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.

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

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。

HPLC法同时测定旱芹中木犀草素和旱芹素的含量

建立HPLC法同时测定旱芹中木犀草素和旱芹素含量的方法。采用C1(8 150 mm×4.6 mm,5μm)柱,流动相为乙腈∶0.01 mol/L醋酸铵(pH=4.8)=40∶60,等强度洗脱,检测波长330 nm,柱温25℃,流速1.0 mL/min。木犀草素和旱芹素的质量浓度分别在0.93μg/mL^4.94μg/mL(r=0.999 1)及1.03μg/mL^5.06μg/mL(r=0.999 4)范围内与峰面积呈现良好的线性关系,平均加样回收率依次为99.7%(RSD为2.4%),99.0%(RSD为1.8%)。

芸香药材中花椒毒素的HPLC测定方法研究

目的:筛选出在芸香中提取花椒毒素的方法,建立HPLC法测定花椒毒素的含量。方法:在避光条件下,利用花椒毒素在二氯甲烷和乙酸溶液中均有较好溶解度,通过二氯甲烷提取,挥干,残渣加5%乙酸溶液溶解,过滤,进样。采用C_(18)色谱柱,流动相为乙腈:0.02 mol·L^(-1)乙酸铵(45∶55),流速1.0 mL·min^(-1),检测波长300 nm,柱温35℃。结果:花椒毒素在0.804~80.4μg·mL^(-1)范围内线性关系良好(r=0.9999);平均加样回收率为96.4%,RSD=4.0%。不同产地的10批样品测定结果为:0.04%~0.31%,RSD(n=3)为0.85%~1.96%。结论:本实验建立的芸香中提取花椒毒素的方法及HPLC法能有效地测定芸香中花椒毒素的含量,本法已列入芸香药材质量标准(草案)。

Differences of cadmium absorption and accumulation in selected vegetable crops

A pot experiment and a sandy culture experiment grown with three vegetable crops of Chinese cabbage ( B. chinensis L., cv. Zao\|Shu 5), winter greens ( B.var.rosularis Tsen et Lee, cv. Shang\|Hai\|Qing) and celery ( A.graveolens L. var. dulce DC., cv. Qing\|Qin) were conducted, respectively. The initial soil and four incubated soils with different extractable Cd (0.15, 0.89, 1.38, 1.84 and 2.30 mg Cd/kg soil) were used for the pot experiment. Five treatments were designed (0, 0.0625, 0.125, 0.250 and 0.500 mg Cd/L) in nutrient solution in the sandy culture experiment. Each treatment in pot and sandy culture experiments was trireplicated. The objectives of the study were to examine Cd accumulation in edible parts of selected vegetable crops, its correlation with Cd concentrations in vegetable garden soil or in nutrient solution, and evaluate the criteria of Cd pollution in vegetable garden soil and in nutrient solution based on the hygienic limit of Cd in vegetables. Cadmium concentrations in edible parts of the three selected vegetable crops were as follows: 0.01—0.15 mg/kg fresh weight for Chinese cabbage, 0.02—0.17 mg/kg fresh weight for winter greens, and 0.02—0.24 mg/kg fresh weight for celery in the pot experiment, and 0.1—0.4 mg/kg fresh weight for Chinese cabbage, 0.1—1.4 mg/kg fresh weight for winter greens, and 0.05—0.5 mg/kg fresh weight for celery in the pot experiment(except no\|Cd treatment). The order of the three test vegetable crops for cadmium accumulation in the edible parts was celery>winter greens>Chinese cabbage in both the pot experiment and the sandy culture experiment. Cadmium accumulation in edible parts or roots of the vegetable crops increased with increasing of cadmium concentration in the medium(soil or nutrient solution). And cadmium concentrations in edible parts of the test vegetable crops were significantly linearly related to the Cd levels in the growth media (soil and nutrient solution). Based on the regression equations established and the limit of cadmium concentration in vegetable products, the thresholds of Cd concentration in the growth medium evaluated was as follows: 0.5 mg/kg soil of extractable Cd for soil and 0.02 mg /L for nutrient solution. The high capacity for cadmium accumulation in the edible parts of different vegetable crops together with the absence of visual symptoms implies a potential danger for humans.

HPLC法测定芸香中4个成分的含量

目的测定芸香中芸香碱、花椒毒素、茵芋碱和白鲜碱4种成分的含量。方法采用HPLC法测定芸香中芸香碱、花椒毒碱、茵芋碱和白鲜碱4种成分的含量。色谱柱为Agilent Eclipse C18(250mm×4.6 mm,5μm);流动相为乙腈-0.02 mol/L甲酸铵溶液,梯度洗脱;流速为1.0 mL/min;检测波长为330 nm;柱温35℃。结果芸香碱、花椒毒素、茵芋碱和白鲜碱的线性范围分别为0.58~11.66μg/mL(r=0.9999)、1.12~22.40μg/mL(r=0.9999)、1.51~30.20μg/mL(r=0.9999)、1.14~22.84μg/mL(r=0.9997);平均回收率(n=6)分别为:100.71%(RSD=2.50%)、95.25%(RSD=3.38%)、106.67%(RSD=2.38%)、93.86%(RSD=0.88%)。6批次样品中上述4种成分的平均含量分别为0.1871、1.0812、0.3246、0.1473 mg/g。结论本方法操作简便,稳定性好,适用于芸香药材中主要成分的含量测定。

