Optimization of extraction process for total flavonoids of Sophorae Flos for the treatment of hyperlipidemia based on network pharmacology and molecular docking

This study aimed to investigate the mechanism of action of Sophora Flos(SF)in the treatment of hyperlipidemia(HLP)using network pharmacology and molecular docking methods,and to optimize the extraction process of the predicted active components.The STRING database was used for protein interaction analysis and PPI network construction via Cytoscape 3.9.1.Pymol was employed for docking and visualization.An extensive review of SF identifi ed 6 active ingredients,297 related objectives,84 disease objectives,and 57 total objectives.After protein interaction and topology analysis,18 core targets were identified.These included 146 gene function entries(P<0.05).Active compounds,mainly flavonoids,can modulate the expression of various proteins such as TNF,IL-6,IL-1β,PPARG,and TGFB1 to achieve therapeutic effects on HLP.The network pharmacology and molecular docking results suggested that the active fl avonoids component in SF may be related to the treatment of hyperlipidemia.Therefore,the orthogonal experiment method was used to optimize the extraction process of total fl avonoid from SF using ethanol refl ux extraction,based on a single factor experiment.The effects of refl ux time,solid-liquid ratio,ethanol concentration,and other factors on the extraction of total fl avonoid from SF were investigated.The optimum process conditions were refl ux time of 1.25 h,solid-liquid ratio of 1:15 g/mL and ethanol concentration of 60%.Using these conditions,the purity of total fl avonoid extracted from SF was 70.33±0.22%.

Microwave-assisted Extraction of Rutin and Quercetin from Flos Sophorae

Three microwave-assisted extraction(MAE) procedures were studied. The first procedure was household microwave oven dynamic extraction(HMODE). The second procedure was special microwave oven bath extraction(SMOBE). The third procedure was microwave resonant cavity dynamic extraction(MRCDE). The results obtained by the three microwave-assisted extraction procedures were compared with those obtained by using traditional Soxhlet extraction. The results indicate that the MAE not only took a shorter time, but also simplified the procedure, and made the extraction a higher yield. At the same time the results obtained by the three MAE procedures were also compared with each other.

山豆根的质量控制现状及质量标志物预测分析

山豆根含有生物碱、黄酮类、多糖、三萜及三萜皂苷、挥发油、有机酸等活性成分,主要有清热解毒、消肿利咽的功效。阐述山豆根的质量控制现状,预测山豆根的质量标志物。从传统药性、传统药效、不同复方配伍、可测性化学成分、新的药效用途、植物亲缘关系等7个方面对山豆根质量标志物作预测分析。山豆根的质量标志物可能含有苦参碱、氧化苦参碱、槐根碱、槐定碱、槐醇、芒柄花素、高丽槐素、金雀花碱、β-甾醇、红车轴草苷等化合物。这些化合物可首选作为山豆根的质量标志物,为建立山豆根的质量控制标准研究提供参考。

山豆根叶片光合特性对光强和干旱的响应

山豆根(Euchresta japonica)为我国Ⅱ级重点保护野生植物。该研究通过设置不同遮阴网的层数和采用不同浓度的聚乙二醇溶液浇灌,探讨了山豆根光合特性对光强和干旱的响应。结果表明,山豆根叶片的饱和光强为683.06~907.07μmol/(m~2·s);单层遮阴处理叶片的最大电子传递速率和最大净光合速率整体高于双层遮阴处理,其中单层遮阴且未进行干旱处理的叶片最大电子传递速率和最大净光合速率最高,分别为55.36和6.73μmol/(m~2·s);相同遮阴条件下,叶片的最大净光合速率及其对应的饱和光强均随干旱程度增加整体呈下降趋势;单层遮阴条件下的蒸腾速率和水分利用效率均整体高于双层遮阴条件下的。不同处理下叶片光系统Ⅱ的实际光化学效率、光化学猝灭系数以及非光化学猝灭系数等整体上均无显著差异(P>0.05)。山豆根属半阳生植物,其叶片利用弱光能力较强,植株具有较强的耐干旱能力。因此,建议在开展野外回归、迁地保护、人工栽培等工作时,进行适当遮阴处理并保持充足的土壤含水量。

