Screening hepatoprotective effective components of Lonicerae japonica Flos based on the spectrum-effect relationship and its mechanism exploring

Lonicerae japonicae Flos(LF)is a kind of healthcare food with hepatoprotective function.This study was designed to explore the spectrum-effect relationships between UPLC fingerprints and the hepatoprotective effects of LF.Fingerprints of ten batches of LF were established by UPLC-PDA.The inhibitory levels of AST and ALT were used as pharmacological indexes,and secoxyloganin,isochlorogenic acid A and isochlorogenic acid C were screened as hepatoprotective active compounds by grey relational analysis(GRA)and partial least squares regression analysis(PLSR).Caspase-3 was obtained by network pharmacology as a key target of hepatoprotective active compounds.Molecular docking is used to explore the interaction between small molecules and proteins.This work provided a general model of the combination of UPLC-PDA and hepatoprotective effect to study the spectrum-effect relationship of LF,which can be used to considerable methods and insight for the fundamental research of the material basis of similar healthcare food.

Comparative analysis of volatile constituents between herbal pair flos lonicerae-caulis lonicerae and its single herbs

Gas chromatography-mass spectrometry(GC-MS) and the chemometric resolution method(alternative moving window factor analysis,AMWFA) were used for comparative analysis of volatile constituents in herbal pair(HP) flos lonicerae-caulis lonicerae(FL-CL) and its single herbs.The temperature-programmed retention index(PTRI) was also employed for the identification of compounds.In total,44,39,and 50 volatile chemical components in volatile oil of FL,CL and HP FL-CL were separately determined qualitatively and quantitatively,accounting for 87.22%,94.54% and 90.08% total contents of volatile oil of FL,CL and HP FL-CL,respectively.The results show that there are 32 common volatile constituents between HP FL-CL and single herb FL,33 common volatile constituents between HP FL-CL and single herb CL,and 10 new constituents in the volatile oil of HP FL-CL.

Application of near-infrared spectroscopy for the rapid analysis of Lonicerae Japonicae Flos solution extracted by water

A rapid quantitative analytical method for three components of Lonicerae Japornicae Flos solution(Lonicera Japonica Thumb.)extracted by water was developed using near-infrared(NIR)spectroscopy and the partial least-squares(PLS)method.The NIR spectra of 81 samples collected from a production line were obtained.The concentrations of secologanic acid,chlorogenicacid and galuteolin were detemmined by using high-performance liquid chromatography-diodearray detection as the reference method.Several pretreatment methods for the NIR spectra wereusedi during PLS calibration.The most appropriate latent variable number of the PLS factor wasselected based on the standard error of cross-validation(SECV).The performance of the finalPLS models was evaluated according to SECV,standard error of predliction(SEP)and deter-mination coeficient(R^(2)).The compounds secologanic acid,chlorogenic acid and galuteolin hadSEP values of 0.030,0.061 and 1.668μg/mL,respectively and R^(2) values over 0.85.This workshows that NIR spectroscopy is a rapid and convenient method for the analysis of LoniceraeJaponicae Flos solution extracted by water.The proposed method can help in the application ofprocs analytical technology in the pha maceutical industry,particularly in tra ditional Chinesemedicine injections.

Quality Analysis and High-Performance Liquid Chromatographic Fingerprint Analysis of New Cultivated Kind of <i>Lonicerae japonicae</i>Flos “Hua Jin 6” from Different Harvest Times

Lonicerae japonicaeFlos (LJF) is widely used in traditional Chinese medicines for the treatment of various diseases, which is now in great demand every year and has a broad development prospect. However, the flowering phase of common LJF varieties is so short, which seriously restricts the development of LJF industry. As a new cultivated kind of Lonicerae japonicae Flos, “Hua Jin 6” has characteristics in long flowering phase and conveniently picking, which makes it have a broad development prospect. The aim of this study is to provide scientific guidance for its suitable harvest period by measuring yield and quality of “Hua Jin 6” from different harvest time. Studies show that flower size had a slowly rising trend from the first day to the seventh day, and then slowly declined or kept stable. There were no significant differences of total phenolic acid contents in different samples from different days, but contents of total flavonoids were on the rise and up to maximum in the ninth day. The contents of total iridoids had an increasing tendency from the first day to the fifth day and then kept relatively stable in other days. We demonstrated that the quality of “Hua Jin 6” is relatively stable and suitable for harvesting in all flower buds white stage in term of HPLC fingerprints. Our findings can make it possible to select the suitable time for different harvest purpose.

