CN113156011A - 一种小柴胡颗粒活性成分群的构建方法 - Google Patents
一种小柴胡颗粒活性成分群的构建方法 Download PDFInfo
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Abstract
本发明涉及一种小柴胡颗粒活性成分群的构建方法。该构建方法包括以下步骤:取小柴胡颗粒浸膏样品,以甲醇水溶液为溶剂,制备供试品溶液;取所述供试品溶液进行液相色谱质谱法分析,得到小柴胡颗粒的化学成分分析结果;筛选小柴胡颗粒的活性成分及其对应的作用靶点,构建药材‑活性成分‑作用靶点网络;筛选hub基因;基于作用靶点进行GO功能富集分析和KEGG信号通路富集分析,得到小柴胡颗粒的活性成分群。本发明筛选得到了33个活性成分组成了小柴胡颗粒的活性成分群,从整体角度系统探索了活性成分群与药效作用机制的关联。
Description
技术领域
本发明涉及中药研究领域,特别是涉及一种小柴胡颗粒活性成分群的构建方法。
背景技术
小柴胡颗粒药方源自名医张仲景《伤寒论》中经典名方“小柴胡汤”,由柴胡、黄芩、甘草、党参、姜半夏、生姜和大枣研制而成,目前广泛用于流感、发热、慢性肝炎、肝硬化和脂肪肝等疾病的治疗,是一个中医内科常用的有效方剂。目前小柴胡颗粒质量控制方面绝大多数集中于对黄芩苷成分的控制,监控指标成分单一,不能全面反映成品的质量信息。
中药的生物活性评价与其安全性和有效性相关,中药的药效物质基础是中药中与生物活性密切相关的化学成分群,因此,对中药中的生物活性成分的研究,阐明其药效物质基础是中药质量控制的一个重要步骤。目前,还缺少关于小柴胡颗粒的关于活性成分群构建的研究报道。
发明内容
基于此,本发明的目的是提供一种小柴胡颗粒活性成分群的构建方法,通过构建小柴胡颗粒的活性成分群从而从整体角度系统探索了活性成分群与药效作用机制的关联。
具体技术方案如下:
一种小柴胡颗粒活性成分群的构建方法,包括以下步骤:
取小柴胡颗粒浸膏样品,以甲醇水溶液为溶剂,制备供试品溶液;
取所述供试品溶液进行液相色谱质谱法分析,得到小柴胡颗粒的化学成分分析结果;
基于所得化学成分分析结果(全成分分析确证和指认的化合物),利用中药系统药理学数据库与分析平台或SwissADME数据库中的至少一种数据库筛选小柴胡颗粒的活性成分及其对应的作用靶点,并搜索所得作用靶点对应的标准基因名;根据小柴胡颗粒的药材、活性成分和作用靶点基因名通过Cytoscape数据库构建药材-活性成分-作用靶点网络;
基于所述作用靶点构建小柴胡颗粒靶蛋白互作网络,然后可视化小柴胡颗粒靶蛋白互作网络,筛选hub基因;
基于所述作用靶点进行GO功能富集分析和KEGG信号通路富集分析,得到小柴胡颗粒的活性成分群。
在其中一些实施例中,液相色谱的流动相A为乙腈,流动相B为0.05~0.15wt%甲酸水溶液,进行梯度洗脱。
在其中一些实施例中,所述梯度洗脱程序为:
0~30min:流动相A的体积百分比由(5±2)%上升到(50±2)%;
30~40min:流动相A的体积百分比由(50±2)%上升到(95±2)%;
40-44min:流动相A的体积百分比为(95±2)%;
44-45min:流动相A的体积百分比由(95±2)%降低至(5±2)%;
45-50min:流动相A的体积百分比为(5±2)%。
在其中一些实施例中,液相色谱的色谱条件还包括:流速:(0.3±0.05)ml/min;进样量:(3±0.5)μl;柱温:(30±3)℃。
在其中一些实施例中,液相色谱的色谱柱为Phenomenex kinetex C18。
在其中一些实施例中,质谱的色谱条件包括:电喷雾离子源,离子喷雾电压正模式(5500±50)V,离子喷雾电压负模式-(4500±50)V;喷雾气(55±5)psi;辅助加热气(55±5)psi;离子源温度(550±10)℃;气帘气(35±2)psi;碰撞气压力(10±1)psi;分别采用正、负离子模式进行检测。
