In Vivo radioprotective effect of Panax ginseng C.A. Meyer and identification of active ginsenosides

This study evaluated the effect of water extracts of Panax ginseng C.A. Meyer (PG), panaxadiol (PD), panaxatriol (PT), ginsenoside Rb(1), Rb(2), Rc, Rd, Re and Rg(1) on jejunal crypt survival, endogenous spleen colony formation and apoptosis in jejunal crypt cells in gamma-irradiated mice. Jejunal crypts were protected by pretreatment with PG, Rc and Rd. Administration of PG, PD, Rd and Re prior to irradiation resulted in an increase in the formation of endogenous spleen colonies. The frequency of radiation-induced apoptosis in intestinal crypt cells was also reduced by pretreatment with PG, PD, Rb(2), Rc, Rd, Re and Rg(1). In experiments on the effects of the individual ginsenosides, the rank order of activity was Rc > Rd > Rg(1) > Rb(2) > Re > Rb(1) on intestinal crypt survival assay, Re > Rb(2) > Rd > Rg(1) > Rb(1) > Rc on the spleen colony formation assay, and Rg(1) > Re > Rd > Rc > Rb(2) > Rb(1) on inhibiting the death of cells caused by apoptosis. The results indicated that Rc, Rd and Re may have a major radioprotective effect in mice irradiated with high and low doses of radiation. When the same experiments were performed using PD and PT, it was observed that most of the inhibitory effects came from PD rather than PT.     

西洋参不同器官中皂苷量与鲨烯合成酶和鲨烯环氧酶基因表达的相关性

目的探讨西洋参不同组织器官中总皂苷和单体皂苷量的差异及其与人参皂苷合成途径中鲨烯合成酶(SQS)和鲨烯环氧酶(SQE)基因表达量之间的关系。方法以4年生西洋参的14个组织器官为材料,索氏回流法提取总皂苷,采用HPLC法测定6种单体皂苷(人参皂苷Rg1、Re、Rb1、Rc、Rb2和Rd)的量,用香草醛-硫酸比色法测定总皂苷量。通过实时荧光定量PCR分析SQS和SQE基因在西洋参14个组织器官中的表达量。结果总皂苷和单体皂苷量在西洋参不同组织器官中差异非常显著(P<0.01)。除人参皂苷Rb2外,各单体皂苷及总皂苷之间均呈非常显著的正相关关系(P<0.01)。SQS和SQE基因在14个组织器官中的表达量具有非常显著的差异(P<0.01)并与人参皂苷Re、Rg1、Rb1、Rd和总皂苷量之间有显著的正相关关系(P<0.05)。结论西洋参不同组织器官中皂苷的量存在差异,SQS和SQE基因在人参皂苷合成途径中起着极其重要的作用。     

基于人参皂苷含量的西洋参等级规格标准研究

目的：分析不同产地、不同等级规格西洋参中人参皂苷的含量,评价西洋参的化学信息,为其行业标准的制定提供实验依据。方法：采用UPLC测定主要活性成分人参皂苷Rg1、Re、Rb1、Rb2、Rb3、Rc、Rd、Rg2含量,用SPSS16.0软件进行统计分析。结果：建立了不同产地西洋参样品人参皂苷分布特征图谱;西洋参中Rg1、Re、Rb1的含量之和符合《中国药典》标准;方差分析表明产地、长度对同一个规格的西洋参中人参皂苷的含量影响具有统计学意义（P〈0.01）。结论：由于西洋参中人参皂苷含量受产地、长度、重量等多因素影响,在等级规格评价中,符合《中国药典》标准基础上不宜采用人参皂苷含量为指标进行等级规格划分。     