In vivoand in vitroanti-arthritic efficacy of a herbal formula consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos

OBJECTIVE Rheumatoid arthritis(RA)is the most common inflammatory autoimmune disease,affecting around 1% of the world population.Toll-like receptor 4(TLR4)signalling has been found to be involved in the pathogenesis of RA.It is a potential therapeutic target for RA treatment.A herbal formula(RL)consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos has traditionally been used in treating various inflammatory disorders.In this study,we would evaluate the anti-arthritic effect of RL on collagen-induced arthritis(CIA)in rats and investigate the involvement of TLR4 signaling in the mode of action of RL in vivo and in vitro.METHODS In vivo anti-arthritic efficacy was evaluated using CIA rats induced by bovine typeⅡ collagen.The treatment groups were treated with various concentrations of RL or positive control indomethacin for 35 d.Clinical signs(hind paw volume and arthritis severity scores),changes in serum inflammatory mediators,histological and radiographic changes of joints were investigated.Spleens and peritoneal macrophages were used to determine the effects of RL on innate and adaptive immune responses in CIA rats.The involvement of TLR4 signalling pathways in the anti-arthritic effect of RL was examined in cartilage tissue of CIA rats,murine RAW264.7macrophages and human THP-1 monocytic cells.RESULTS The severity of arthritis in the CIA rats was significantly attenuated by RL.Histological score and radiographic score were efficiently improved by RL.RL could also dose-dependently inhibit pro-inflammatory cytokines in serum of CIA rats.RL significantly inhibited the production of various pro-inflammatory mediators,the expression and/or activity of the components of TLR4 signalling pathways in animal tissue and cell lines.CONCLUSION RL possesses anti-arthritic effect on collagen-induced arthritis in rats.The therapeutic effect of RL may be related to its inhibition on pro-inflammatory cytokines in serum.The inhibition of the TLR4/TAK1/NF-κB and TLR4/TAK1/MAPK pathways participate in the anti-arthritic effects of RL.This provides a pharmacological justification for the use of RL in the control of various arthritic diseases.Further investigation should be done to develop RL into a modern anti-arthritic agent.

Systematic review for geo-authentic Lonicerae Japonicae Flos

In traditional Chinese medicine, Lonicerae Japonicae Flos is commonly used as anti-inflammatory, antiviral, and antipyretic herbal medicine, and geo-authentic herbs are believed to present the highest quality among all samples from different regions. To discuss the current situation and trend of geo-authentic Lonicerae Japonicae Flos, we searched Chinese Biomedicine Literature Database, Chinese Journal Full-text Database, Chinese Scientific Journal Full-text Database, Cochrane Central Register of Controlled Trials, Wanfang, and PubMed. We investigated all studies up to November 2015 pertaining to quality assessment, discrimination, pharmacological effects, planting or processing, or ecological system of geo-authentic Lonicerae Japonicae Flos. Sixty-five studies mainly discussing about chemical fingerprint, component analysis, planting and processing, discrimination between varieties, ecological system, pharmacological effects, and safety were systematically reviewed. By analyzing these studies, we found that the key points of geo-authentic Lonicerae Japonicae Flos research were quality and application. Further studies should focus on improving the quality by selecting the more superior of all varieties and evaluating clinical effectiveness.

