Immune Homeostasis: Effects of Chinese Herbal Formulae and Herb-Derived Compounds on Allergic Asthma in Different Experimental Models

Allergic asthma is thought to arise from an imbalance of immune regulation, which is characterized by the production of large quantities of Ig E antibodies by B cells and a decrease of the interferon-γ/interleukin-4(Th1/Th2) ratio. Certain immunomodulatory components and Chinese herbal formulae have been used in traditional herbal medicine for thousands of years. However, there are few studies performing evidence-based Chinese medicine(CM) research on the mechanisms and efficacy of these drugs in allergic asthma. This review aims to explore the roles of Chinese herbal formulae and herb-derived compounds in experimental research models of allergic asthma. We screened published modern CM research results on the experimental effects of Chinese herbal formulae and herb-derived bioactive compounds for allergic asthma and their possible underlying mechanisms in English language articles from the Pub Med and the Google Scholar databases with the keywords allergic asthma, experimental model and Chinese herbal medicine. We found 22 Chinese herb species and 31 herb-derived anti-asthmatic compounds as well as 12 Chinese herbal formulae which showed a reduction of airway hyperresponsiveness, allergen-specific immunoglobulin E, inflammatory cell infiltration and a regulation of Th1 and Th2 cytokines in vivo, in vitro and ex vivo, respectively. Chinese herbal formulae and herbderived bioactive compounds exhibit immunomodulatory, anti-inflammatory and anti-asthma activities in different experimental models and their various mechanisms of action are being investigated in modern CM research with genomics, proteomics and metabolomics technologies, which will lead to a new era in the development of new drug discovery for allergic asthma in CM.     

“形、色、气、味”鉴中药

"形、色、气、味"是每种中药材各种构成和成分性质的综合体现,运用"形、色、气、味"的技巧,"形状三看法"、"色泽三观法"、"气味三嗅法"、"味道三感法"鉴别的技能,既简单快捷,又准确无误,对指导中药鉴别工作,尤其是在没有理化实验条件的情况下,对中药鉴别有普遍的现实指导意义。     

UHPLC-Q-TOF/MS对藜芦药材化学成分的快速分离与鉴别

目的采用超高效液相色谱仪与四极杆飞行时间串联质谱仪联用技术(UHPLC-Q-TOF/MS)对藜芦中化学成分进行分析鉴别。方法应用Agilent Eclipse Plus C18色谱柱(2.1 mm×100 mm,1.8 μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相梯度洗脱,电喷雾离子源,正离子模式下采集数据。结果共鉴别出黎芦药材中32个化学成分,通过比较碎片离子对其中的3对同分异构体进行了结构解析。结论采用UHPLC-Q-TOF/MS方法能快速地在一张图谱上表征出藜芦中的主要化学成分,为藜芦的药效物质基础及化学成分体内代谢与作用机制的深入研究奠定了基础。     

Metabolomics-based comparative analysis of the effects of host and environment on Viscum coloratum metabolites and antioxidative activities

Viscum coloratum (Kom.) Nakai is a well-known medicinal hemiparasite widely distributed in Asia. The synthesis and accumulation of its metabolites are affected by both environmental factors and the host plants, while the latter of which is usually overlooked. The purpose of this study was to comprehensively evaluate the effects of host and habitat on the metabolites in V. coloratum through multiple chemical and biological approaches. The metabolite profile of V. coloratum harvested from three different host plants in two habitats were determined by multiple chemical methods including high-performance liquid chromatography-ultraviolet (HPLC-UV), gas chromatography-flame ionization detector (GC-FID) and ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF/MS). The differences in antioxidant efficacy of V. coloratum were determined based on multiple in vitro models. The multivariate statistical analysis and data fusion strategy were applied to analyze the differences in metabolite profile and antioxidant activity of V. coloratum. Results indicated that the metabolite profile obtained by various chemical approaches was simultaneously affected by host and environment factors, and the environment plays a key role. Meanwhile, three main differential metabolites between two environment groups were identified. The results of antioxidant assay indicated that the environment has greater effects on the biological activity of V. coloratum than the host. Therefore, we conclude that the integration of various chemical and biological approaches combined with multivariate statistical and data fusion analysis, which can determine the influences of host plant and habitat on the metabolites, is a powerful strategy to control the quality of semi-parasitic herbal medicine.     