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)。

Evaluation of chemopreventive potentials of ethanolic extract of Ruta graveolens against A375 skin melanoma cells in vitro and induced skin cancer in mice in vivo

OBJECTIVE： Chemopreventive approach with natural products, particularly plants and plant-derived ones, is receiving increasing attention for their effective role against cancer without any palpable side effects. In this study, efficacy of ethanolic extract of Ruta graveolens （RG） on skin melanoma cells （A375） in vitro and on 7,12-dimethylbenz（a）anthracene （DMBA）-induced skin cancer in vivo has been tested in Swiss albino mice. METHODS： Studies on cell viability, apoptosis and autophagy induction were conducted in vitro. To check apoptosis, assays like alteration in mitochondrial membrane potential, annexin V-fluorescein isothiocyanate/propidium iodide assay and immunoblot were performed. Fluorescence microscopic and immunoblot assays were performed to confirm autophagy induction. The effects of RG were determined by evaluating body weight, tumor incidence, tumor volume and tumor burden in mice. Enzymatic and non-enzymatic antioxidant status was assessed. The role of some relevant signaling proteins was also analyzed. RESULTS： RG caused death ofA375 cells through induction of caspase 3-mediated apoptosis and Beclin-l-associated autophagy. Moreover, RG administration （75 mg/kg body weight） which showed no acute or chronic toxicity, showed significant reduction in the skin tumor burden of DMBA-painted mice. RG also demonstrated potent anti-lipid peroxidative and antioxidant functions during the course of skin cancer induction by DMBA. CONCLUSION： Chemopreventive potential of RG was demonstrated from overall results of this study indicating its possible use in therapeutic formulation of an effective drug to treat skin cancer.

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。

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。

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。

维药芹菜籽化学成分研究

从维吾尔药芹菜籽中分离得到4个化合物,经光谱鉴定为对羟基苯甲醛(1),3-羟基-4-异丙基苯甲酸(2),香草酸(3),4-羟基-2-异丙基-5-甲基苯基-1-O-β-D-葡萄糖苷(4)。4个化合物均为首次从该植物种子中分离得到。

芸香叶片和花瓣释放挥发性有机物成分及其变化规律

为探究芸香Ruta graveolens叶片和花瓣释放的挥发性有机物的成分及其变化规律,采用顶空-固相微萃取与气相色谱-质谱联用技术,对芸香叶片和花瓣中释放的挥发性有机物成分进行采集,分析叶片和花瓣中挥发性有机物的主要成分、相对含量及叶片在不同时期挥发性有机物成分的差异。结果表明:1 a中不同季节芸香释放的挥发性有机物共鉴定出29种,其中酯类化合物11种,酮类化合物7种,为芸香挥发物的主要成分。1 a中,芸香叶片释放的挥发性有机物的种类呈先增加后减少的趋势;其中,6月检测到的挥发性有机物种类最多,为18种;9月最少,仅为10种。主要成分酯类物质的相对含量呈先上升后下降趋势,酮类化合物呈先下降后上升趋势。在6月的日变化检测中,挥发性有机物的种类呈先增多后减少趋势,依次为12:00>10:00>14:00>16:00,18:00>8:00>20:00,在12:00达到高峰,以酯类和酮类化合物为主。芸香花瓣释放的挥发性有机物种类少于叶片,主要成分为酮类化合物,以2-十一(烷)酮为主。芸香叶片和花瓣释放的挥发性有机物的主要成分均具有一定的药用保健作用,因此,芸香是营造保健型植物景观的理想材料。

离子液体超声辅助提取香叶总黄酮及其抗氧化性研究

探索离子液体超声辅助提取香叶总黄酮的工艺条件及其体外抗氧化活性。以香叶总黄酮提取率为考察指标,采用单因素及正交试验对香叶总黄酮的提取工艺进行考察;应用紫外-分光光度法测定了香叶总黄酮对DPPH·的清除率。试验结果说明香叶总黄酮的最佳提取工艺条件为:提取溶剂为40%乙醇水溶液,料液比为1∶50、离子液体为氯化-1-丁基-3-甲基咪唑,离子液体浓度为0.35mol/L,提取水浴温度为65℃、提取时间为3h、超声波频率为90 W,在上述最佳提取工艺参数下,离子液体超声辅助提取香叶总黄酮的提取率可以达到109.79mg/g。体外清除自由基试验结果表明香叶总黄酮清除DPPH·的IC50为0.2360mg/mL,说明香叶总黄酮具有显著的体外抗氧化活性。研究结果为香叶的进一步开发利用提供了依据。