芸香中香豆素类化学成分研究

目的研究中药芸香(Ruta graveolens L.)中香豆素类化学成分,并探讨其抗炎、抗凝活性。方法本实验利用各种色谱分离技术,结合核磁共振波谱及质谱联用技术对分离得到的单体化合物进行结构鉴定。CCK-8法筛选出无明显细胞毒性的化合物,用于对脂多糖诱导小鼠巨噬细胞(RAW 264.7)释放NO的检测,进行抗炎活性筛选,通过凝血酶原时间的测试进行抗凝活性筛选。结果从芸香提取物中共鉴定出10个香豆素类化合物,分别为补骨脂素(1)、花椒毒素(2)、chalepensin(3)、rutamontine(4)、3-(1',1',dimethyl allyl)-herniarin(5)、佛手苷内酯(6)、异莨菪亭(7)、异嗪皮啶(8)、去甲基软木花椒素(9)、Chalepin(10)。对10个化合物的抗炎、抗凝进行活性评价,结果显示,化合物2、4、9在20μmol/L时对脂多糖(lipopolysaccharide,LPS)诱导的NO生成有较好的抑制作用,抑制率分别为54.3%±1.7%、51.2%±5.5%、52.3%±1.7%;10个化合物在浓度为1.0 mmol/L条件下,均未表现出抗凝活性。结论从芸香提取物中分离得到10个香豆素类单体化合物,化合物3~5、7~10为首次从芸香中分离得到。该研究结果丰富了芸香的化学成分和生物活性研究,为芸香抗炎活性的开发和利用提供了理论基础。

中药芸香中(+)/(-)-Rutabenzofuran A和(+)/(-)-Rutabenzofuran B的抗肝炎活性评价

目的研究中药芸香中分离鉴定的两对Z/E异构化的苯并呋喃香豆素对映异构体(+)/(-)-Rutabenzofuran A和(+)/(-)-Rutabenzofuran B对肝脏的保护作用。方法以(+)/(-)-Rutabenzofuran A和(+)/(-)-Rutabenzofuran B为研究对象,采用脂多糖(LPS)100 ng·mL^(-1)建立HepG2细胞的炎症模型,对4个化合物的抗肝炎活性进行初步评估。结果在浓度10、20和40μmol·L^(-1)下,4个化合物可以剂量依赖性地降低谷丙转氨酶(ALT)和谷草转氨酶(AST)水平及肿瘤坏死因子(TNF)-α水平,升高白细胞介素(IL-4)和IL^(-1)0水平。结论芸香通过抑制TNF-α和促进IL-4及IL^(-1)0的产生发挥抗肝炎活性,且具有剂量依赖性,本研究为中药芸香的临床应用及药用开发提供充分的理论依据。

Investigation on Species of Diseases and Pests in Pelargonium graveolens L.Her and Control Technique

[ Objective] The study aimed to confirm species and damage status of the diseases and pests in Pelargonium graveolens L. Her, and explore the control technique of the main pests. [Method] Investigation of pests and diseases was conducted in main growing base of P. graveolens in Guangxi Province during 2007 -2008, and randomized block design was adopted to conduct field efficacy trials. [-Result] There were 22 kinds of pests and 5 kinds of diseases causing damage on P. graveolens, the occurrence peak of main pest P. litura were in June and September. Field control test showed that 5% hexaflumuron SC and 1% emamectin microcapsule suspension had relatively high control effect on P. litura, the control effect all over 88.58% after spraying for 7 d, and the persistent effect was about 14 d. [ Conclusion] The study would provide useful support for disease and oest control in standardized cultivation of P. oraveolens.