基于孕烷受体-细胞色素 P450 3A4通路筛选复方丹参中的效应成分实验研究

目的采用孕烷受体-细胞色素 P450 3A4 （pregnane X receptor-cytochrome P450 3A4,PXR-CYP3A4）工程细胞株筛选复方丹参中诱导或抑制PXR-CYP3A4通路的化学成分。方法利用实验室构建的PXR-CYP3A4稳定转染人肝母细胞癌（human hepatoblastoma G2,HepG2）工程细胞株结合报告基因技术,筛选复方丹参中诱导或抑制PXR-CYP3A4通路的化学成分,并从酶活性水平进行确证。实验分为阳性对照组（RIF,10 μmol/L）、溶剂对照组（DMSO,0.1%）、药物处理各剂量组（人参皂苷Rc、Rf、Rb2、Rg2、F2、F1、Re,丹参酮Ⅰ,异龙脑5、10、25、50、100、200 μmol/L）,每种药物设置6个复孔,加药36、48 及60 h后吸取细胞培养液上清,测定CYP3A4活性,计算诱导倍数。结果与溶剂对照组比较,人参皂苷Rc、Rf、Rb2、Rg2、F2、F1、Re,丹参酮Ⅰ,异龙脑50、100 μmol/L处理30、48和60 h时,诱导倍数均升高（P〈0.05）,其中药物浓度200 μmol/L组处理48和60 h时,人参皂苷Rb2、Rg2、F1组诱导倍数均显著高于RIF阳性对照组（P〈0.05）。人参皂苷48 h时Rc、Rf、Rb2、F2、F1组CYP3A4酶活性提高（P〈0.05）。结论复方丹参中的人参皂苷Rc、Rf、Rb2、F2、 F1,丹参酮Ⅰ及异龙脑可以诱导CYP3A4酶。     

人参皂苷对DC2.4吞噬抗原和表达CD40分子的影响

目的:观察人参皂苷Rg1、Rb1及其联合使用对树突状细胞系DC2.4吞噬抗原和表面分子表达的影响。方法:体外培养DC2.4细胞,分别加入不同剂量人参皂苷Rg1、Rb1及Rg1+Rb1孵育24h后,分别加入异硫氰酸荧光素标记的卵白蛋白(FITC-OVA),流式细胞仪检测DC2.4细胞吞噬卵白蛋白抗原的情况;体外培养DC2.4细胞,分别加入LPS和不同剂量人参皂苷孵育24h,流式细胞仪检测细胞表面分子CD40的表达。结果:人参皂苷Rg1、Rb1、Rg1+Rb1在一定浓度可促进DC2.4细胞抗原吞噬的能力;三者均抑制CD40的表达。结论:人参皂苷Rg1、Rb1可通过影响DCs的抗原吞噬功能来发挥免疫调节作用;Rg1、Rb1联合使用与单独应用比较无明显差异。     

人参美白霜中人参皂苷的SPE-ESI-MS/MS分析

目的：应用固相萃取-电喷雾串联质谱（SPE-ESI-MS/MS）法对人参美白霜中人参皂苷进行定性鉴别。方法：通过SPE-ESI-MS/MS联用技术鉴定了人参美白霜中存在人参皂苷。结果：鉴定了人参美白霜中存在8种人参皂苷分别为人参皂苷Rb2,Rc,Rd,Re,Rg1,Rg2,Rh1和三七皂苷R2。结论：本方法简便、可靠,为添加人参皂苷成分的化妆品的质量控制提供了有效的方法。     

人参美白霜中人参皂苷的SPE—ESI—MS／MS分析

目的：应用固相萃取一电喷雾串联质谱（SPE—ESI—MS／MS）法对人参美白霜中人参皂苷进行定性鉴别。方法：通过SPE—ESI—MS／MS联用技术鉴定了人参美白霜中存在人参皂苷。结果：鉴定了人参美白霜中存在8种人参皂苷分别为人参皂苷Rb2,Rc,Rd,Re,Rg1,Rg2,Rh,和三七皂苷Rz。结论：本方法简便、可靠,为添加人参皂苷成分的化妆品的质量控制提供了有效的方法。     

HPLC/LTQ-FTMS用于白参中的丙二酰基人参皂苷的研究

在15mM 甲酸铵作为流动相添加剂条件下,采用HPLC/LTQ-FTMS 实现了白参样品中酸性人参皂苷的优化分离。根据高分辨质谱、多级质谱信息,结合保留规律验证,在白参萃取液中共检测和识别了八种丙二酰基人参皂苷,包括丙二酰基人参皂苷Rg1、Re、Rf、Rb1、Rc、Rb2、Rb3 和Rd。     

