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.

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.

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.

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。

芹菜NHL-like蛋白基因克隆与表达分析

植物生长发育过程中会遭遇各种病原物的攻击,植物为了应对这些危害并适应外界的生存竞争,逐渐演化出了复杂的防御机制。NHL基因家族庞大,部分NHL基因受病原物诱导后会过量表达,增强植物对多种病原菌的抗性,是与植物防御机制密切相关的蛋白。本研究以津南实芹和美国西芹2个芹菜品种为试验材料,分别克隆出NHL-like蛋白基因AgNHL。序列分析表明,上述2种芹菜的AgNHL基因序列均含636 bp的开放阅读框,编码211个氨基酸。2种芹菜碱基序列,只有第36位不同,津南实芹为T,美国西芹为C,2种碱基序列相似性高达99.84%,编码的氨基酸序列相同。进化分析显示,2种芹菜的NHL-like蛋白与葡萄、大豆等植物的相似度较高,在第80～185氨基酸间含一个LEA-2蛋白保守结构域。荧光定量PCR结果表明,AgNHL基因主要在芹菜茎中表达,根中表达量最低,有明显组织特异性,品种差异也很显著。对2种芹菜分别进行4℃低温、38℃高温、20%PEG处理、0.2 mol/L NaCl处理2 h表达分析显示,低温、盐处理下该基因表达量明显上升。

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%)。

芹菜AgLIMS基因的克隆和生物信息学分析及其表达特性

以芹菜(Apium graveolens Linn.)品种‘六合黄心芹’(‘Liuhe Huangxinqin’)和‘津南实芹’(‘Jinnan Shiqin’)为研究对象,分别克隆获得芹菜柠檬烯合酶基因,命名为AgLIMS,其开放阅读框长1800 bp,编码599个氨基酸。结果显示:2个芹菜品种的AgLIMS基因核苷酸序列存在2个位点的差异,其编码的氨基酸序列也存在2个位点的差异;AgLIMS基因编码的氨基酸序列在第50至第586位存在植物萜类合酶(Terpene cyclase plant C1)保守区域,具有典型的类异戊二烯合成酶(Isoprenoid Biosyn C1)超家族结构域。2个芹菜品种的AgLIMS蛋白的理论相对分子质量分别为69120和69190,理论等电点均为pI 5.52。AgLIMS蛋白的氨基酸序列中,亲水性氨基酸比例高于疏水性氨基酸;其二级结构主要由α螺旋和无规则卷曲构成,但α螺旋、延伸链和β转角的比例在2个芹菜品种间略有差异;其三级结构均含有1个β折叠,但α螺旋数量在2个芹菜品种间略有差异。AgLIMS基因与多种植物LIMS基因编码的氨基酸序列一致性为61.33%;在系统树上,2个芹菜品种与同科种类的进化关系较近,而与其他科木本种类的进化关系较远。qRT-PCR结果显示:AgLIMS基因在2个芹菜品种的叶片、叶柄和根中均能表达,其相对表达量在叶片中最高、在根中最低,且在不同组织间差异显著;该基因在‘六合黄心芹’不同组织中的相对表达量总体上显著高于‘津南实芹’。在高温(38℃)、低温(4℃)、干旱(质量体积分数20%PEG6000)和高盐(0.2 mol·L^-1NaCl)条件下,2个芹菜品种AgLIMS基因的相对表达量呈现不同的变化趋势;总体上看,‘津南实芹’AgLIMS基因的相对表达量均不同程度升高,而品种‘六合黄心芹’AgLIMS基因的相对表达量则在高温和干旱条件下显著升高。综合分析结果表明:2个芹菜品种的AgLIMS基因核苷酸序列及其编码的氨基酸序列差异很小;AgLIMS蛋白为中性、亲水性蛋白,其进化具有一定的保守性;AgLIMS基因在芹菜中的表达具有组织特异性,且在受到非生物胁迫时其表达特性发生变化,但这种变化效应存在品种间差异。

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。

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。

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)。

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。

旱芹膳食纤维对高脂血症大鼠血脂的影响

目的建立高脂血症大鼠模型,研究旱芹膳食纤维(DF)对高脂血症大鼠各项血脂指标的影响。方法 Wistar大鼠随机分为正常对照组,高脂模型组,降脂灵对照组,旱芹DF低、中、高剂量组。以高脂饲料喂养大鼠10d建立高脂血症大鼠模型,正常对照组和高脂模型组均灌胃等量生理盐水,旱芹DF低、中、高剂量组和降脂灵对照组均灌胃给药,18d后称量体重,并用酶法测定血浆中胆固醇(TC)、甘油三酯(TG)和高密度脂蛋白胆固醇(HDL-C)的水平,评价旱芹DF的降血脂功能。结果以高脂饲料喂养大鼠10d后,与正常对照组相比,高脂模型组TC、TG和体重增量均升高且有显著性差异(P<0.05),从而成功建立了混合型高脂血症大鼠模型。以不同剂量旱芹DF混悬液灌胃18d后,各组血浆TC、TG水平与高脂模型组比较,具有显著性差异(P<0.05);旱芹DF低、中剂量组TC和动脉硬化指数(AI)水平降低明显,与降脂灵对照组比较,具有显著性差异(P<0.05);旱芹DF低剂量组TG水平与降脂灵对照组比较,具有显著性差异(P<0.05);旱芹DF高剂量组与低剂量组比较,TC、TG、AI水平具有显著性差异(P<0.05)。结论 DF的摄入量越大,其改善血脂水平的效果越好,显示出明显的剂量依赖关系,提示旱芹DF具有调节血脂的作用。

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。

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。