Vasorelaxant activity of extracts obtained from Apium graveolens:Possible source for vasorelaxant molecules isolation with potential antihypertensive effect

Objective:To investigate the vasorelaxant effect of organic extracts from Apium graveolens(A.graveolens)which is a part of a group of plants subjected to pharmacological and phytochemical study with the purpose of offering it as an ideal source for obtaining lead compounds for designing new therapeutic agents with potential vasorelaxant and antihypertensive effects.Methods:An ex vivo method was employed to assess the vasorelaxant activity.This consisted of using rat aortic rings with and without endothelium precontracted with norepinephrine.Results:All extracts caused concentration-dependent relaxation in precontracted aortic rings with and without endothelium;the most active extracts were Dichloromethane and Ethyl Acetate extracts from A.graveolens.These results suggested that secondary metabolites responsible for the vasorelaxant activity belong to a group of compounds of medium polarity.Also,our evidence showed that effect induced by dichloromethane and ethyl acetate extracts from A.graveolens is mediated probably by calcium antagonism.Conclusions:A.graveolens represents an ideal source for obtaining lead compounds for designing new therapeutic agents with potential vasorelaxant and antihypertensive effects.

Anxiolytic and free radical scavenging potential of Chinese celery(Apium graveolens) extract in mice

Objective: To elucidate the anxiolytic and free radical scavenging effect of methanolic extract of Apium graveolens(A. graveolens) in adult C57BL/6 mice.Methods: Sixty male mice were divided into 6 groups: control, vehicle, positive control and A. graveolens(125, 250 and 500 mg/kg). Different behavioral models of elevated plus maze, open field, light/dark, hole-board and pentobarbital-induced sleep were used to assess anxiety-like behavior. Biochemical parameters including monoamine oxidase-A(MAO-A) activity, lipid peroxidation, % inhibition of superoxide anion and glutathione peroxidase activity were measured. Histologic studies were also examined.Results: Mice receiving various doses of A. graveolens(125, 250 and 500 mg/kg)showed an alleviation of anxiety-like behavior as evidenced by the battery of behavioral tests. Likewise, A. graveolens treatment was found to significantly decrease MAO-A activity, lipid peroxidation as well as cause a significant increase of % inhibition of superoxide anion and glutathione peroxidase activity in both cortex and striatum. The total number of survival neurons found in the frontal cortex and striatum was significantly higher than that of the vehicle-treated group.Conclusions: Taken together, we showed that A. graveolens improve the behavioral changes which might be related to the inhibition of free radicals and modulation of MAOA activity resulting in an increased number of survival neurons. Our findings suggest the therapeutic potential of A. graveolens in the treatment of anxiety.

Bioadsorption of Pb(II) onto <i>Anethum</i><i>graveolens</i>from Contaminated Wastewater: Equilibrium and Kinetic Studies

In the present study we reported the feasibility of the Anethum graveolens as biosorbent to remove Pb(II) from aqueous solutions. Anethum graveolens was characterized by scanning electron microscopy and elemental analysis. The ability of Anethum graveolens to adsorb Pb(II) was investigated by using batch adsorption procedure. The effects such as pH, contact time, adsorbate concentration and biosorbent dosage on the adsorption capacity were studied. The experimental data were analysed using various adsorption kinetic models viz., the pseudo-first and second-order equations, Bangham’s equation, intraparticle diffusion and Elovich models. Results show that the pseudo-second-order equation provides the best correlation for the biosorption process. The equilibrium nature of Pb(II) adsorption at 30℃ has been described by the Langmuir, Freundlich, Temkin and Redlich-Peterson isotherm models. The equilibrium data fit well on Langmuir isotherm. The monolayer adsorption capacity of Pb(II) onto Anethum graveolens as obtained from Langmuir isotherm at 30℃ was found to be 303 mg/g. This high adsorption capacity of Anethum graveolens places this biosorbent as one of the best adsorbents for removal of Pb(II) from aqueous effluents.