A Distinctive Pattern of Beauveria bassiana‐biotransformed Ginsenoside Products Triggers Mitochondria/FasL‐mediated Apoptosis in Colon Cancer Cells

Ginseng is one of the most commonly used adaptogens. Transformation into the minor ginsenosides produces compounds with more effective action. Beauveria bassiana, a teleomorph of Cordyceps bassiana, is a highly efficient producer of mammalian steroids and produces large amounts of sugar‐utilizing enzymes. However, the fermentation of steroid glycosides in ginseng with B. bassiana has never been studied. Thus, we evaluated the bioconversion of the major ginsenosides in white ginseng by B. bassiana. Interestingly, B. bassiana increased the total amount of protopanaxadiols and hydrolyzed Rb1 into minor ginsenosides, exhibiting high levels of Rd and Rg3, as well as moderate levels of Rb2 and Rc analyzed by high‐performance liquid chromatography coupled with evaporative light‐scattering detection. The β‐glucosidase activity was highly increased, which led to the selective elimination of sugar moiety at the 20‐C position of Rb1 to Rd, followed by Rg3. Rb2 and Rc accumulated because of the minimal activities of α‐L‐arabinopyranosidase and α‐L‐arabinofuranosidase, respectively. The fermentation product exerted dose‐dependent cytotoxicity in HCT‐15 cells, which are resistant to ginseng. The product, but not white ginseng, exhibited apoptotic effects via the Fas ligand and caspase 8/9. This study demonstrates for the first time that the B. bassiana‐fermented metabolites have potent apoptotic activity in colon cancer cells, linking to a therapeutic use. Copyright © 2015 John Wiley & Sons, Ltd.     

三七叶抗抑郁活性提取物PnGL中皂苷类成分分析

目的 建立三七叶抗抑郁活性提取物PnGL中皂苷类成分含量测定的方法.方法 采用香荚醛-高氯酸比色法,分别以人参二醇及人参皂苷Rb3为对照品,测定总皂苷的含量.采用HPLC法,Ultimate AQ-C18色谱柱,甲醇-水(69:31)为流动相,检测波长203 nm,柱温为25℃,同时测定PnGL中主要成分人参皂苷Rb1、Rb2、Rc、Rb3的含量.结果 以人参二醇及人参皂苷Rb3为对照品所测定的总皂苷含量,分别为97.25％、95.76％,差异不大.HPLC检测基线平稳,人参皂苷Rb1、Rb2、Rc、Rb3分离良好,分离度1.5以上,平均加样回收率分别为99.24 ％(RSD=1.63％)、96.41 ％(RSD=1.21％)、96.81％(RSD=0.86％)、100.16 ％(RSD=1.84％).结论 含量测定方法准确、重复性好,可为PnGL质量控制提供依据.     

西洋参中皂苷类成分提取方法对比

目的:选择更好的西洋参提取方法,提高人参皂苷的提取率。方法:以人参总皂苷、人参皂苷Rb1、Rb2、Rc、Rd、Re、Rg1、Rg2等7种主要单体皂苷收率为指标,用高效液相色谱法进行含量测定,比较热回流提取、超声波提取、罐组式逆流提取、组罐式超声波提取四种方法各自的优劣。结果:组罐式超声波提取是最佳的提取方法,它可以在保证提取效率的前提下,减少溶剂的用量,降低提取温度。结论:组罐式超声波提取法可望广泛地应用在制药工业上。     

Effects of ginsenosides,lectins and Momordica charantia insulin-like peptide on corticosterone production by isolated rat adrenal cells

Ginsenosides Rb1, Rb2, Rc and Rg1 inhibited steroidogenesis induced by a maximally active dose of corticotropin in isolated rat adrenal cells. The galactose-binding lectins from Momordica charantia seeds and Trichosanthes kirilowii tubers and mannose-binding concanavalin A did not affect basal corticosterone production. The lectins potentiated steroidogenesis induced by a submaximal dose of corticotropin but were without effect on steroidogenesis induced by a maximally active dose of corticotropin. Momordica charantia insulin-like peptide did not affect steroidogenesis.     