Comprehensive analysis of the chemical constituents in sulfur-fumigated Lonicerae Japonicae Flos using UHPLC-LTQ-Orbitrap mass spectrometry

Lonicerae Japonicae Flos(LJF),a kind of traditional Chinese medicines(TCMs),has functions of detoxifying and evacuating heat.In the study,a method based on ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS)was developed for the chemical constituent analysis of organic acids,flavonoids,iridoids and new-generated compounds in sulfur-fumigated LJF(SF-LJF).Based on the accurate mass measurement(<±5 ppm),chromatographic behavior and diagnostic product ions(DPIs),113 constituents were unambiguously or tentatively characterized from SFLJF extract,including 46 chlorogenic acids,19 flavonoids,29 iridoid glycosides and 19 newly-generated compounds(including 17 sulfur-containing derivatives).In addition,5-CQA(5-caffeoylquinic acid,chlorogenic acid)was chosen to be sulfur-fumigated for the result validation.It was found that the most significant change of LJF after sulfur fumigation was the occurrence of sulfate or sulfite esterification reactions,which resulted in the emergence of many new sulfur-containing components.Our results demonstrated that the established method was a useful and efficient analytical tool to comprehensively characterize the material basis of SF-LJF,and also an excellent guidance of quality control about LJF.

Comparison of anti-inflammatory effects of Lonicerae Japonicae Flos and Lonicerae Flos based on network pharmacology

Objective:In Chinese herbal medicine(CHM)history,Lonicerae Japonicae Flos and Lonicerae Flos were used clinically as one drug,but now they are admitted as two herbal medicines in Chinese Pharmacopoeia(2010 edition).This study used network pharmacology to investigate whether the two can be used interchangeably for the treatment of inflammatory diseases in TCM clinical practice.Methods:Lonicerae Japonicae Flos and Lonicerae Flos were compared in the inflammation mechanism including core targets,Gene Ontology(GO),pathway and principle chemical components by the method of network pharmacology.Results:Lonicerae Japonicae Flos and Lonicerae Flos shared in six targets accounting for 66.7%of the entire core targets and more than half of the GO terms and pathways are similar.Organic acids are dominent compounds responsible for anti-inflammatory effects.Three of the compounds that bind to core targets including luteolin,quercetin and kaempferol,are shared in both herbs.Conclusion:Due to high similarity between Lonicerae Japonicae Flos and Lonicerae Flos,we believe that they can be used interchangeably for the inflammation in clinical treatment.

Plant metabolomics for studying the effect of two insecticides on comprehensive constituents of Lonicerae Japonicae Flos

Pesticides'overuse and misuse have been reported to induce ingredient variations in herbal medicine,which is now gaining attention in the medicinal field as a form of alternative medicine.To date,available studies on pesticide-induced ingredient variations of herbal medicine are limited only on a few compounds and remain most others unexamined.In this study,a plant metabolomics-based strategy was performed to systematically explore the effects of two frequently used insecticides on the comprehensive constituents of Lonicerae Japonicae Flos(LJF),the flower buds of Lonicera japonica Thunb..Field trials were designed on a cultivating plot of L.japonica with controls and treatments of imidacloprid(IMI)and compound flonicamid and acetamiprid(CFA).Unbiased metabolite profiling was conducted by ultra-high performance liquid chromatography/quadrupole-Orbitrap mass spectrometer.After data pretreatment by automatic extraction and screening,a data matrix of metabolite features was submitted for statistical analyses.Consequently,29 metabolic markers,including chlorogenic acids,iridoids and organic acid-glucosides were obtained and characterized.The relative quantitative assay was subsequently performed to monitor their variations across flowering developments.This is the first study that systematically explored the insecticide-induced metabolite variations of LJF while taking into account the inherent variability of flowering development.The results were beneficial for holistic quality assessment of LJF and significant for guiding scientific use of pesticides in the large-scale cultivation.