山豆根菌核病病原菌鉴定及室内药剂筛选

采用常规组织分离法分离山豆根(Sophora tonkinensis Radix)菌核病病原菌,利用柯赫式法则进行致病性测定,基于形态特征、菌丝融合群测定、细胞核荧光染色以及rDNA-ITS序列分析进行病原菌的鉴定,并利用菌丝生长速率法测定5种杀菌剂对病原菌的抑制活性。结果表明,山豆根菌核病的病原菌为立枯丝核菌(Rhizoctonia solani),属于融合群AG-1。98%恶霉灵可溶性粉剂(SP)、75%肟菌·戊唑醇水分散粒剂(WG)、250 g/L吡唑醚菌酯乳油(EC)、10亿/g哈茨木霉菌悬浮剂(SC)、50%多菌灵可湿性粉剂(WP)对病原菌菌丝的生长均有较好的抑制效果,抑制中浓度(EC_(50))分别为0.0611、0.5757、0.0748、0.2843、0.7405 mg/L。

Optimization of Extraction Process of Quercetin from Flos Sophorae Immaturus Using Orthogonal Design

[Objectives]This study aimed to optimize the ultrasonic-assisted extraction process of quercetin from Flos Sophorae Immaturus.[Methods]The natural product quercetin in Flos Sophorae Immaturus was extracted by ultrasonic-assisted method,and the content of quercetin in the extract was determined by ultraviolet spectrophotometry.The effect of ethanol concentration,solid/liquid ratio,temperature,time and ultrasonic power on the yield of quercetin in Flos Sophorae Immaturus was investigated by using single-factor experiments.Based on the results of single-factor experiments,an orthogonal experiment was designed to optimize the extraction process.The experimental data obtained were subjected to range analysis,analysis of variance and SSR test using SPSS 20.0 software to obtain the optimal extraction process.The results concluded were verified.[Results]The optimal ultrasonic-assisted extraction process for quercetin in Flos Sophorae Immaturus was as follows:temperature of 85℃,time of 30 min,solid/liquid ratio of 1∶20 g/mL,ethanol concentration of 50%.Under the optimal extraction conditions,the yield of quercetin in Flos Sophorae Immaturus was the highest.[Conclusions]This study provides a theoretical basis for the application of quercetin in Flos Sophorae Immaturus.

基于专利视角的山豆根产业链开发现状评价与建议

从专利视角分析山豆根产业链并作出战略[优势(Strengths)、劣势(Weaknesses)、机会(Opportunities)和威胁(Threats),SWOT]评述,为其进一步开发提供了数据支撑和理论依据。对全球专利数据库进行详细检索并进行综合统计分析,从数量、质量、技术组成和分布等多个维度全面总结了山豆根专利的现状。其次,采用SWOT模型对山豆根专利开发现状进行了评价,并提出了相应的发展建议。结果表明,山豆根作为一种传统中药材,在医药、农业和食品等领域有着广泛的应用和开发价值,但也面临着种质资源保护、技术创新和知识产权保护等方面的挑战。期望通过本文的分析能够对山豆根产业链相关从业人员提供一定的参考和借鉴。

基于数据挖掘技术分析含山豆根组方的配伍用药规律研究

目的:研究含山豆根方剂的配伍用药规律,为山豆根的新药研发及临床应用提供参考。方法:收集整理药智网中山豆根药材中成药处方和中药方剂及中国知网中近15年明确含有山豆根处方的方剂,排除功效主治不明的处方,利用Excel 2019和“中医传承辅助平台(V2.5)”软件创建数据库进行药物频次统计、药物性味归经统计、关联规则分析和系统聚类等。结果:共纳入含山豆根方剂217首,用于治疗病症的类型共58种,排前4的病症分别为咽喉肿痛、感冒、胁痛、银屑病;涉及药物418味;常与山豆根配伍使用的药物频次高于20次的药物共23味,排前5的中药为甘草、玄参、桔梗、连翘、黄芩,配伍中药种类有补虚药、清热药,药性主要为寒、温、平。药味主要以苦、甘、辛居多,主要归脾、肝、胃、肺经。药物组合使用频次最多的为山豆根、桔梗、连翘、黄芩、金银花、射干、牛蒡子、薄荷、玄参、麦冬、甘草。结论:通过综合分析含山豆根方剂的主治病症、方剂类型、常配伍中药的使用频次、功效归属、性味归经及组方规律,可为山豆根的临证科学组方提供借鉴与参考。

