Biology of ginseng (panax quinquefolium in Illinois /

On cover: Biology of ginseng in Illinois.

Investigating an Alternative Treatment to Cystic Fibrosis: Testing the Effect of Panax ginseng C.A. Meyer extracts on Pseudomonas aeruginosa

The predominant pathogen found in the lungs of cystic fibrosis (CF) patients is Pseudomonas aeruginosa. The success of the infection is partially due to virulence factor production, which is regulated by quorum sensing (QS) signaling. Currently, antibiotics are used to treat the infection, but resistant forms of P. aeruginosa have evolved, necessitating alternative treatments. Previous animal studies showed that treatment with extracts from the Chinese herb Panax ginseng C.A. Meyer reduced bacterial load resulting in a favorable immune response. It is hypothesized that ginsenosides, the major bioactive compounds in ginseng, is responsible for this effect. This study explores the role of ginseng extracts in attenuating P. aeruginosa virulence. A sequential extraction was performed using hexane, methylene chloride, methanol, and water. High performance liquid chromatography (HPLC) analysis showed the methanol and water ginseng extracts contained the known ginsenosides Rb1, Rb2, Rc, Rd, Re, and Rg1• All extracts were tested on biomonitor strains of Agrobacterium tumefaciens,Chromobacterium violaceum, and P. aeruginosa. Antibacterial and anti-QS activity were assessed using a disc diffusion assay. This was then followed by thin layer chromatography (TLC) bioautographic assay to further separate active compounds. The hexane and dichloromethane extracts, that lacked ginsenosides, displayed antibacterial activity against C. violaceum, whereas methanol and water extracts had anti-QS activity. The results of the bioassay with the pure ginsenoside standards showed that they lack antibacterial or anti-QS activity. Our results indicate that there are bioactive compounds, other than ginsenosides, that are the cause of antibacterial effects and anti-QS in the ginseng extracts.

Testing the Efficacy of Panax ginseng Extract as a Novel Anti-Quorum Sensing Agent

Quorum sensing (QS) is the phenomenon by which microorganisms regulate gene expression in response to cell-population density. These microorganisms synthesize and secrete small molecules known as autoinducers that increase in concentration as a function of cell density. Once the cell detects the minimal threshold concentration of an autoinducer, the gene expression is altered accordingly. Although the cellular circuitry responding to QS is relatively conserved, the cellular processes regulated by QS, for example are conjugation, motility, sporulation, biofilm formation and production of virulence factors importamt for infection. Since many pathogens have developed resistance to available antibiotics, novel therapies are required to further treat these pathogens. Inhibitors of QS are potential therapeutic candidates since they would inhibit virulence without selecting for antibiotic resistant strains. Previous studies have shown that the Chinese herb Panax ginseng contains compounds that inhibit QS activity. To further characterize this activity, a highly sensitive quantitative liquid assay was developed using a Chromobacterium violaceum AHL mutant, CV026 as a biomonitor strain. After confirmation that P. ginseng aqueous extracts had anti-QS activity, the extract was fractionated on a reverse phase C18 Sep Pak column. Most anti-QS activity was present in the flow through, and compounds eluted with ten percent acetonitrile in water antibacterial activity. We thus conclude that P. ginseng has anti-QS activity and the active compound is water soluble. Compounds from P. ginseng, could be used as a lead structure to design compound with higher anti-QS activity for therapeutic treatments.