傅里叶变换红外光谱技术对金银花中有效成分定量模型建立及含量测定

中药材“安全、稳定、有效、可靠”是中医药现代化发展的现实需求,针对中药材质量良莠不齐的实际情况探索快速、精准、无损新型含量测定方法是目前中药材质量控制领域亟待解决问题之一。金银花有效成分复杂、质量波动较大,利用傅里叶变换红外光谱技术对金银花有效成分进行高效、无损快速测定可能是推动其质量控制的有效措施。该研究将HPLC法与傅里叶变换红外光谱技术相结合,运用化学计量学方法,建立金银花有效成分的含量测定模型,以期为金银花的快速、精准含量测定提供新方法。利用傅里叶变换光谱仪采集了河南、河北、山东三个产地共64份金银花的红外光谱,使用高效液相色谱(HPLC)法同时测定金银花中绿原酸、当药苷、断氧化马钱子苷、木犀草苷、异绿原酸A、异绿原酸C六种有效成分的含量。使用TQ分析软件中多元散射校正(MSC)、标准正态变换(SNV)、偏最小二乘法(PLS)、逐步多元线性回归(SMLR)、一阶导数(1st)、二阶导数(2nd)及SG卷曲平滑和诺里斯导数滤波(ND)进行光谱处理建立模型。经过对河南、河北、山东共64份金银花样品的化学计量学分析,结果表明PLS+MSC+2nd Der+NS对绿原酸含量测定效果最好,相关系数为0.7549,平均绝对偏差为0.24%;PLS+MSC+1st Der+ND对当药苷的含量测定效果最好,相关系数为0.9369,平均绝对偏差为0.00%;PLS+MSC+1st Der+SG对断氧化马钱子苷含量测定效果最好,相关系数为0.9678,平均绝对偏差为-0.06%;PLS+MSC+1st Der+NS、PLS+MSC+1st Der+SG以及PLS+SNV+1st Der+SG对木犀草苷的含量测定效果最好,相关系数为0.8590,平均绝对偏差为0.01%;PLS+MSC+2nd Der+ND对异绿原酸A的含量测定效果最好,相关系数为0.9339,平均绝对偏差为0.11%;PLS+MSC+2nd Der+SG对异绿原酸C的含量测定效果最好,相关系数为0.8661,平均绝对偏差为0.01%。通过傅里叶变换光谱技术建立对金银花有效成分的定量模型可以实现对未知金银花有效成分的含量测定,该研究为金银花有效成分的快速、无损测定提供了新的方法依据,有助于推动中药材质量的稳定、可控。

基于液质联用技术比较郁香忍冬花蕾与金银花化学成分

目的 比较郁香忍冬花蕾与金银花的成分差异,评估郁香忍冬花蕾药用价值,为其开发利用提供参考。方法 以70%甲醇为提取溶剂,以UPLC-TOF-MS分析成分种类,以HPLC-QQQ-MS测定成分含量,经多元统计分析确定成分差异。结果 在郁香忍冬花蕾与金银花中共鉴定出52种成分,20种定量成分含量差异显著,金银花有4种差异成分,郁香忍冬花蕾的异绿原酸C、阿魏酸、木犀草苷、芦丁含量是金银花的2倍以上,马钱酸、马钱苷含量分别是金银花的11.96、37.23倍。马钱苷、异绿原酸A、马钱酸、芦丁及断马钱酸是区分郁香忍冬花蕾及金银花的关键差异成分。结论 郁香忍冬花蕾与金银花成分种类相近,但含量差异明显,郁香忍冬花蕾具有重要药用价值,可进一步开发利用。

银翘的本草考证

银翘是金银花和连翘的合称。两者都是清热解毒药物,相须为用起到协同作用,提高疗效。作者通过对历代古籍及权威药学专著的收集、整理、比较、分析,分别对连翘和金银花的名称、性味归经、功效主治以及用药部位进行考证,结果发现历代本草著作对连翘、金银花记载不尽相同。连翘早期名为异翘,性味由“苦、平”转为“味苦性寒或微寒”,用药部位由根部发展为全叶,具有消肿排脓、清热解毒、疏散风热的功效。忍冬为金银花早期用名,只采用茎叶作为用药部位到茎、叶、花均可入药,属“寒”性药物,具有清热解毒之功效。考证结果可为今后成方制剂开发及临床应用提供重要参考。

GC-MS指纹图谱技术结合多元统计分析对“银花”类药材的鉴定

本文通过探究金银花、山银花和川银花3种“银花”类药材次生代谢物种类与含量的差异,为鉴定“银花”类药材提供科学依据。采用GC-MS技术结合主成分分析、正交偏最小二乘判别分析(OPLS-DA)等多元统计分析方法进行模式识别。成功构建OPLS-DA模型并筛选得到19种差异代谢物,且3种“银花”类药材代谢物种类和含量存在较大差异。基于GC-MS指纹图谱技术结合多元统计分析的方法适用于3种“银花”类药材的鉴定,为该类药材的质量评价提供参考。