山豆根中毒性脑病头颅影像特征

目的探讨山豆根中毒性脑病患者头颅影像学特点,为临床诊断提供依据。方法检索万方医学数据库、维普数据库、中国知网、中华医学期刊全文数据库自建库至2023年2月山豆根中毒性脑病文献,提取头颅影像学特点,建立数据并进行分析。结果研究纳入山豆根中毒性脑病患者87例,学龄前和学龄儿童多见(54/87,62.07%),临床症状表现为头晕、行走不稳、意识障碍、偏身扭转、全身乏力、球麻痹等,头颅影像主要表现:双侧小脑和/或双侧基底节区对称性病变,随中毒剂量增加,累及部位增多(如脑干、脑叶)。意识障碍多与双侧基底节区受累有关。头颅MRI表现为:对称性T1低信号、T2高信号,DWI高信号或等信号,ADC低信号或高信号。结论山豆根中毒性脑病影像学以双侧小脑、双侧基底节区对称性病变为特征,可累及脑干、脑叶,累及部位越多临床症状越重。

山豆根提取物质量标准研究

建立山豆根提取物的质量标准。选取乙醇浓度、料液比、回流提取时间及提取次数为影响因素,采用正交实验法优化山豆根提取物提取工艺,并以HPLC法测定山豆根提取物中苦参碱和氧化苦参碱的含量。优化提取工艺为:乙醇浓度60%、料液比1∶20(g∶mL)、回流时间2 h、提取次数3次。HPLC含量测定结果为:苦参碱及氧化苦参碱在选定范围内线性关系良好,加样回收率RSD不高于2%。提取工艺稳定可行,且建立的HPLC含量测定方法简便、快捷。

山豆根对植物病原菌的抑菌活性研究

【目的】目前对山豆根的研究大多集中在生物碱、多糖类和黄酮类等活性物质的药理学方面,对山豆根在植物病原菌的抑菌活性及活性物质方面的研究尚少,具有一定的研究价值。【方法】以我国农业重要植物病原菌为供试菌,采用有机溶剂冷浸提取法提取山豆根的有效成分,运用带毒平板法和菌丝生长速率法测定山豆根甲醇提取物及其各层萃取物对7种植物病原菌的抑菌活性,并采用液-液萃取分离法对山豆根甲醇提取物活性成分进行初步分离。【结果】1)山豆根甲醇提取物(浓度为10 mg/mL)对甘蔗凤梨病菌、甘蓝黑斑病菌、根腐病菌和茶轮斑病菌的抑菌率均在80%以上,对玉米大斑病菌、贡柑链格孢杆菌和辣椒炭疽病菌的抑菌活性相对次之;2)山豆根甲醇提取物对甘蓝黑斑病菌的毒力最高,有效抑制中浓度(EC50)为0.027 9 mg/mL;3)山豆根的抑菌活性物质主要存在于正丁醇层和水层萃取物中。【结论】山豆根具有一定的抗菌性,而且可能具有多种抗菌作用方式,后续可采用色谱和光谱等技术对山豆根活性成分进行进一步分离、纯化和鉴定,再根据其活性成分的性质和结构加以改造或修饰,从而提高山豆根抑菌活性成分的抑菌率,也为进一步以山豆根为导向开发植物源抑菌剂奠定基础,为植物源新农药的开发利用添砖加瓦。