人参四逆汤抗休克作用的有效组分成分分析

目的 分析人参四逆汤抗失血性休克作用的提取组分S－1和S－7的组成成分。方法 结合硅胶色普柱分离，利用ESI／MS^n、MALDI－TOF／MS等技术分析鉴定S－1和S－7的化学成分。结果 从人参四逆汤水煎液的抗休克作用的有效组发S－7中分析和鉴定了人参皂苷－Ra1、Ra2、－Rb1、－Rb2、－Rb3、－Rc、－Rd、－Re、－Rg1、－Rg2、－Rg2、－Rf等12种人参皂苷；从有效组分S－1中检出下列二萜生物碱成分；苯甲酰乌头碱油酸酯（14－benzoylhypaconine－8－linoleate，HAL）、苯甲酰去氧乌头碱油酸酯（14－benzoyldeoxyaconine-8-oleate,HAO）、苯甲酰次乌头碱棕桐酸酯（14－benzoylhpaconine-8-palmitate，HAP）、苯甲酰中乌头酸（benzoylmesaconitine，BM）、苯甲酰乌头碱（benzoyla-conitine，BA）、苯甲酰次乌头碱（benzoylhpaconitine，BH）。结论 首次分析鉴定了人参四逆汤水煎液抗失血性休克作用的提取组分的化学成分。     

Triterpene saponins from Vietnamese ginseng (Panax vietnamensis) and their hepatocytoprotective activity

The methanol extract of Vietnamese ginseng (Panax vietnamensis) was found to possess hepatocytoprotective effects on D-galactosamine (D-GalN)/tumor necrosis factor-alpha (TNF-alpha)-induced cell death in primary cultured mouse hepatocytes. Further chemical investigation of the extract afforded two new dammarane-type triterpene saponins, ginsenoside Rh(5) (1) and vina-ginsenoside R(25) (2), as well as eight known dammarane-type triterpene saponins, majonoside R(2) (3), pseudo-ginsenoside RT(4) (4), vina-ginsenosides R(1) (5), R(2) (6), and R(10) (7), ginsenosides Rg(1) (8), Rh(1) (9), and Rh(4) (10), and a known sapogenin protopanaxatriol oxide II (11). Their structures were elucidated on the basis of spectral analysis. In addition, by the using LC-electrospray ionization (ESI)-MS method, five known saponins, ginsenosides Rb(1), Rb(2), Rc, Rd, and Re (12--16), were also identified in the extract. Among the compounds isolated, majonoside R(2) (3), the main saponin in Vietnamese ginseng, showed strong protective activity against D-GalN/TNF-alpha-induced cell death in primary cultured mouse hepatocytes. This demonstrates that the hepatocytoprotective effect of Vietnamese ginseng is due to dammarane-type triterpene saponins that have an ocotillol-type side chain, a characteristic constituent of Vietnamese ginseng.     

基于高分离度和对照图谱的红参中9种人参皂苷类成分“一测多评”质量评价研究

采用反相高效液相色谱法,Ultimate~C_(18)色谱柱,对红参中9种人参皂苷类成分进行了具有高分离度和高色谱峰纯度的同步色谱分离。采用"一测多评"中药质量评价模式,经一系列方法学考察,以人参皂苷Rb_1为内参物,建立了人参皂苷Rg_1,Re,Rf,Rb_1,Rc,Rb_2,Rb_3,Rd和20(S)-人参皂苷Rg3与人参皂苷Rb1间的相对校正因子与相对保留值,在红参对照图谱的辅助定位下,最终实现了仅采用人参皂苷Rb1一个中药对照品对红参中上述9种人参皂苷类成分的同步质量控制。     

Enrichment of the less polar ginsenoside (Rg3) from ginseng grown in New Zealand by post-harvest processing and extraction