Rapid and sensitive analysis of trace β-blockers by magnetic solid-phase extraction coupled with Fourier transform ion cyclotron resonance mass spectrometry

A rapid and sensitive method for analyzing trace β-blockers in complex biological samples, which involved magnetic solid-phase extraction (MSPE) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS), was developed. Novel nanosilver-functionalized magnetic nanoparticles with an interlayer of poly(3,4-dihydroxyphenylalanine) (polyDOPA@Ag-MNPs) were synthesized and used as MSPE adsorbents to extract trace β-blockers from biological samples. After extraction, the analytes loaded on the polyDOPA@Ag-MNPs were desorbed using an organic solvent and analyzed by FTICR-MS. The method was rapid and sensitive, with a total detection procedure of less than 10 min as well as limits of detection and quantification in the ranges of 3.5–6.8 pg/mL and 11.7–22.8 pg/mL, respectively. The accuracy of the method was also desirable, with recoveries ranging from 80.9% to 91.0% following the detection of analytes in human blood samples. All the experimental results demonstrated that the developed MSPE-FTICR-MS method was suitable for the rapid and sensitive analysis of trace β-blockers in complex biological samples.     

超临界CO2流体萃取与分子蒸馏联用技术提取分离川芎挥发性成分及其GC/MS分析

目的研究超临界CO2流体萃取(SFE)与分子蒸馏(MD)联用技术对中药挥发性成分提取与分离、纯化的可行性.方法采用超临界CO2流体萃取技术提取川芎的挥发性成分,再将所得到的挥发性成分进行分子蒸馏.以气相色谱(GC)-质谱(MS)技术分别测定萃取物和蒸馏物的化学成分,并对成分种类及其相对含量的变化进行比较.结果川芎的超临界CO2流体萃取物共鉴定了45种成分,而超临界CO2流体萃取物再经分子蒸馏后所得蒸馏物的成分种类有所减少,为39种,两提取物所含成分的含量也有所变化.结论SFE与MD联用技术可用于挥发性成分的分离、纯化,且优于单一SFE技术.     

超临界CO2流体萃取与分子蒸馏联用技术提取分离川芎挥发性成分及其GC／MS分析

目的 研究超临界CO2流体萃取（SFE）与分子蒸馏（MD）联用技术对中药挥发性成分提取与分离，纯化的可行性。方法 采用超临界CO2流体萃取技术提取川芎的挥发性成分，再将所得到的挥发性成分进行分子蒸馏，以气相色谱（GC）-质谱（MS）技术分别测定萃取物和蒸馏物的化学成分，并对成分种类及其相对含量的变化进行比较。结果 川芎的超临界CO2流体萃取物共鉴定了45种成分，而超临界CO2流体萃取物再经分子蒸馏后所得蒸馏物的成分种类有所减少，为39种，两提取物所含成分的含量也有所变化。结论 SFE与MD联用技术可用于挥发性成分的分离，纯化，且优于单一SFE技术。     

HPLC-TOFMS法对白花蛇舌草药材及其注射剂中化学成分的快速分离与鉴别

目的采用HPLC-TOFMS对白花蛇舌草药材及其注射剂中化学成分进行快速分离鉴别。方法色谱柱Ag-ilent Zorbax SB-C18柱(4.6mm×250mm,5μm),以0.3%醋酸(A)-甲醇(B)为流动相,梯度洗脱,0～30min,体积分数30%～90%B,进样量10μl,流速1.0ml/min,柱温为30℃,飞行时间质谱配有ESI离子源,质量数扫描范围m/z100～1000。结果采用HPLC-TOFMS方法对白花蛇舌草药材及其注射液制剂进行了在线化学成分分离与质谱表征,一次性在一张图谱上共鉴别出白花蛇舌草中11个化学成分,市售注射剂中6个成分,自制注射剂中2个成分。结论这可为中药白花蛇舌草及其注射剂的药效物质基础和质量控制研究奠定基础。     

江苏地产太子参代谢物的动态积累分析

目的 分析不同采收时间江苏地产太子参代谢产物的动态积累变化。方法 采用UPLC-Triple TOF-MS/MS技术对样品进行测定,结合Mass数据等对总离子流图主要分子离子峰进行归属,用SIMCA-P软件进行主成分分析。HPLC法测定太子参中指标性成分（主要代谢差异标志物）环肽A、B的含量。结果 共鉴定出12种化合物。主成分得分图显示,不同采收时间太子参样本能较明显地分开;通过载荷图筛选出差异显著的10种标志物,且这10种标志物呈现出不同的变化规律。太子参环肽A（HA）和环肽B（HB）分别在7月下旬和8月上旬含量较高。结论 为揭示不同采收时间太子参中代谢物积累动态规律及确定其药材合理采收期提供基础资料。      

An integrated strategy for comprehensive characterization of metabolites and metabolic profiles of bufadienolides from Venenum Bufonis in rats

