葡萄糖苷、水杨苷、芹菜素-3’-氧乙基-7-O-葡萄糖苷及杨梅酮含量的高效液相色谱(HPLC)方法。方法 Nucleosil C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-水(5∶95)-(30∶70)为梯度流动相;流速为1.0 ml.min-1;检测波长为246 nm;柱温为25℃。结果木樨草素-7-O-葡萄糖苷在0.02～0.1 mg范围内与峰面积呈良好线性关系,R=0.999 0,平均回收率为97.81%,RSD为2.93%;水杨苷在0.03～0.15 mg范围内与峰面积呈良好线性关系,R=0.998 8,平均回收率为97.54%,RSD为1.62%;芹菜素-3’-氧乙基-7-O-葡萄糖苷在0.008～0.04 mg范围内与峰面积呈良好线性关系,R=0.999 1,平均回收率为97.41%,RSD为1.37%;杨梅酮在0.04～0.2 mg范围内与峰面积呈良好线性关系,R=0.9977,平均回收率为97.91%,RSD为1.03%。结论此法简便快速,结果准确可靠、重复性好,可用于药材中木樨草素-7-O-葡萄糖苷、水杨苷、芹菜素-3’-氧乙基-7-O-葡萄糖苷及杨梅酮含量的同时测定。     

Angiotensin converting enzyme inhibitors from Cuscuta japonica Choisy

Bioassay-guided fractionation of the EtOAc-soluble extract of Cuscuta japonica afforded 3,5-Di-O-caffeoylquinic acid (1). methyl 3,5-Di-O-caffeoylquinate (2). 3,4-Di-O-caffeoylquinic acid (3). and methyl 3,4-Di-O-caffeoylquinate (4). Purification of these compounds was conducted with the application of various chromatographic methods. The structures of the compounds were elucidated on the basis of MS and NMR data analysis. Compounds 1-4 inhibited the angiotensin I converting enzyme activity in a dose-dependent manner. Compounds 1-4 showed the 50% inhibitory concentration values of 596, 483, 534, and 460 micro M, respectively. The presence of these active components may be responsible, at least in part, for the antihypertensive action of traditional crude drug Cuscuta Semen.     

浮萍的化学成分研究

从中药浮萍的正品紫萍的乙醇提取物中分离出４个黄酮化合物,经理化常数和光谱鉴定其结构,分别为芹菜素（ａｐｉｇｅｎｉｎ）,木犀草素（ｌｕｔｅｏｌｉｎ）芹菜素－７－Ｏ－葡萄糖革（ａｐｅｇｅｎｉｎ－７－Ｏ－ｇｌｕｃｏｓｉｄｅ）和水犀草素－７－Ｏ－葡萄苷（ｌｅｕｔｅｏｌｉｎ－７－Ｏ－ｇｌｕｃｏｓｉｄｅ）它们是紫萍中的主要化学成分。     

口炎清中酚酸类成分研究

目的:研究口炎清流膏中的酚酸类成分.方法:采用薄层、硅胶柱和大孔树脂柱等色谱和制备HPLC方法进行化合物的分离,通过HPLC分析及MS、1 H-NMR、13 C-NMR等技术进行化合物的结构鉴定.结果:从口炎清流膏中分离得到4个酚酸类成分:咖啡酸(1)、绿原酸(2)、3,5-二咖啡酰奎宁酸(3)和3,4-二咖啡酰奎宁酸(4).结论:首次对中药口炎清复方化学成分进行研究,4个酚酸类成分均来自于君药山银花.     

野菊花心血管活性部位化学成分的研究

目的:研究菊科植物野菊花心血管活性部位中的主要化学成分.方法:采用多种色谱方法进行分离纯化,通过波谱方法结合文献对照鉴定化合物结构.结果:从野菊花心血管活性部位中分离并鉴定了12个化合物,包括7个黄酮类化合物(1～7),5个苯丙素类化合物(8～12),分别鉴定为(2S)-圣草酚-7-O-β-D-吡喃葡萄糖醛酸苷(1),(2S)-圣草酚-7-O-β-D-吡喃葡萄糖苷(2),(2S)-橙皮素-7-O-β-D-吡喃葡萄糖醛酸苷(3),木犀草素-7-O-β-D-吡喃葡萄糖苷(4),木犀草素-7-O-β-D-吡喃葡萄糖醛酸苷(5),香叶木素-7-O-β-D-吡喃葡萄糖醛酸苷(6),槲皮素-7-O-β-D-呲喃葡萄糖苷(7),3,5-二咖啡酰奎宁酸甲酯(8),3,5-二咖啡酰奎宁酸(9),3,5-二顺式咖啡酰奎宁酸(10),1,5-二咖啡酰奎宁酸(11),1,3-二咖啡酰奎宁酸(12).结论:野菊花心血管活性部位中主要成分为黄酮类化合物,共5个化合物分别为2～3,7～8,10;共10个化合物分别为2～3,5～12系首次从该种植物分离得到.     