在其中一些实施例中,所述供试品溶液的制备包括以下步骤:取小柴胡颗粒浸膏样品,加入60~80v/v%的甲醇水溶液,超声,过滤,取续滤液作为供试品溶液。
在其中一些实施例中,将所述作用靶点导入string数据库构建所述小柴胡颗粒靶蛋白互作网络。
在其中一些实施例中,应用Cytoscape可视化所述小柴胡颗粒靶蛋白互作网络。
在其中一些实施例中,将所述作用靶点导入DAVID数据库,进行GO功能富集分析和KEGG信号通路富集分析,得到小柴胡颗粒的活性成分群。
在其中一些实施例中,其中利用中药系统药理学数据库与分析平台筛选活性成分及其对应的靶点包括:基于所得化学成分分析结果,利用中药系统药理学数据库与分析平台,以口服生物利用度≥30、化合物类药性≥0.18为筛选条件,筛选活性成分及其对应的作用靶点。
在其中一些实施例中,其中利用SwissADME数据库筛选小柴胡颗粒的活性成分及其对应的作用靶点包括:基于所得化学成分分析结果,利用SwissADME数据库,以化合物的类药性参数达两个“Yes”以上为原则纳入活性成分群,并在SwissTarget Prediction数据库找到活性成分对应的作用靶点。
在其中一些实施例中,所述搜索所得作用靶点对应的标准基因名包括:通过UniProt数据库和Drugbank数据库中的至少一种数据库搜索所得作用靶点对应的标准基因名。
在其中一些实施例中,筛选hub基因包括:将所述作用靶点导入string数据库构建小柴胡颗粒靶蛋白互作网络,将蛋白种类设置为“人类”,最低相互作用得分设为中等置信度,将所得分析数据导入Cytoscape进行可视化分析,采用“cytoHubba”功能的“最大集团中心性”算法分析,选择得分前10的基因作为hub基因。
与现有技术相比,本发明具有以下有益效果:
本发明提供了一种柴胡颗粒活性成分群的构建方法,筛选了小柴胡颗粒的33个活性成分组成活性成分群,从整体角度系统探索了活性成分群与药效作用机制的关联。
附图说明
图1为小柴胡颗粒浸膏正模式(A)及负模式(B)总离子流图;
图2为小柴胡颗粒药材—成分—靶点网络图(蓝色代表药材,黄色代表靶点,红色代表活性成分,绿色代表共有成分,节点的大小代表度值(degree)大小CH:蓝色代表药材,黄色代表靶点,红色代表活性成分,绿色代表共有成分,节点的大小代表度值(degree)大小);
图3为小柴胡颗粒靶点相互作用网络(PPI)图;
图4为靶点相互作用网络中Hub基因;
图5为小柴胡颗粒核心靶点GO功能富集分析结果;
图6为小柴胡颗粒核心靶点KEGG通路富集分析结果。
具体实施方式
本发明下列实施例中未注明具体条件的实验方法,通常按照常规条件,或按照制造厂商所建议的条件。实施例中所用到的各种常用化学试剂,均为市售产品。
除非另有定义,本发明所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。本发明的说明书中所使用的术语只是为了描述具体的实施例的目的,不用于限制本发明。
本发明的术语“包括”和“具有”以及它们任何变形,意图在于覆盖不排他的包含。例如包含了一系列步骤的过程、方法、装置、产品或设备没有限定于已列出的步骤或模块,而是可选地还包括没有列出的步骤,或可选地还包括对于这些过程、方法、产品或设备固有的其它步骤。
在本发明中提及的“多个”是指两个或两个以上。“和/或”,描述关联对象的关联关系,表示可以存在三种关系,例如,A和/或B,可以表示:单独存在A,同时存在A和B,单独存在B这三种情况。字符“/”一般表示前后关联对象是一种“或”的关系。
以下结合具体实施例对本发明作进一步详细的说明。
实施例1小柴胡颗粒全成分分析
1实验材料
1.1仪器
十万分之一电子分析天平(MS105DU,瑞士Mettler toledo公司);万分之一电子分析天平(ME204,瑞士Mettler toledo公司);超纯水器(arium mini,德国Sartorius公司);数控超声波清洗器(KQ500DE,昆山市超声仪器有限公司);超快速高效液相色谱仪(LC-20AD-XR二元泵,SIL-20AD-XR自动进样器,CTO-20A柱温箱,SPD-M20A PDA检测器,日本岛津公司);四级杆-飞行时间质谱仪(Triple TOF 5600plus,美国AB SCIEX公司);色谱柱型号:Phenomenex kinetex C18色谱柱(3.0mm×150mm,2.6μm,S/N:H20-057369)。