Somatic embryogenesis,synthetic seed preparation and plant regeneration in Apium graveolens var.Dulce pers

On the MS medium supplemented with 2 ppm 2,4-D,calli were induced after 4-6weeks from the petioles of an American celery plant （Apium graveolens var.Dulce pers.cv.Florida）.Suspension culture was started from the calli in a hormone-free liquid MS medium on agyratory shaker at 110 rpm,and kept at 26℃.To stimulate cell division and dedifferentiation,thesubcultures were conducted for 7 days each on the same medium.The liquid suspension containingsingle cells,cell aggregates,and somatic embryos in different stages were screened 2-3 weekslater and 1.0-1.5mm somatic embryos were obtained.These embryos were encapsulated withsodium alginate by dropping-bead method and solidified with 0.1mol CaCl2,.These synthetic seedsgerminated and developed well into seedlings in the sterilized vermiculite substrate.

Apium graveolens L. accelerating differentiation of neural stem cells in vitro

To evaluate the action of Apium graveolens L. on the growth and differentiation of neural stem cells （NSCs）, cells culture and animal experiment were performed. NSCs were isolated from the striatum of SD rat embryo were cultured in the medium containing aqueous extract from Apium graveolens L（AAG）.and/or the serum derived from mice treated with Apium graveolens L （SAG）. The results show that AAG promoted the survival and growth of NSCs in low concentration. Apium graveolens L. leaves aqueous extract promoted the proliferation of NSCs in relatively high concentration. SAG significantly accelerated the differentiation of NSCs.

Some pharmacological studies on the methanolic extract of <i>Inula graveolense</i>L.

Inula graveolens L. is widely used in Iraq for the treatment of rheumatic fever, infant convulsions, toothache, blood sugar, and also to dissolve internal blood clots, and to aid digestion. However, the efficacy and mechanisms of action of the plant remain unclear. Therefore, the objective of the present study was to investigate the pharmacological effects of the methanolic extract (MEIG) of this plant belonging to compositae family. Anti-diarrheal and antipyretic activities of the extract were examined in rats. Anti-inflammatory and antinociceptive were studied in mice. At the doses of 200 (P in vitro protein anti-denaturation using Bovine serum albumin and anti-platelet aggregation of human blood activity. It was observed that the extract showed greater percentage of inhibition of BSA (P potential platelet aggregation inhibitory activity in adose-dependent manner. The maximum inhibition was observed at the dose 400 μg/ml

Chilean crude extract of <i>Ruta graveolens</i>generates vasodilatation in rat aorta at subtoxic cellular concentrations

In Chile elevated percentage of population have cardiovascular diseases, 70% of this populations is a hypertensive peoples. The Ruta graveolens (Ruta) is a medicinal plant used in different parts of the world with different therapeutics properties like dermatologic as far as anti-helmintic properties. We analyzed the vascular action measuring the tension to identify vasodilator effect of Ruta extract in norm-tense rat’s aorta incubated and measured in isolated organ bath, and evaluating the toxicity effect in CRL-1730 cell line, through enzymatic assay (MTT), confocal microscopy (propidium iodide stain) and flow cytometry (TUNEL assay), including extracellular reactive oxygen species (ROS) production through luminescence assay. The results show with DE50 29 ± 0.1 μg/mL evidenced vasodilatation, partially endothelium-depend. The cytotoxicity showed with DE50 304.6 ± 2 μg/mL in enzymatic assay (MTT) while evidenced membrane permeability in high concentrations (1500 μg/mL), DNA fragmentation in absence of oxidative stress in only observed when high concentrations of Ruta are used over the cell culture. The vasodilatation activity is executed in subtoxic concentration and partially endothelium-depend without permeability effect in the membrane and deterioration of the cells viability suggesting a complex effect of Ruta preparation in the regulation of vascular tone.