Anti-inflammatory activity of extract and fractions from Nepeta sibthorpii Bentham

Several species of Nepeta genus are utilized in folk medicine for treatment of contusions, rheumatic pains, fever, cutaneous eruptions. Some species are employed for their anti-inflammatory properties. In this paper, we report the results of phytochemical studies on aerial parts of Nepeta sibthorpii Bentham (Lamiaceae), an endemic plant of Greece. The bioassay-guided fractionation of methanol extract led to the isolation of ursolic acid and polyphenol fraction. By HPLC, we determined some phenolics: chlorogenic acid (0.315 mg/g) and the flavonoids rutin (0.091 mg/g), luteolin-7-O-glucoside (0.387 mg/g) and a luteolin derivative. We assayed the radical scavenging activity of Nepeta sibthorpii methanol extract by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. Moreover, we studied the anti-inflammatory activity of Nepeta sibthorpii methanol extract (50 mg/kg, os), ursolic acid and polyphenol fraction (dose corresponding to 50 mg/kg of methanol extract, os) in the carrageenan-induced paw oedema in rat. In this experimental model, we observed a significant inhibition of paw oedema. We suppose that the anti-inflammatory effect of methanol extract could be related to the free radical scavenging activity and that it depends on a synergic action of all the components of the methanol extract, even if ursolic acid can be considered the main responsible for this activity.     

Antibacterial activities and cytotoxicity of terpenoids isolated from Spirostachys africana

Spirostachys africana Sond. stem bark is used traditionally for the treatment of diarrhoea and dysentery in Limpopo Province of South Africa. Bioassay-guided fractionation of ethanolic extract from bark of Spirostachys africana led to the isolation of four known compounds, two triterpenoids, compound 1 [d-Friedoolean-14-en-oic acid (3-acetyl aleuritolic acid)] and compound 2 (Lupeol), and two diterpenes, compound 3 [ent-2,6alpha-dihydroxy-norbeyer-1,4,15-trien-3-one (diosphenol 2)] and compound 4 (ent-3beta-hydroxy-beyer-15-ene-2-one). Isolated compounds were tested for antibacterial activity using micro-dilution method. Compound 1, exhibited minimum inhibitory concentration (MIC) of 50 microg/ml against Staphylococcus aureus, Salmonella typhy, Vibrio cholera, Escherichia coli and Shigella dysentery. Compound 2 was not active against all tested microorganisms at 200 microg/ml, which was the highest concentration tested. At this concentration, all four compounds were not active against Shigella sonnei. Cytotoxicity of ethanol crude extracts and isolated compounds from Spirostachys africana was determined using the sodium-2,3-bis-[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT) assay on Vero cells. Compounds 2 and 3, isolated from Spirostachys africana, had up to three times higher [50% inhibitory concentration (IC(50) values; 300.9 and 308.9 microg/ml)] than the ethanol crude extracts (102.8 microg/ml) suggesting higher toxicity of the crude extract as compared to these two compounds. In contrast, compounds 1 and 4 were not cytotoxic to Vero cell lines (African green monkey) in vitro at the concentrations tested (IC(50)>400 microg/ml). This is the first report on the antibacterial activity and cytotoxicity of purified compounds from Spirostachys africana.     

Free radical scavenging and membrane protective effects of methanol extracts from Anthriscus cerefolium L. (Hoffm.) and Petroselinum crispum(Mill.) nym. ex A.W. Hill

The antilipoperoxidant activity of Anthriscus cerefolium L. (Hoffm.), chervil, Petroselinum crispum (Mill.) Nym. ex A. W. Hill., parsley extracts were evaluated with ascorbic acid induced lipid peroxidation on rat brain homogenates. These results are completed by the antiradical potential of these extracts against a solution of OH. radical. In all cases luteolin-7-O-glucoside was used as a reference material.     

Metabolite profiling and chemopreventive bioactivity of plant extracts from Bidens pilosa

Bidens pilosa has been used as a folk medicine in various medications and as a popular ingredient in herb teas. Chemopreventive activities of crude and fractionated plant extracts of Bidens pilosa were evaluated in this study. Ethyl acetate and butanolic fractions, partitioned from the total crude extract of Bidens pilosa, exhibited significant scavenging free radical activity (IC(50) values approximately = with 14-17 microg/mL) comparable to that of alpha-tocopherol. Strong effects on the inhibition of LPS-mediated nitric oxide production in RAW 264.7 cells were also observed for the EA and BuOH fractions. Detectable cytotoxicity on RAW 264.7 cells, however, was observed for the EA fraction at a dose >100 microg/ml. The metabolite profile and major constituents of the BuOH fraction were studied and characterized using various spectroscopic analyses. A new compound, heptanyl 2-O-beta-xylofuranosyl-(1-->6)-beta-glucopyranoside (1), and eight phenolic compounds, namely quercetin 3-O-rabinobioside (2), quercetin 3-O-rutinoside (3), chlorogenic acid (4), 3,4-di-O-caffeoylquinic acid (5), 3,5-di-O-caffeoylquinic acid (6), 4,5-di-O-caffeoylquinic acid (7), jacein (8), centaurein (9) were for the first time isolated from Bidens pilosa. Compounds 2-7 are the major antioxidative constituents in the Bidens pilosa extract.     