西洋参体细胞胚胎发生的研究

以西洋参(Panax quinquefolium L.)为材料，从胚、胚乳、多年生主根等外植体诱导筛选得到多种胚性及非胚性愈伤组织，胚来源的胚性愈伤组织经用附加1.0ppm 2,4-D的MS培养基继代保持三年后仍具旺盛的体细胞胚胎发生能力，以胚性愈伤组织建立的液体培养系统可得到大量游离的胚状体，将胚状体包埋制成的人工种子能在附加0.1ppm NAA和1.0ppm GA_3的B_s培养基上萌发形成根、茎、叶健全的再生植株，并且移栽成活。松软型根愈伤组织以过长期继代培养后，也得到了胚状体发生。比较了多种培养因素对体细胞胚胎发生和愈伤组织的影响，其中外植体来源、基本培养基的无机盐组成以及生长素是决定愈伤组织形态结构类型和体细胞胚胎发生能力的主要因素。胚乳愈伤组织、质密型根愈伤组织以及来源于胚的松软型非胚性愈伤组织在多种诱导条件下均未能发生胚状体。 从松软型的胚或根愈伤组织均容易游离大量原生质体，用悬浮培养物游离原生质体的得率更高，从早期胚状体也可酶解得到可用于培养量的原生质体。原生质体体积均很小，培养中的行为也相似，有些形成细胞壁，细胞变形，个别进行细胞分裂，形成少数细胞团，但未形成愈伤组织;有些原生质体仍保持球形，体积剧裂膨大几十倍，小液泡汇聚成大液泡，有些液泡中积累紫红色素。 用石蜡切片、半薄切片、透射电镜和扫描电镜等方法对各种愈伤组织的内部结构及外部形态进行了详细观察，发现胚状体起源于愈伤组织的表皮或表皮下层细胞，有单个发生和成丛发生两种发生方式，不同愈伤组织细胞的显微及超微结构各具特色。胚乳愈伤组织、松软型根愈作组织以及胚状体的细胞虽然都具有某些胚性细胞结构特征，例如细胞体积小、细胞核大、细胞质浓厚、细胞器丰富等，但它们并不具有现实的体细胞胚胎发生能力。发生胚状体的细胞含大量多核糖体和内质网片段，小液泡数量少且形成多泡复合体，细胞壁也由于在组织中所外的位置不同而有厚薄两种类型之分，细胞常形成胚性细胞复合体，位于愈伤组织的外围，边缘可分化为许多小细胞团。胚性细胞复合体的表面质密平整，有许多丝状物和颗粒，细胞轮廓不清，但有显著突出的小细胞，细胞表面具沟脊;而其它各型愈伤组织的表面均为球形或半球形的大型细胞，表面仅具细小的纹理、凸起或颗粒。 胚来源的胚性愈伤组织的总蛋白组成与来源相同，但形态结构截然不同的非胚性愈作组织的总蛋白组成差异显著，后者与形态结构特征相似的根本源公软型愈伤组织具有相似的蛋白质电泳带型，但多一条33KD的蛋白带;培养基中去掉2,4-D后，松软型根愈伤组织的蛋白质组成发生轻微变化。等电聚焦和SDS聚丙烯酰胺凝胶双向电泳揭示出在单向电泳扫描上呈现最高峰的17KD蛋白质在胚性愈伤组织中随等电点的不同而分离，在其它三种构软型愈伤组织中则仅集中在等电点为5.9处，为一个大而染色深的点。 同样培养基上培养的形态结构不同的愈伤组织所含元素的量不同，说明细胞对无机盐的吸收是有选择性的，不同类型的愈伤组织可积累不同的元素。培养基中的元素组成情况在愈伤组织中也有一定反映，在含钠、钾元素较多的B_5培养基上生长的愈伤组织中这两种元素的含量也较高，培养基的离子胁迫作用和细胞对离子一定程度的被动吸收会影响细胞的代谢方式，从而导致细胞类型的分化，基本培养基对愈伤组织类型的转变及体细胞胚胎发生能力的影响可能正是通过影响细胞的离子吸收、代谢平衡而实现。