不同前处理方法对5种中药农药残留量检测的影响

目的:研究分析不同前处理方法对当归、菊花、枸杞子、黄芪、金银花中农药残留测定结果的影响。方法:样品分别经QuEChERS法和高速匀浆结合亲水亲油平衡(HLB)材料固相萃取法处理,采用超高效液相色谱-串联质谱法(UHPLC-MS/MS)同时测定克百威、3-羧基克百威、甲拌磷、甲拌磷砜、甲拌磷亚砜、甲基异柳磷等5种农药残留量。采用超高效液相色谱柱Agilent Poroshell 120 SB-C_(18)(2.1 mm×100 mm,2.7μm);采用梯度洗脱方式,水相为含5 mmol·L^(-1)甲酸铵的0.1%甲酸溶液,有机相为含5 mmol·L^(-1)甲酸铵的0.1%甲酸溶液-乙腈(体积比5∶95);柱温为35℃;质谱分析中以电喷雾离子源正离子模式扫描,多反应监测模式检测。结果:QuEChERS法和HLB固相萃取法测得结果偏差在9.09%~55.56%之间。结论:中药农药残留量检测前处理方法的选定,建议以阳性样本的测定数据为评价指标与依据,选择测定值较高、提取效率高的方法。

基于网络药理学和分子对接探究金银花抗肺纤维化的作用机制

目的:基于网络药理学和分子对接技术探讨金银花抗肺纤维化的作用机制。方法:基于前期金银花代谢谱鉴定的活性成分和相关文献报道的成分,采用口服生物利用度(OB)≥30%,类药性(DL)≥0.18为阈值,借助TCMSP和SwissTargetPrediction数据库检索成分靶点;以“pulmonary fibrosis”“lung paralysis”“lung flaccidity”为关键词,通过GeneCards、TTD等数据库检索与肺纤维化相关的疾病靶点,绘制药物—疾病共有靶点韦恩图。利用STRING构建蛋白互作PPI网络,拓扑分析筛选核心靶点。运用Metascape对核心靶点进行GO功能分析和KEGG通路富集分析;并应用Cytoscape 3.7.1构建“金银花—成分—靶点—通路”网络;最后通过AutoDock_Vina将活性成分与核心靶点进行分子对接,验证结合力。结果:筛选得到24个潜在活性成分,挖掘436个成分靶点和364个疾病靶点,其中交集靶点93个,核心靶点有TP53、AKT1、MAPK1、STAT3等共17个。GO和KEGG分析发现金银花可能通过作用于病毒感染、癌症中的信号通路等关键信号通路来调控炎症反应和防御应答等。“金银花—成分—靶点—通路”网络图筛选核心化合物4个:木犀草素、槲皮素、山柰酚和金圣草素。分子对接进一步验证核心成分和关键靶点的相互作用,表明活性成分与其靶点有较好的对接活性。结论:网络药理学联用分子对接技术揭示金银花可以通过多成分、多通路、多靶点作用于肺纤维化,为其进一步的深入研究与临床应用提供了理论依据与新思路。

基于Heracles Neo超快速气相电子鼻快速鉴别金银花与山银花的研究

目的 通过Heracles Neo超快速气相电子鼻技术,对金银花与山银花的气味物质进行分析研究。方法 采用Heracles Neo超快速气相电子鼻对不同批次金银花与山银花的气味进行分析,获得其电子鼻气相色谱信息,通过筛选色谱峰判别能力≥0.850,峰面积≥1000的色谱峰,分析金银花与山银花可能的主要差异化合物,并建立金银花与山银花基于电子鼻色谱信息的主成分分析(principal component analysis,PCA)、判别因子分析(discriminant factor analysis,DFA)、软独立建模分析(soft independent modeling of class analogy,SIMCA)、浓度量化(concentration quantification,CQ)统计分析模型。结果 从山银花与金银花中筛选出三甲胺、乙醇、正丁胺等14种主要气味差异化合物,其中巴豆醛、2-甲基丁醇、顺式-3-己烯-1-醇、顺式-3-己烯醇甲酸酯、苯甲醛5种主要差异化合物色谱峰判别能力大于0.900,且色谱峰面积相对较大,表明其可能为判别金银花和山银花的关键性挥发性化合物。建立的PCA、DFA、SIMCA统计分析模型识别能力分别为91%、100%、100%,说明PCA、DFA、SIMCA统计分析模型可对金银花与山银花进行快速有效鉴别;CQ分析模型线性为0.9779,对于金银花中掺假山银花线性良好,说明CQ分析模型可以对金银花中掺假山银花量进行快速鉴别。结论 Heracles Neo超快速气相电子鼻技术可实现对金银花与山银花快速鉴别,为金银花原药材粉末及其金银花制剂中掺假山银花快速鉴定提供了一种新的科学方法 。