(±)山豆根酮A10和A1的合成及抗氧化活性

以2,4,6-三羟基苯乙酮一水合物和对羟基苯甲醛为起始原料,经C-异戊烯基化反应、羟基保护、羟醛缩合、环合,脱保护合成了具有抗炎效果的天然产物山豆根酮A10和EuchrenoneA1,总收率分别为15.9%和12.8%。实验表明,山豆根酮A10具有一定的抗氧化活性。

4例山豆根引起的不良反应病例分析

山豆根在我国有着悠久的使用历史,其药用价值较高,但同时也存在着一定的毒性效应。本文回顾性分析了2019年9月至2022年10月我院中药房接收的4例成人患者服用山豆根引起不良反应的病例资料,探究山豆根不良反应的特征、发生机制及预防措施,以期为临床合理、安全使用山豆根提供参考。

Study on possible synergistic anticancer cachexia effects of the main active compounds of Radix Sophorae flavescentis

Background:Cancer cachexia is a complex disease secondary to cancer,and no specific therapy for it has been found.The Chinese herb Kushen(Radix Sophorae flavescentis)is the dried root of Sophora flavescens Aiton,which has been widely applied in treating digestive and urinary inflammatory diseases.Matrine,one of the main components of Radix Sophorae flavescentis,can alleviate cancer cachexia.Methods:Compounds from Radix Sophorae flavescentis were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.The targets related to cancer cachexia were queried from the Therapeutic Target Database(http://db.idrblab.net),DisGeNET database(http://www.disgenet.org),and Search Tool for Interacting Chemicals database,related literature,and constructed cancer cachexia-protein network.Cancer cachexia-protein network was merged with compound-protein networks respectively using Cytoscape software as well as network topology data and key targets counting.Pathway enrichment analysis was conducted via the Database for Functional Annotation Bioinformatics Microarray Analysis.Protein crystal structures and compound structures were queried from RCSB and PubChem databases,respectively.Molecular docking was conducted using Discovery Studio software.Results:The anticancer cachexia compounds of Radix Sophorae flavescentis were screened as oxymatrine,matrine,and kurarinol,and targets such as BIRC2,TNF and STAT3 were found.The mechanisms of oxymatrine,matrine,and kurarinol have the characteristics of synergy and complementarity.Kurarinol has a mechanism similar to that of matrine,which includes the FoxO signaling pathway,insulin resistance and mTOR signaling pathway.TNF signaling pathway is a common signaling pathway of kurarinol,oxymatrine and matrine.Adipocytokine signaling pathway is the other common pathway of kurarinol and oxymatrine except for the TNF signaling pathway.Kurarinol can be successfully docked with CYCS,GPX2,BIRC7,etc.,and kushenol C can be successfully docked with IKBKB and PIK3CD.Conclusion:Kurarinol,matrine,oxymatrine,and kushenol C may be the key compounds of Radix Sophorae flavescentis treating cancer cachexia.Additionally,TNF signaling pathway is the key pathway for the synergistic action of kurarinol,matrine and oxymatrine.

管萼山豆根总黄酮的提取工艺优化及其抗氧化活性研究

以管萼山豆根的叶为原料,采用微波-超声波辅助低共熔溶剂(DES)提取总黄酮,筛选出较优的DES体系,在单因素试验的基础上,运用Plackett-Burman和响应面试验优化其提取工艺;并考察了最佳提取工艺下总黄酮对DPPH·、·ABTS^(+)和·OH的清除作用。研究结果表明:较优的DES溶剂体系为n(氯化胆碱)∶n(三乙二醇)=1∶3,总黄酮的最佳提取工艺条件为提取时间45 min、提取温度74℃、料液比1∶45(g∶mL)、微波功率300 W、DES含水量25%、超声波功率300 W,在该条件下叶中总黄酮得率为3.460%;分别采用乙酸乙酯、热乙酸乙酯(50±1)℃、大孔吸附树脂AB-8、NKA-9和D101分离总黄酮,其回收率分别为23.40%、28.09%、88.66%、71.55%和41.86%;AB-8大孔树脂分离所得总黄酮对DPPH·、·ABTS^(+)与·OH的半数抑制质量浓度(IC_(50))值分别为0.01705、0.01835和0.12330 g/L;总黄酮对DPPH·、·ABTS^(+)的清除率显著高于维生素C(V_(c)),而对·OH的清除率略低于V_(c)。进一步分析发现:优化条件下叶总黄酮得率(3.460%)高于茎(2.059%)和根(1.875%),高于单一微波提取(3.377%)和超声波提取(3.390%),低于加热回流提取(3.635%),但所需时间仅为加热回流提取的1/4。