Comprehensive characterization of metabolites and metabolic profiles in plasma has considerable significance in determining the efficacy and safety of traditional Chinese medicine (TCM) in vivo. However, this process is usually hindered by the insufficient characteristic fragments of metabolites, ubiquitous matrix interference, and complicated screening and identification procedures for metabolites. In this study, an effective strategy was established to systematically characterize the metabolites, deduce the metabolic pathways, and describe the metabolic profiles of bufadienolides isolated from Venenum Bufonis in vivo. The strategy was divided into five steps. First, the blank and test plasma samples were injected into an ultra-high performance liquid chromatography/linear trap quadrupole-orbitrap-mass spectrometry (MS) system in the full scan mode continuously five times to screen for valid matrix compounds and metabolites. Second, an extension-mass defect filter model was established to obtain the targeted precursor ions of the list of bufadienolide metabolites, which reduced approximately 39% of the interfering ions. Third, an acquisition model was developed and used to trigger more tandem MS (MS/MS) fragments of precursor ions based on the targeted ion list. The acquisition mode enhanced the acquisition capability by approximately four times than that of the regular data-dependent acquisition mode. Fourth, the acquired data were imported into Compound Discoverer software for identification of metabolites with metabolic network prediction. The main in vivo metabolic pathways of bufadienolides were elucidated. A total of 147 metabolites were characterized, and the main biotransformation reactions of bufadienolides were hydroxylation, dihydroxylation, and isomerization. Finally, the main prototype bufadienolides in plasma at different time points were determined using LC-MS/MS, and the metabolic profiles were clearly identified. This strategy could be widely used to elucidate the metabolic profiles of TCM preparations or Chinese patent medicines in vivo and provide critical data for rational drug use.     

Offline two-dimensional liquid chromatography coupled with ion mobility-quadrupole time-of-flight mass spectrometry enabling four-dimensional separation and characterization of the multicomponents from white ginseng and red ginseng

Inherent complexity of plant metabolites necessitates the use of multi-dimensional information to accomplish comprehensive profiling and confirmative identification. A dimension-enhanced strategy, by offline two-dimensional liquid chromatography/ion mobility-quadrupole time-of-flight mass spectrometry (2D-LC/IM-QTOF-MS) enabling four-dimensional separations (2D-LC, IM, and MS), is proposed. In combination with in-house database-driven automated peak annotation, this strategy was utilized to characterize ginsenosides simultaneously from white ginseng (WG) and red ginseng (RG). An offline 2D-LC system configuring an Xbridge Amide column and an HSS T3 column showed orthogonality 0.76 in the resolution of ginsenosides. Ginsenoside analysis was performed by data-independent high-definition MS^E (HDMS^E) in the negative ESI mode on a Vion™ IMS-QTOF hybrid high-resolution mass spectrometer, which could better resolve ginsenosides than MS^E and directly give the CCS information. An in-house ginsenoside database recording 504 known ginsenosides and 58 reference compounds, was established to assist the identification of ginsenosides. Streamlined workflows, by applying UNIFI™ to automatedly annotate the HDMS^E data, were proposed. We could separate and characterize 323 ginsenosides (including 286 from WG and 306 from RG), and 125 thereof may have not been isolated from the Panax genus. The established 2D-LC/IM-QTOF-HDMS^E approach could also act as a magnifier to probe differentiated components between WG and RG. Compared with conventional approaches, this dimension-enhanced strategy could better resolve coeluting herbal components and more efficiently, more reliably identify the multicomponents, which, we believe, offers more possibilities for the systematic exposure and confirmative identification of plant metabolites.     

中草药化学成分的研究

"中草药"是"中药"和"草药"的合成词,虽然"中药"和"草药"就其本质来讲皆源于天然的植物、动物和矿物以及人工创造的生物材料和仿生材料,但两者药用含义有很大的区别。"中药"是以中医药理论为指导使用的药物,有坚实的临床基础和确切的疗效,亦有源于临床实践的独特理论体系。尽管西方药学发展迅速并逐渐取代了世界上大多数国家的民族药学,但中药依然显示出强大的生命力,并正以历史上未曾有过的速度发展,因此,应坚信中草药中存在疾病预防和康复的物质基础。为了揭示中草药的物质基础,即中草药化学成分,人们尝试了许多方法和途径,各有长短,系统性分析和总结这些方法和途径对于中草药化学成分的深入研究和中草药资源的可持续利用皆有重要意义。     

应用快速色谱技术构建少数民族大复方组分库的方法学研究

目的 建立少数民族药大复方全组分的快速制备方法,为有效成分的高通量筛选提供大量样品。方法 应用快速色谱仪,以蒙药白脉散乙酸乙酯提取部位为研究对象,采用线性梯度洗脱法快速划段制备一系列色谱组分,采用高效液相色谱法对分离效果进行评价。结果　从该复方乙酸乙酯部位获得29个色谱组分,该方法对组方中不同性质的物质起到了较好的分离与富集效果。结论 快速色谱技术可用于构建民族复方药物组分库,这将有利于攻克民族药大复方药效物质基础研究难题。     