Oxidative Stress Evaluation in Cancer-Induced Mice Treated with Ruta graveolens L. Stem lectins

The production of free radicals is a natural process in aerobic organisms due to the mitochondrial activity. Usually the cells have several mechanisms for scavenging those free radicals, but, in several pathologic processes, such as cancer, those free radicals increase their production, making it impossible to sustain the system stable, generating the condition called oxidative stress. Ruta graveolens L. （Rue） is a plant commonly used in traditional medicine, mainly as antiinflamatory; this has been related to some organic components, such as Rutin, but there hasn＇t been any lectin studies in Rue stem. Lectins are glycoproteins of non-enzymatic and no immune origin, able to bind to simple carbohydrates, which lets them bind selectively to malignant cells against normal cells, killing them via apoptosis and reducing the free radicals level. In this study we intended to characterize rue stem lectins as those weren＇t reported yet. Also, the anticancer and antioxidant activity of these lectins was evaluated, Rue stem lectins were extracted using a saline solution and semipurified to obtain an enriched extract and administrated to nickel oxide treated mice. Oxidative stress was cuantified using the tiobarbituric acid （TBARS） method to quantify Malondialdehyde （MDA）, the Griess method to cuantify Nitrites and enzymatic activity of catalase were cuantified in liver. In this study was found that rue stem lectins are useful as a therapeutic auxiliar, considering that its ratio of antioxidant activity is limited, being a prooxidant agent at high concentrations.

气相色谱-离子迁移谱法分析5种外观易混淆香辛料挥发性成分差异

目的比较5种外观易混淆香辛料挥发性成分差异。方法采用气相色谱-离子迁移谱法(gas chromatography-ion mobility spectroscopy,GC-IMS)分析孜然、小茴香、莳萝籽、百里香和千里香5种外观易混淆香辛料的挥发性成分,并通过指纹图谱和主成分分析法(principal component analysis,PCA)进行差异比较。结果5种香辛料样品中共鉴定出80种挥发性成分,且不同样品间挥发性成分种类及相对含量差异明显。枯茗醛、β-蒎烯等为孜然中特征挥发性成分,赋予其辛辣草药香、孜然香特征香气;香芹酮、柠檬烯等为莳萝籽中特征挥发性成分,赋予其辛香甘甜、温和而不刺激的特征香气;β-石竹烯、香茅醇等为千里香中特征挥发性成分,能够增强其特征香气的通透性;草蒿脑、罗勒烯、1,8-桉叶素、γ-松油烯、α-蒎烯、水芹烯、月桂烯等烯类,二丁基硫醚及乙酸芳香酯等酯类为小茴香中特征挥发性成分,共同赋予其强烈而偏甜润的特征香气;百里香酚、2-茨醇、正丁醇、芳樟醇、糠醛和2-乙酰基呋喃等为百里香中特征香气,共同赋予其清甜的药草香气。结论根据挥发性成分差异,可将5种外观易混淆香辛料进行有效区分。该研究为GC-IMS技术应用于香辛料的快速鉴别提供了理论参考。

臭草二氯甲烷部位化学成分研究

为研究臭草(Ruta graveolens L.)地上部分的化学成分及其抗Epstein-Barr病毒(EBV)活性。本研究采用硅胶等柱色谱和制备液相色谱等手段对臭草地上部分的95%乙醇浸提物二氯甲烷萃取部位进行分离纯化,并根据理化性质和谱学数据对化合物结构进行鉴定,鉴定后化合物进行抗EBV活性测试。从臭草乙醇提取物二氯甲烷萃取部分离得到30个化合物,分别鉴定为山小柑碱(1)、2-羟基-3-异丙氧基-1,4-甲氧基-10-甲基吖啶酮(2)、芸香日酮(3)、1-羟基-3-甲氧基-N-甲基吖啶酮(4)、羟甲基吖啶酮环氧化物(5)、rutalinium(6)、(-)-日把里尼定(7)、ribalinium(8)、(+)-ribaline(9)、γ-fagarine(10)、香草木宁碱(11)、合帕洛平(12)、茵芋碱(13)、(-)-去乙酰基芸香苦素(14)、chalepensin(15)、(+)-芸香瑞亭(16)、花椒毒素(17)、补骨酯素(18)、佛手柑内酯(19)、7-preniloxicumarin(20)、异东莨菪素(21)、6-羟基香豆素(22)、东莨菪苷(23)、芸香灵(24)、芸香宁(25)、3-(3,4,5-三甲氧基苯基)-2-丙烯-1-醇(26)、moskachan D(27)、胡椒基丙酮(28)、芦丁(29)、1,3-disinapoyl-gentiobiose(30),其中化合物2、7、9、12、20、22、23、26、27、28、30为首次从该植物中获得。将30个化合物均测试其体外抗EBV活性,结果显示在30μmol/L浓度下,共有6个化合物抑制EB病毒裂解复制达到50%以上,其中化合物5、24,显示较好的抗EB病毒活性,IC 50分别为2.4μmol/L与4.8μmol/L,高于阳性对照组(右旋芸香苦素,IC 50=7.0μmol/L)。