三七总皂苷对海马CA1区长时程增强效应的影响

三七总皂苷(Panax notoginseng saponins,PNS)是从传统中药三七的根中提取的主要有效成分,具有改善血液循环、耐缺氧、改善记忆力、抗衰老等多方面的生理活性.本研究采用"盲法"全细胞膜片钳技术观察PNS对大鼠海马CA1区锥体神经元长时程增强效应(LTP)的影响,以分析其增强学习记忆功能的神经电生理机制.以断头法分离Wistar大鼠(3～4 周)海马半脑,用切片机切出400 μm厚度的海马脑片,以全细胞电压钳制方式记录CA1区锥体细胞的兴奋性突触后电流(EPSCs),给予高频刺激HFS(100 Hz)诱导LTP,分析PNS对大鼠海马CA1区EPSCs和LTP的影响.结果表明,PNS(0.1～0.4 g·L-1)能显著抑制EPSCs(P＜0.05),且对海马CA1区LTP无易化作用;但PNS(0.04～0.05 g·L-1)不影响CA1区的EPSCs基础突触传递(P＞0.05),却可以增强HFS诱发的LTP(P＜0.05).上述结果提示,PNS(0.04～0.05 g·L-1)能易化海马CA1区锥体神经元的长时程增强效应,该作用应是其增进学习记忆力的神经电生理机制.

三七总皂苷对大鼠海马CA1区突触传递的作用及机制

下载PDF阅读器目的 研究三七总皂苷(Panax notoginseng saponins,PNS)对大鼠海马脑片CA1区锥体神经元兴奋性和抑制性突触传递的作用.方法 断头法分离3～4周雄性Wistar大鼠海马半脑,用切片机切出400μm厚度的海马脑片,对CA1区锥体细胞采用"盲法"全细胞膜片钳技术记录,分别检测和分析PNS(0.05～0.4 g/L)对刺激CA1传人纤维引出的兴奋性突触后电流(EPSCs)和抑制性突触后电流(IPSCs)的影响,继而以脉冲间隔为50 ms的配对刺激代替单刺激,通过EPSC2/EPSC1(P2/P1)值的变化观察PNS对双脉冲易化(paired-pulse facilitation,PPF)的影响.结果 0.1～0.4 g/L PNS显著抑制EPSCs(P＜0.05),且PNS在抑制P1、P2的同时明显升高P2/P1值(P＜0.05),加强了双脉冲易化,但PNS对IPSCs无显著影响(P＞0.05).结论 PNS 显著减小大鼠海马CA1区锥体神经元的EPSCs而不影响IPSCs,说明PNS不是通过强化抑制性中间神经元的功能间接地抑制兴奋性神经元,而是对兴奋性突触传递直接产生抑制;PNS明显升高P2/p1值,说明 PNS是通过突触前机制抑制CA1区兴奋性突触传递.

三七总皂苷对大鼠海马CA1区兴奋性突触活动的影响

目的:研究三七总皂苷(Panax notoginseng saponins,PNS)对海马脑片CA1区锥体神经元兴奋性突触活动的作用和机理。方法:采用"盲法"全细胞膜片钳技术,记录PNS(0.05~0.4 g/L)对3~4周雄性wistar大鼠海马脑片(400μm)CA1区兴奋性突触后电流(excitatory post synaptic currents,EPSCs)和自发的微小兴奋性突触后电流(miniature excitatory post synapticcurrents,mEPSCs)幅度及频率的影响。结果:0.1~0.4 g/L PNS显著抑制海马脑片CA1区EPSCs(P<0.05);0.05~0.4 g/LPNS可明显增加CA1区锥体神经元自发mEPSCs的产生频率,但并不影响mEPSCs的幅度。结论:PNS可作用于突触前位点对海马神经元兴奋性突触活动产生调节作用,PNS增加mEPSCs频率的作用可能与促进突触前膜nAChR的激动有关,这可能是其调节海马神经元的兴奋性进而发挥益智作用的机制之一;PNS对EPSCs和mEPSCs的不同作用说明PNS是选择性抑制膜去极化所诱发的递质释放过程,PN...