Impaired tricarboxylic acid cycle flux and mitochondrial aerobic respiration during isoproterenol induced myocardial ischemia is rescued by bilobalide

There is an urgent need to elucidate the pathogenesis of myocardial ischemia (MI) and potential drug treatments. Here, the anti-MI mechanism and material basis of Ginkgo biloba L. extract (GBE) were studied from the perspective of energy metabolism flux regulation. Metabolic flux analysis (MFA) was performed to investigate energy metabolism flux disorder and the regulatory nodes of GBE components in isoproterenol (ISO)-induced ischemia-like cardiomyocytes. It showed that [U–¹³C] glucose derived m+2 isotopologues from the upstream tricarboxylic acid (TCA) cycle metabolites were markedly accumulated in ISO-injured cardiomyocytes, but the opposite was seen for the downstream metabolites, while their total cellular concentrations were increased. This indicates a blockage of carbon flow from glycolysis and enhanced anaplerosis from other carbon sources. A Seahorse test was used to screen for GBE components with regulatory effects on mitochondrial aerobic respiratory dysfunction. It showed that bilobalide protected against impaired mitochondrial aerobic respiration. MFA also showed that bilobalide significantly modulated the TCA cycle flux, reduced abnormal metabolite accumulation, and balanced the demand of different carbon sources. Western blotting and PCR analysis showed that bilobalide decreased the enhanced expression of key metabolic enzymes in injured cells. Bilobalide''s efficacy was verified by in vivo experiments in rats. This is the first report to show that bilobalide, the active ingredient of GBE, protects against MI by rescuing impaired TCA cycle flux. This provides a new mechanism and potential drug treatment for MI. It also shows the potential of MFA/Seahorse combination as a powerful strategy for pharmacological research on herbal medicine.     

关于草药标准化的研究综述

由于自然产品有化学多样性的特点,使之具有很高的实用性,无论是作为纯天然化合物或是标准化植物提取物,它们都为新药的开发提供了很多可能。与现代西药比较,草药通常被用于治疗慢性疾病。规范化操作可保证植物的一个或多个活性成分及化合物的含量。在新药研发方面,植物药的开发成本较低且资源丰富,但植物的生长环境和基因等因素会在很大程度上影响植物提取物的生物化学成分。植物药的生产要求源植物在基因方面的单种栽培完全符合标准条件,以保证其生物化学指标的一致性,并可以优化植物药的安全性和有效性。本文旨在讨论如何改进植物药提取的规范化操作方法以及如何有效制备质量及效果稳定的植物药。     

检测中草药提取物中黄曲霉毒素的高灵敏方法

目的建立中草药提取物中黄曲霉毒素B1,B2,G1,G2的高灵敏测定方法。方法样品经提取和过滤后用免疫亲和柱净化和衍生,然后采用不同比例(15%~30%)乙腈/水梯度洗脱,用具荧光检测器的高效液相色谱仪检测。结果检出限:黄曲霉素G1为0.09 ppb;B2为0.02 ppb;B1为0.03 ppb;G2为0.04 ppb,均低于文献报道的检出限,样品加标回收率为90%~103.2%。结论该方法简单、快速、准确、灵敏度高,可用于中药中黄曲霉毒素含量的测定。     

HS/SPME-GC-MS法分析不同采收期薄荷药材挥发油成分动态变化

目的基于顶空固相微萃取-气相色谱-质谱联用(HS/SPME-GC-MS)分析法对不同采收期薄荷药材样品中挥发油成分进行研究。方法采用顶空固相微萃取法提取挥发油成分,GC-MS进行分析鉴定,应用峰面积归一化法定量。结果成熟期薄荷药材共检出47种化合物,定性并定量12种化合物,总含量为78.23%。对薄荷生物合成途径中的关键成分左旋柠檬烯、薄荷酮和左旋薄荷醇进行不同采收期的对比,初步确定该种薄荷的最佳采收期为9月。结论该方法可以高效、快速地分析薄荷药材的挥发油成分,为揭示不同采收期薄荷药材中挥发油成分动态变化及确定薄荷药材合理采收期提供基础资料。     

涉及中草药中毒的法医学检验鉴定程序及方法

目前医疗纠纷或其他案(事)件中涉及中草药中毒鉴定的数量日益增多，而法医学检验中的中草药的检验鉴定难度较大．介绍了经反复实践验证可行的中草药五步法医学检验鉴定程序和方法，对案件所涉及的中草药的毒性和毒理作用作出客观综合的认定与评价．