基于高效液相色谱法的芹菜11种酚类化合物分析及应用

以10种芹菜(Apium graveolens L.)为试材,采用Venusil XBP Cl8(L)色谱柱(250 mm×4.6 mm,5μm),以0.2%乙酸溶液-甲醇为流动相,梯度洗脱,流速为0.5 mL/min;采用PDA检测器,检测波长为240 nm。建立了基于高效液相色谱(HPLC)同时测定芹菜中11种酚类化合物的方法,并对色谱柱的选择、流动相条件和梯度洗脱条件等进行优化。结果表明,11种酚类物质在0.5~10.0μg/mL范围内线性关系良好,相关系数均≥0.990。在芹菜样品中进行加标水平为0.2、0.4、1.0 mg/kg的回收试验,回收率为71.90%~134.00%,相对标准偏差(RSD)为0.30%~4.00%。检出限(LOD)与定量限(LOQ)分别为0.07~0.29 mg/kg、0.20~0.84 mg/kg。各活性成分在24 h内稳定性良好,相对标准偏差均小于3.00%。该方法成功应用于10种芹菜中11种酚类化合物含量的测定,结合聚类分析发现不同品种的芹菜酚类组分存在相似性,但其含量存在品种间差异。该方法灵敏度高、精密度好,可为芹菜中11种酚类化合物含量的分析提供技术支持。

Discovery of antitumor diterpenoids from Casearia graveolens targeting VEGFR-2 to inhibit angiogenesis

Eight novel clerodane diterpenoids(1-8)were isolated from the twigs of Casearia graveolens.Their structures were elucidated through comprehensive nuclear magnetic resonance(NMR),high-resolution electrospray ionization mass spectrometry(HRESI-MS),and electronic circular dichroism(ECD)analyses.In addition to structural determination,surface plasmon resonance(SPR)assays were conducted to investigate molecular interactions,revealing that compound 8 exhibited high affinity for vascular endothelial growth factor receptor 2(VEGFR2),a key regulator of tumor angiogenesis.Subsequent in vivo experiments demonstrated that compound 8 effectively inhibited angiogenesis and displayed significant antitumor activity by suppressing tumor proliferation and metastasis in zebrafish xenograft models.These findings suggest that compound 8 holds promise as an anticancer lead compound targeting VEGFR-2 to obstruct tumor angiogenesis.

芹菜APX基因家族鉴定及其表达分析

抗坏血酸过氧化物酶(Ascorbate peroxidase, APX)是植物活性氧代谢中重要的抗氧化酶之一,在植物抵抗氧化胁迫方面发挥重要作用。利用生物信息学方法对芹菜基因组中的APX基因家族成员进行鉴定和分析,并通过实时荧光定量PCR(quantitative real-time PCR, qRT-PCR)验证分析AgAPXs在高温胁迫下的表达情况,为开展芹菜APX基因参与高温胁迫调控机制提供依据。结果表明:芹菜基因组中共有9个APX基因,随机分布在5个染色体上,并出现了基因片段复制现象;大多数基因被定位在细胞质中。系统发育分析表明,AgAPX基因家族可分为3个亚族,同一亚族中的成员具有相似的基因结构和基序。启动子顺式元件分析表明,大多数AgAPX基因含有多种与生长发育、植物激素和逆境胁迫相关的顺式元件。高温胁迫下,芹菜APX活性提高。qRT-PCR分析表明,AgAPXs在不同时间的高温处理下表达具有显著差异,并与转录组表达丰度相一致,AgAPX2、AgAPX3、AgAPX4、AgAPX5、AgAPX7的表达量和APX活性具有显著相关性,推测AgAPXs可能参与了芹菜抵御高温的调控过程。本研究初步鉴定并提供了芹菜APX基因家族成员信息,为今后进一步探索芹菜APX基因功能提供了重要的研究基础。