1.2对照品
本研究实验所用对照品见表1-1。
表1-1对照品列表
1.3试剂
实验溶剂:甲醇(广州化学试剂厂,20180801,分析纯)。
液相色谱所用溶剂:甲醇、乙腈(Honeywell,Q8AG1H,色谱级),甲酸(F&H,2017011811A,液质级),Sartorius超纯水。
1.4供试品
小柴胡颗粒浸膏(广州白云山光华制药股份有限公司,20200618)。
2实验方法
2.1对照品溶液的制备
分别称取黄芩苷、汉黄芩苷、黄芩素、汉黄芩素、甘草苷、芹菜素、槲皮素、山柰酚、山柰酚-3-O-芸香糖苷、柴胡皂苷A、柴胡皂苷D、柴胡皂苷B1、柴胡皂苷B2、甘草酸、绿原酸、阿魏酸、异阿魏酸、精氨酸、芦丁等对照品适量,精密称定,加甲醇制成1mL各含10μg的混合对照品溶液。
2.2供试品溶液的制备
称取小柴胡颗粒浸膏约50mg,精密称定,置具塞锥形瓶中,加入70%甲醇溶液到25ml容量瓶,超声处理(功率300w,频率40kHz)30分钟,放冷,再用70%甲醇定容,摇匀,用0.22μm微孔滤膜滤过,取续滤液1ml,加入内标工作液10ul,混匀即得。
2.3检测条件
液相色谱条件:以Phenomenex kinetex C18(3.0×150mm,2.6μm)为色谱柱;柱温:30℃;流动相:乙腈(A)-0.1%甲酸(B);梯度洗脱程序为:0~30min:5%-50%A;30~40min:50%-95%A;40~44min:95%A,44~45min:95%-5%A;45~50min:5%A;流速:0.3ml/min;进样量:3μl。
质谱条件:ESI电喷雾离子源,TOF MS扫描质量范围为m/z 100-1500,TOF MS/MS扫描质量范围为m/z 50-1500。离子喷雾电压正模式5500V,负模式-4500V;喷雾气55psi;辅助加热气55psi;离子源温度550℃;气帘气35psi;碰撞气压力10psi,分别采用正、负离子模式进行检测。
3数据处理
应用PeakviewTM2.1(美国AB Sciex公司)对数据进行分析。通过对照品对照、谱库检索(中药质谱数据库,1.0版本,美国AB Sciex公司)、准确分子量、裂解碎片及文献,对样品中的化合物进行指认或确证。
4结果与讨论
质谱检测采用正负模式进行一级和二级扫描,小柴胡颗粒浸膏供试品溶液的正负模式总离子流图见图1。通过对照品对照、准确分子量比较、裂解碎片分析和谱库检索匹配,确证和指认到101个化合物,包括57种黄酮类、4种生物碱、8种氨基酸类、17种萜类、5种糖醇类及10种有机酸类成分。各化合物在正、负模式下的保留时间、峰归属、分子离子峰及详细裂解碎片信息见表1-2。采用对照品对照未检测到柴胡皂苷D、异阿魏酸、金丝桃苷、秦皮乙素、白术内酯III、山柰酚-3-O-芸香糖苷和芹菜素。
表1-2小柴胡颗粒浸膏化学成分分析
备注:a对照品确证.b碎片丢失:Glc-葡萄糖,Rha-鼠李糖,ND-表示没有检测到响应.c谱库检索对照(中药质谱数据库,1.0版本,美国AB Sciex公司).
a Confirmation in comparison with authentic standards.b The lossesare:Glc=glucose moiety,Rha=rhamnose moiety,ND=not detect.c Confirmation incomparison with mass spectral library(Natural Products HR-MS/MS SpectralLibrary,Version 1.0;AB Sciex,Foster City,USA).