Effects of ultrahigh pressure extraction conditions on yields and antioxidant activity of ginsenoside from ginseng

Ultrahigh pressure technique was employed to extract ginsenosides from roots of ginseng (Panax ginseng C.A. Meyer). The optimal conditions for ultrahigh pressure extraction (UPE) of total ginsenosides were quantified by UV-vis spectrophotometry with the ginsenoside Re as standard, the signal ginsenosides were quantified by HPLC and ELSD with ginsenosides Re, Rg(1), Rb-1, Rc and Rb-2 as standards. Orthogonal design was applied to evaluate the effects of four independent factors (extraction pressure, extraction temperature, extraction time and ethanol concentration) on the yield and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of ginsenoside, which are based on microwave extraction (ME), ultrasound extraction (UE), soxhlet extraction (SE) and heat reflux extraction (HRE) method. The results showed that UPE method can produce ginsenoside with the highest yield and the best radical scavenging activity compared to other used ones. Scanning electron microscopic (SEM) images of the plant cells after ultrahigh pressure treatment was obtained to provide visual evidence of the disruption effect.

Investigation of the hydrolysis of ginsenosides by high performance liquid chromatography-electrospray ionization mass spectrometry

The hydrolysis of ginsenoside standards and the crude extracts of ginseng has been investigated at different pH values (2.4 - 11.2) using high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS). The experimental results indicated that the pH value of aqueous solutions is an important factor in changing the composition of ginsenosides. For (20S)-protopanaxadiol ginsenosides, ginsenosides with a large mass hydrolyzed to form hydrolysates (20S)-Rg(3) and (20R)-Rg(3) at pH 4.3. There were more hydrolyzed products observed at pH 3.3: (20S)-F-2, C-25,26 hydrated ginsenoside "C-Y-1" and "C-Y-2" (MW = 802 Da) accompanied with (20S)-Rg(3), (20R)-Rg(3). At pH 2.4, only (20R)-Rg(3), (20S)-F-2, a small quantity of (20S)-Rg(3) and three C-25,26 hydrated ginsenosides were obtained. For (20S)protopanaxatriol Re, no hydrolysates were observed at pH 4.3; it was hydrolyzed at pH 3.3 to form hydrolysates (20S)-Rg, (20R)Rg(2) and hydrated C-25,26 (MW = 802 Da) and at pH 2.4 only C-25,26 hydrated ginsenosides "C-Y-1" and "C-Y-2" (MW = 802 Da) were left in the solution. Similar hydrolysis reactions could be also observed for the crude extracts of ginseng. It showed that HPLC/ESI-MS is a fast and convenient method to study the hydrolysis of ginseng.

Studies on the content variation of chemical constituents during the combination of ginseng with veratrum nigrum by ESI-MS and HPLC-ESI-MS

To study the content variation of ginsenosides and alkaloids during combination of ginseng with veratrum nigrum, the ginsenosides and alkaloids in the decoction of ginseng with veratrum nigrum were analyzed and compared by high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) and electrospray ionization-mass spectrometry (ESI-MS). In the compatible decoction, eight ginsenosides and eight alkaloids. were detected, and the contents of six ginsenosides were found to be reduced, on the contrary, the contents of six alkaloids were increased. During combination of ginseng with veratrum nigrum, the contents of ginsenosides were reduced and those of the toxic alkaloids were increased. From the chemical point of view, the traditional theory is right that ginseng and veratrum nigrum are incompatible with each other.

Structural analysis of saponins from medicinal herbs using electrospray ionization tandem mass spectrometry

The underivatized saponins from Tribulus terrestris and Panax ginseng have been investigated by electrospray ionization multi-stage tandem mass spectrometry (ESI-MSn). In ESI-MS spectra, a predominant [M + Na](+) ion in positive mode and [M - H](-) ion in negative mode were observed for molecular mass information. Multi-stage tandem mass spectrometry of the molecular ions was used for detailed structural analysis. Fragment ions from glycoside cleavage can provide information on the mass of aglycone and the primary sequence and branching of oligosaccharide chains in terms of classes of monosaccharides. Fragment ions from cross-ring cleavages of sugar residues can give some information about the linkages between sugar residues. It was found that different alkali metal-cationized adducts with saponins have different degrees of fragmentation, which may originate from the different affinity of a saponin with each alkali metal in the gas phase. ESI-MSn has been proven to be an effective tool for rapid determination of native saponins in extract mixtures, thus avoiding tedious derivatization and separation steps.