几种植物浮床的水质净化及水华防治效率

分别构建香根草Vetiveria zizanioides、黄菖蒲Iris pseudacorus、西芹Apium graveolens浮床及其不同组合的植物浮床,并在水体中加入6种不同种类的浮游植物及一种水生动物-斑马鱼Danio rerio,研究不同浮床系统对水质的净化效应及其对水生生物的影响。结果显示三种植物在浮床上能健康生长,其生物量、株高和根长均明显增加,且香根草和黄菖蒲的长势优于西芹。植物浮床均能有效防止水体的中化学需氧量(CODMn)的上升,但各生态浮床植物之间无显著性差异;植物浮床能显著去除水体中的氮(N)、磷(P)营养盐,其中,香根草的去除效果最好,而西芹的效果相对较差。总体而言,在系统中添加斑马鱼能在一定程度上提高浮床系统对水体的净化功能。香根草浮床能显著抑制水体中浮游植物的生长,相对而言其他类型的植物浮床对浮游植物的抑制能力较弱或无显著影响,且不同种类浮游藻类对植物浮床的敏感度不同。香根草和黄菖蒲对生态浮床系统中斑马鱼的生物量具有促进作用,香根草的促进作用大于黄菖蒲。综上所述,香根草是一种优良的浮床植物,可广泛用于水体净化及水华防治,而黄菖蒲可用于城市景观水体的净化。

基于RAPD分子标记芹菜品种亲缘关系分析研究

[目的]研究RAPD分子标记芹菜品种亲缘关系分析方法。[方法]以来自中国不同地区40种不同品种的芹菜为试材,对RAPD技术在鉴定芹菜品种中的科学性进行验证与分析。[结果]13种不同长度引物扩增的不同品种芹菜DNA的谱带清晰度以及数量等有明显区别,在相似系数为0.61时,可将所有测试芹菜品种划分为4类。[结论]RAPD技术是适用于分析芹菜品种亲缘关系的简易与理想的DNA分子标记技术,能很好的区分芹菜品种间亲缘关系的远近。

野生水芹与旱芹的营养成分比较分析

为研究野生水芹的营养价值,以湖北野生水芹和旱芹为材料,对新鲜叶片和叶柄的主要营养成分进行了比较分析。结果表明:两种材料叶片和叶柄的淀粉、粗纤维和维生素B1含量的差异不明显,游离氨基酸和维生素B2含量比旱芹低,但野生水芹含有丰富的蛋白质和钙,其叶柄中可溶性糖和磷的含量分别达29.70 mg/g和0.77mg/g,分别比旱芹高57.23%和37.50%;其叶片维生素C、铁含量分别达0.19 mg/g和0.35 mg/g,比旱芹分别高137.50%和191.67%,整株黄酮类物质含量是旱芹含量的2倍。这说明野生水芹具有较高的食用价值,是一种适合作为涝渍地区保护性开发的野生蔬菜资源。

芹菜水焯工艺的优化

以芹菜(Apium graveolens)为研究对象,对自来水、1%乙酸溶液、10 g/L食盐溶液和10 g/L小苏打溶液4种水焯介质进行了研究;在单因素试验的基础上,通过L9(34)正交设计探讨了芹菜水焯的工艺条件。结果表明,乙酸溶液可作为芹菜的水焯介质;芹菜水焯的最佳条件为乙酸体积分数0.02%,料液比1∶4(m/V,g∶mL),在100℃水焯3 min。在此条件下,水焯芹菜的干物质溶出率为0.41%。水焯后芹菜可用于加工各种菜肴。