Optimization of the Extraction Process of Rutin from Flos Sophorae by Orthogonal Experiments

[Objectives]This study was conducted to optimize the extraction process of rutin in Flos Sophorae.[Methods]With the percentage content of rutin extracted from Flos Sophorae as the evaluation index and rutin as the reference substance,the content of rutin in Flos Sophorae extracted ultrasonically was determined by high performance liquid chromatography.L_(9)(3^(4))orthogonal experiments were carried out to optimize the three factors(solvent concentration,material-to-liquid ratio,extraction time)that affect the effect of ultrasonic extraction of rutin from Flos Sophorae.[Results]The extraction effect of the crude medicinal powder was best with the methanol concentration of 80%,the ratio of material to liquid at 0.1∶30,and the extraction time of 40 min.[Conclusions]This study provides a reference for optimizing the extraction process of rutin in Flos Sophorae.

山豆根ISSR-PCR反应体系优化及引物筛选

以山豆根为试验材料,采用正交试验考察DNA、引物、Mg^(2+)、dNTPs、Taq酶5个因素对ISSR-PCR反应体系的影响,建立并优化山豆根ISSR-PCR反应体系条件。结果表明:山豆根ISSR-PCR反应体系(20μL)最优条件为Mg^(2+)浓度3 mmol/L,dNTPs 0.2 mmol/L,Taq酶1 U,引物0.5μmol/L,模板DNA 100 ng,运用该体系从100条UBC引物中筛选到10条清晰、多态性好的引物。采用正交试验设计可有效建立山豆根ISSR-PCR反应的最优体系,为山豆根的亲缘关系,种质资源遗传多样性评价及品种鉴定和系统分类等研究提供科学依据。

槐树(Sophora japonica L.)种质资源现状及研究进展

对槐树种质资源在我国的分布和现状,以及山东、山西、河北和河南等地的资源调查收集情况进行了综述;从表型、细胞和生理生化、分子水平3个方面,对槐树种质资源评价的研究进展进行了归纳;以选择育种、人工杂交授粉和现代生物技术育种为例,对种质资源创新利用情况进行了总结,并对工作中存在的问题和发展前景进行了分析。

基于指纹图谱研究的山豆根药材、饮片、标准汤剂、配方颗粒比较研究

目的研究山豆根药材、饮片、标准汤剂、配方颗粒的主要成分检测方法,为山豆根配方颗粒的应用提供参考。方法通过高效液相色谱建立山豆根药材、饮片、标准汤剂、配方颗粒指纹图谱检测方法,利用相似度评价、相关性分析、聚类分析及主成分分析进行评价和分析。结果山豆根药材、饮片、标准汤剂和配方颗粒的指纹图谱色谱中均有5个共有特征峰,且全部指认;相似度评价及化学模式识别研究表明四者无明显的差异。结论山豆根药材、饮片、标准汤剂和配方颗粒的主要组成成分基本一致,可为山豆根配方颗粒的基础研究及临床应用研究提供参考。

Comparison on the Origin and Development of Sophora flavescens in China and Japan

Sophora flavescens , which was first recorded in Shen Nong s Herbal Classic , has the functions of clearing away heat, drying dampness, killing parasites and promoting urination. In its long-term application, traditional Chinese medicine has gradually deepened its understanding of the origin and efficacy of S. flavescens . In order to explore changes in the origin and efficacy of traditional Chinese herb S. flavescens in the history of China and Japan, in this paper, the origin and development of S. flavescens in China and Japan were compared from three aspects: the origin, medicinal efficacy, and processing methods of S. flavescens in China and Japan, hoping to provide clear understanding of the medicinal plant S. flavescens .