Background: Previous studies showed that New Zealand-grown ginseng contains an abundance of ginsenosides and that rare less polar ginsenosides, such as Rg3, exhibit more pharmacological activities than polar ginsenosides, which are the major components of ginseng. Methods: The ginsenoside profile of New Zealand-grown Panax ginseng was manipulated by treatment with acetic acid, sodium hydroxide, pH, and high temperature. The abundance of 23 ginsenosides extracted by different treatments was quantified using high-performance liquid chromatography. Results:Treatment with 0.5 mol/L acetic acid can stimulate the degradation of polar ginsenosides to less polar ginsenosides (5.6％Rg3 was accumulated, P<0.0001). Furthermore, when ginseng root was treated at 121 ℃ for 100 min in a pH 3.0 acetic acid aqueous solution, the majority of the polar ginsenosides were converted into less polar ginsenosides. Specifically, 83.46 ± 3.69％(P= 0.0360) of the less polar ginsenosides and 41.01 ± 2.39％(P=0.0412) of Rg3 were enriched. In contrast, alkali treatment did not convert the polar ginsenosides into less polar ginsenosides at mild temperature and less conversion was observed compared with acid treatment at high temperature. Conclusion: This is the first attempt to manipulate the ginsenoside profile of New Zealand-grown ginseng. The conditions (high temperature with low pH) may be modified to produce and enrich the less polar ginsenoside fraction (especially Rg3) from the total ginseng extract.     

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??OBJECTIVE To compare the pharmacokinetic parameters of Xuesaitong dispersible tablets and common tablets in Beagle dogs. METHODS Using double cycle crossover trial, six Beagle dogs were treated with single oral dose of 100 mg of Xuesaitong dispersible tablets and conventional tablets and determining the pharmacokinetic parameters of ginsenoside Rb1 and Rg1 in Xuesaitong dispersible tablets and conventional tablets in Beagle dog plasma. RESULTS The ginsenoside Rb1 and Rg1 peak concentration of Xuesetong dispersible tablets in Beagle dog plasma was significantly higher than that of Xuesaitong tablets, the ginsenoside Rg1 peak time of Xuesaitong dispersible tablets in Beagle dog plasma was significantly earlier than that of Xuesaitong tablets. Additionally, the ginsenoside Rb1 peak time exhibited ahead of the trend, which is in line with the characteristics of rapid disintegration and absorption of preparation in vivo. CONCLUSION The plasma exposure in two preparation of ginsenoside Rb1 and Rg1 in Beagle dog holds fairly basic and no significant difference. But the ??_max of the main ingredients of Panax ginseng saponins Rb1 and Rg1 in Xuesaitong dispersed tablets, is significantly higher than that of the film coated tablets, and peak time is significantly shortened, which could promote the drug absorption.     

“一测多评”法测定腰痹通胶囊中5 种皂苷类成分的含量

目的：建立“一测多评”法测定腰痹通胶囊中皂苷类成分含量的分析方法。方法：以腰痹通胶囊为研究对象,建立参照物人参皂苷Rg1与三七皂苷R1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd的相对校正因子,分别采用外标法和“一测多评”法测定腰痹通胶囊中5种皂苷类成分的含量,并将两种测定方法的结果进行对比分析,以验证“一测多评”法的合理性、可行性和可重复性。结果：腰痹通胶囊中皂苷类成分间的相对校正因子分别为：f 人参皂苷Rg1/三七皂苷R1=0.999、f人参皂苷Rg1/人参皂苷Re=1.228、f人参皂苷Rg1/人参皂苷Rb1=0.990、f人参皂苷Rg1/人参皂苷Re=1.094。“一测多评”法的计算结果与外标法的实测值之间无显著性差异,实验所得的相对校正因子可信。结论：本实验建立的“一测多评”法可行准确,可以更合理、快速地实现腰痹通胶囊中多种皂苷类成分的质量控制。     

HPLC-光敏二极管阵列检测法测定人参单体皂甙的研究

讨论了流动相为异丙醇－甲醇非水体系配合极性柱分离人参单体皂甙的情况。试验中选用了极性较强的NH2基键合相桂，以甲醇－异丙醇（62：38V／V）为流动相，选择二极管阵列检测器，UV204um处检测，分离了单体皂甙Re、Rg1、Rb1，方法操作简单、快速、重现性好，线性范围宽，检出限低，可用于人参及各种制剂的分析。