基于R语言的专利中药复方防治呼吸道传染病的用药规律研究

【目的】挖掘专利中药复方防治呼吸道传染病的遣方用药规律,为呼吸道传染病的防治提供参考。【方法】在国家专利数据库中收集防治呼吸道传染病的中药复方,经数据筛选与规范化后,利用R语言进行数据挖掘。【结果】共纳入429首中药专利复方,涉及中药846味;高频中药有26味,使用频次居前10位的药物分别为甘草、金银花、黄芩、连翘、桔梗、百部、苦杏仁、黄芪、柴胡、薄荷。高频中药主要为清热解毒药和补气药;所涉中药药性以寒为主,药味多为苦、甘、辛,主归肺经;关联规则分析得到19条核心关联规则、多个药物组合;聚类分析得到3个药物聚类。【结论】专利中药复方防治呼吸道传染病时注重寒温相配、宣降相伍,着重宣散肺邪、清热解毒,同时不忘益气和中、滋阴润燥,使祛邪不伤正、扶正不敛邪。

银翘解毒片(胶囊)中灰毡毛忍冬皂苷乙的检查研究

目的 建立银翘解毒片(胶囊)中灰毡毛忍冬皂苷乙的检查方法,考察是否存在山银花掺伪金银花情况。方法 以灰毡毛忍冬皂苷乙为检测指标,采用薄层色谱法(TLC)和高效液相色谱-蒸发光散射法(HPLC-ELSD)进行定性分析,并运用超高效液相色谱-质谱法(UPLC-MS/MS)进行结果确认。结果 在薄层色谱中,有3批样品出现与灰毡毛忍冬皂苷乙对照品位置和颜色一致的斑点,在液相色谱中,有4批次样品出现与灰毡毛忍冬皂苷乙对照品保留时间一致的色谱峰,通过UPLC-MS/MS法验证,4批样品检出灰毡毛忍冬皂苷乙成分,出现山银花掺伪情况。结论 所建方法能准确甄别银翘解毒片(胶囊)中灰毡毛忍冬皂苷乙的情况,有助于控制该制剂中金银花药味的质量。

金银花质量标准中木犀草苷含量测定方法探讨及不确定度分析

目的:探讨金银花现行质量标准中木犀草苷的含量测定方法的测量不确定度,为完善金银花药材及饮片的质量标准提供参考。方法:采用《中国药典》2020年版中金银花项下含量测定方法,测定金银花饮片中木犀草苷的含量,并评估方法的测量不确定度及其对检验结果判定的影响。结果:实验发现,按照《中国药典》2020年版四部高效液相色谱法的色谱参数调整要求,调整流动相比例,同批次金银花样品的木犀草苷含量测定结果存在差异。金银花中木犀草苷含量测定结果为(0.048±0.015)%(置信概率为95%,k=2)。结论:通过评估方法的测量不确定度发现,色谱参数调整是本实验贡献最大的不确定度分量,调整前后测定结果差异较大,并影响检验结果的符合性判定,金银花药材及饮片中木犀草苷的含量测定方法亟需完善和修订。

金银花质量控制研究进展

金银花是大宗常用药材,常用于治疗风热感冒、温病发热等症,其质量决定了临床用药的安全性和有效性,是产业发展的关键所在。查阅近5年金银花质量控制领域的研究成果,归纳金银花性状、显微、薄层及DNA分子标记的鉴别依据,并梳理多种高效液相色谱、超高效液相色谱、质谱、光谱等多种检测平台及方法的创新及其对提升金银花质量的推动作用,全面总结了金银花质量控制体系。可为金银花质量控制与评价提供参考和依据,指导农业生产促进质量提升,助力金银花产业高质量发展。