Synthesis and evaluation of novel polysaccharide-Gd-DTPA compounds as contrast agent for MRI

Macromolecular conjugates of two kinds of natural polysaccharides, that from Panax quinquefolium linn (PQPS) and Ganoderma applanatum pat (GAPS), with gadolinium-diethylenetriaminepenta-acetic acid (Gd-DTPA) have been synthesized and characterized by means of FTIR, elementary analysis and ICP-AES. Their stability was investigated by competition study with Ca2+, EDTA (ethylenediaminetetraacetic acid) and DTPA. Polysaccharide-bound complexes exhibit T-1 relaxivities of 1.5-1.7 times that of Gd-DTPA in D2O at 25degreesC and 9.4T. MR imaging of Sprague-Dawley (SD) rats showed remarkable enhancement in rat liver and kidney after i.v. injection of these two complexes: liver parenchyma 60.9+/-5.6%, 57.8+/-7.4% at 65-85 min; kidney 144.9+/-14.5%, 199.9+/-25.4% at 10-30 min for PQPS-GdDTPA, GAPS-Gd-DTPA at gadolinium dose of 0.083 and 0.082 mmol/kg, respectively. Our preliminary in vivo and in vitro study indicates that the two kinds of polysaccharide-bound complexes are potential tissue-specific contrast agents for MRI.

Rapid identification of saponins in plant extracts by electrospray ionization multi-stage tandem mass spectrometry and liquid chromatography/tandem mass spectrometry

Electrospray ionization multi-stage tandem mass spectrometry (ESI-MSn) and liquid chromatography coupled with on-line mass spectrometry (LC/MS/MS) were applied to characterize saponins in crude extracts from Panax ginseng. The MSn data of the [M - H](-) ions of saponins can provide structural information on the sugar sequences of the saccharide chains and on the sapogins of saponins. By ESI-MSn, non-isomeric saponins and isomeric saponins with different aglycones can be determined rapidly in plant extracts. LC/MS/MS is a good complementary analytical tool for determination of isomeric saponins. These approaches constitute powerful analytical tools far rapid screening and structural assignment of saponins in plant extracts. Copyright (C) 2000 John Wiley & Sons, Ltd.

Studies on negative electrospray ionization mass spectrometry on saponins

Two kinds of saponins have been investigated by negative electrospray ionization (NESI) mass spectrometry. Under ESI conditions, the [M - H](-) ions of saponins were observed which provide the molecular weights of saponins. The fragment pathways of [M - H](-) ions of these two saponins depend on their structures. For steroidic saponins, [M - H](-) ion only produces the fragment ions by the losses of sugar units. For oleanolic saponins, [M - H](-) ion yields the cross-ring ions as well as the fragment ions by the losses of sugar units. Moreover, the abundance of the former is higher than that of the latter. The characteristic fragments are used to provide the sequence and some linkage information of sugar moieties of saponins. Especially, their fragment difference strongly depends on the linkage between the aglycone and the sugar moieties.

中国红参中碳、氢、氮元素和蛋白含量的自动分析

人参为五加科植物人参(Panax ginseng C.A.Mey.)的干燥根。红参则是鲜人参洗净后经高温蒸气加热后干燥而制得的产品。有关人参中皂甙、氨基酸、多肽、挥发油、微量元素等成分的研究国内外均有大量的报道。但关于红参中碳、氢、氮元素的研究以及自动定量方法国内外尚未见报道。 1.仪器:1106型元素自动分析仪(意大利)其配套设备包括B-5000型记录仪,微处理机和电子天平。 2.人参样品:1号红参由吉林省抚松县参茸公司提供。2号红参由抚松县漫江参场提供。3号红参为抚松县参场提供。4号红参由延边地区提供。5号红参由集安县新开河参场提供。上述样品均为6年生。