Inhibition of angiotensin converting enzyme (ACE) by flavonoids isolated from Ailanthus excelsa (Roxb) (Simaroubaceae)

In our screening program for antihypertensive properties of plants, the leaves of Ailanthus excelsa (Roxb), a plant used in Egyptian traditional medicine, were analysed. Chromatographic separation of A. excelsa MeOH extract yielded six flavonoids for the first time from this species, namely apigenin, luteolin, kaempferol-3-O-alpha-arabinopyranoside, kaempferol-3-O-beta-galactopyranoside, quercetin-3-O-alpha-arabinopyranoside and luteolin-7-O-beta-glucopyranoside. The in vitro hypotensive activities of the MeOH extract and the isolated compounds were elucidated. All the flavonoids tested exhibited ACE inhibitory activity, in particular the most active compound was kaempferol-3-O-beta-galactopyranoside with an IC(50) value of 260 microm.     

两色金鸡菊提取物大剂量给药致小鼠死亡化学成分的研究

目的比较两色金鸡菊各种提取物对小鼠的急性毒性作用,分析对大剂量给药致小鼠死亡贡献最大的成分,为后期研究提供安全性数据并优化提取工艺。方法分别测定各提取物对小鼠的最大给药量或最大耐受量(MTD),用Bliss法测定半数致死量(LD50),记录小鼠死亡情况和体质量变化情况;高效液相色谱(HPLC)法测定各提取物指纹图谱,根据各物质吸收峰,结合多元线性回归,分析对大剂量给药致小鼠死亡贡献最大的成分。结果两色金鸡菊喷雾干燥水提物(SD)、醋酸乙酯萃余组分(AR)最大给药量均为36 g/kg,真空干燥水提物(VD)MTD为26 g/kg,乙醇提取物(ETE)LD50(95%可信限)为19.565(17.558~21.734)g/kg,醋酸乙酯萃取组分(AC)LD50(95%可信限)为16.414(13.987~34.725)g/kg;3,5-二咖啡酰奎宁酸可能是对大剂量给药致小鼠死亡贡献最大的成分。结论 ETE和AC大剂量给药会致小鼠死亡,3,5-二咖啡酰奎宁酸可能是对两色金鸡菊提取物大剂量给药致小鼠死亡贡献最大的成分。     

野菊不同部位绿原酸和3,5-二咖啡酰奎尼酸的含量测定

目的:检测湖南衡阳产野菊不同部位(叶、花、花蕾、嫩茎及老茎)中绿原酸和3,5-二咖啡酰奎尼酸的含量.方法:以80％的乙醇为溶剂,用超声法提取野菊不同部位中的绿原酸和3,5-二咖啡酰奎尼酸.采用高效液相色谱法检测绿原酸和3,5-二咖啡酰奎尼酸的含量,以0.05％磷酸水溶液-乙腈为流动相,梯度洗脱,流速1.0 mL·min-1,检测波长327 nm.结果:绿原酸和3,5-二咖啡酰奎尼酸的质量浓度在10～100 mg·L-1(r =0.996,r =0.992)与蜂面积呈良好的线性关系(n=5),平均回收率(n=6)分别为98.5％,99.7％.野菊叶、花、花蕾、嫩茎及老茎中绿原酸的含量分别为1.08％,0.44％,0.39％,0.58％及0.52％;3,5-二咖啡酰奎尼酸的含量分别为1.66％,1.01％,0.82％,0.93％,0.64％,其中野菊叶中绿原酸和3,5-二咖啡酰奎尼酸的含量最高.结论:野菊不同部位均含有绿原酸和3,5-二咖啡酰奎尼酸,但它们的含量存在明显差异.     

Immunomodulatory effect of Glossogyne tenuifolia in murine peritoneal macrophages and splenocytes

Glossogyne tenuifolia Cass., a medicinal plant native to Taiwan, is traditionally used as an anti-inflammatory remedy. Oleanolic acid and luteolin-7-glucoside have been previously identified as active components of Glossogyne tenuifolia in the murine macrophage-like cell line, RAW264.7. Current study investigates the effect and mechanism of the ethanol extract of Glossogyne tenuifolia (GT) and its major constituents on the release of inflammatory mediators in activated elicited murine peritoneal macrophages and splenocytes. Our results showed that GT (up to 0.15 mg/ml) inhibited the production of proinflammatory mediators, TNF-alpha, IL-1beta, IL-6, nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in LPS-activated macrophages, and IFN-gamma in PHA-activated splenocytes. GT also inhibited LPS-activated murine iNOS and COX-2 promoter activities in transiently transfected RAW264.7 cells. The major constituents, oleanolic acid and luteolin-7-glucoside, as well as its aglycone, luteolin, inhibited the release of NO, PGE(2), TNF-alpha and IL-1beta in activated peritoneal macrophages. However, only luteolin-7-glucoside and luteolin were able to reduce IFN-gamma release in PHA-stimulated splenocytes. To further investigate the possible mechanisms that interfere with LPS- and PHA-signaling, this study focused on nuclear factor-kappaB activation signaling pathways. Our results demonstrate that GT (0.075-0.15 mg/ml) treatment reduces nuclear factor-kappaB (NF-kappaB) DNA binding activity, as demonstrated by electrophoretic mobility shift assay (EMSA). Collectively, the results suggest that GT inhibits proinflammatory mediator synthesis in activated murine peritoneal macrophages and splenocytes, in part through NF-kappaB-dependent pathways.     

Phenolic substances from Phagnalon rupestre protect against 2,4,6-trinitrochlorobenzene-induced contact hypersensitivity

2-isoprenylhydroquinone-1-glucoside (1), 3,5-dicaffeoylquinic acid (2), and 3,5-dicaffeoylquinic acid methyl ester (3), isolated from Phagnalon rupestre, improved the contact hypersensitivity response to 2,4,6-trinitrochlorobenzene in mice. These phenolics reduced ear swelling and IL-1β content by 50% 24 h after challenge; in addition, 2 inhibited tumor necrosis factor-α by 53%. All three compounds also reduced interleukin-2 content by 50% 72 h after challenge. Both 2 and 3 inhibited metalloproteinase-9 levels in the skin lesions by 66% and 41%, respectively, and lowered cyclooxygenase-2 expression by 44% and 49%, respectively, at 24 h. Moreover, 2 was effective against atopic dermatitis induced by repeated application of 2,4,6-trinitrochlorobenzene; it attenuated edema by over 40% from day 7 and inhibited inflammatory cell infiltration by 44% at day 22. In addition, 1-3 reduced metalloproteinase-9 expression in a dose-dependent manner in macrophages RAW 264.7 stimulated with lipopolysaccharide. Thus, compounds 2 and 3 were found to exhibit a greater activity against contact hypersensitivity than 1.     

口炎清中酚酸类成分研究

目的：研究口炎清流膏中的酚酸类成分。方法：采用薄层、硅胶柱和大孔树脂柱等色谱和制备HPLC方法进行化合物的分离,通过HPLC分析及MS、^1H-NMR、^13C-NMR等技术进行化合物的结构鉴定。结果：从口炎清流膏中分离得到4个酚酸类成分：咖啡酸（1）、绿原酸（2）、3,5-二咖啡酰奎宁酸（3）和3,4-二咖啡酰奎宁酸（4）。结论：首次对中药口炎清复方化学成分进行研究,4个酚酸类成分均来自于君药山银花。     

Cytotoxic constituents of roots of Chaerophyllum hirsutum

Two new lignan derivatives (1 and 2), along with the known scopoletin (3), methyl caffeate (4), falcarindiol (5), 3,5-dicaffeoylquinic acid, 3,5-dicaffeoylquinic methyl ester, chlorogenic acid, p-hydroxyphenylethyl-trans-ferulate, and vanillin, were isolated from extracts of the roots of Chaerophyllum hirsutum. Structure elucidation of the new compounds was carried out by 1D and 2D NMR experiments and by HRMS analysis. Several of the isolated compounds were tested for their cytotoxic activity against the HL-60, HT-1080, LoVo, and LoVo/Doxo cell lines.     

苦碟子药材及其注射液中化学成分的HPLC-ESI-MSn分析

应用 HPLC-ESI-MSⁿ建立1种苦碟子药材及其注射液的快速定性分析方法。通过与对照品保留时间、多级质谱信息对照,快速定性分析苦碟子药材及其注射液中的化学成分。共鉴定出33个化合物,核苷类3个、有机酸类16个、黄酮类7个以及倍半萜内酯类7个,其中通过对照品比对准确鉴定了12个化合物,11个化合物为首次在苦碟子中发现。结合已知化合物的裂解规律和未知化合物的多级质谱信息,对5个同分异构体的化合物结构进行了预测。所建立的苦碟子 HPLC-ESI-MSⁿ定性分析方法简单、快速、高效,可以为苦碟子的质量控制和评价提供有效依据。