实施例2小柴胡颗粒活性成分群的构建
1方法
1.1药材-活性成分-作用靶点网络的构建
基于小柴胡颗粒全成分分析确证和指认的101个化合物,利用中药系统药理数据库(TCMSP)(http://tcmspw.com/tcmsp.php)分析平台,以口服生物利用度(oralbioavailability,OB)≥30%、化合物类药性(drug-like,DL)≥0.18为筛选条件,筛选活性化合物及对应靶点。对于无法在该平台搜索的化学成分,经SwissADME(http://www.swissadme.ch/)数据库,以Druglikeness参数达两个“Yes”以上为原则纳入活性成分群并在SwissTargetPrediction(http://www.swisstargetprediction.ch/)找到对应靶点(Probability>0)。通过UniProt(https://www.uniprot.org/)、Drugbank(https://www.drugbank.ca/)等数据库搜索靶蛋白对应的标准基因名,限定物种为“Homo sapiens(人类)”。将药材、活性成分与靶点相应基因导入Cytoscape 3.7.0(https://cytoscape.org//),构建可视化的药材—活性成分—作用靶点网络,并分析网络的拓扑特征。
1.2靶蛋白相互作用(PPI)网络构建及hub基因筛选
使用String平台(https://string-db.org/)构建小柴胡颗粒靶蛋白互作(PPI)网络,将蛋白种类设置为“Homo sapiens(人类)”进行操作,最低相互作用得分设为中等置信度“medium confidence(0.4)”,其余保持默认设置。将分析数据导入Cytoscape3.7.0,进行网络可视化分析。采用“cytoHubba”功能的“最大集团中心性(maximal cliquecentrality,MCC)”算法分析,选择得分前10的基因作为hub基因。
GO功能富集、KEGG通路分析
将靶点基因导入DAVID v6.7数据库(https://david.ncifcrf.gov/)并限定物种为人(Homo sapiens),进行GO功能富集分析和KEGG(Kyoto Encyclopedia of Genes andGenomes)信号通路富集分析。
2结果
2.1活性化学成分筛选及其靶点的收集
利用TCMSP和SwissADME数据库筛选得到小柴胡颗粒活性成分共33个(见表2-1),汇总收集对应靶点共690个。
表2-1小柴胡颗粒主要活性成分
备注:CH:柴胡;HQ:黄芩;BX:姜半夏;DS:党参;GC:甘草;SJ:生姜;DZ:大枣。
2.2药材—成分—靶点网络分析
经Cytoscape 3.7.2软件分析建立的药材—成分—靶点网络见图2。由图2可知,此网络共有730个节点(7味药材,33个成分,690个靶点),2169条边,其中蓝色代表药材,黄色代表靶点,红色代表活性成分,绿色代表共有成分,结合“Network Analysis”分析,节点的大小代表度值(degree)大小,以上33个成分的度值(degree)均超过10,各成分对应的靶点视为潜在作用靶点。degree值前5的成分是C1:槲皮素、A1:黄芩素、HQ2:汉黄芩素、B1:异鼠李素和GC3:光甘草定。
2.3靶蛋白相互作用(PPI)网络构建及hub基因筛选
将690个靶点导入String数据库构建PPI网络,隐藏独立无连接的蛋白(见图3)。应用Cytoscape可视化PPI网络并计算Hub基因。图4所示为排名前10的Hub基因,分别是VEGFA、JUN、MAPK3、MMP9、IL6、PTGS2、CXCL8、TP53、GAPDH、STAT3,结果提示这些靶点可能为小柴胡颗粒发挥作用的关键靶点。
2.4 GO功能富集、KEGG通路分析结果
运用DAVID平台的GO富集分析和KEGG通路分析的功能,对小柴胡颗粒中的690个核心靶点进行研究。通过GO富集分析确定了550个GO条目(P<0.05),提示小柴胡颗粒主要参与的生物学过程包括生物对营养元素的应答过程(response to nutrient levels)、类固醇代谢进程(steroid metabolic process)、生长调控(regulation of growth)、肌细胞增殖(muscle cell proliferation)、对脂多糖的应答(response to lipopolysaccharide)、细胞对脂质的应答(cellular response to lipid)。图5列举的分别为GO功能富集到的细胞、分子及生物学功能排名前十的条目,其中细胞组成相关条目主要涉及细胞质、细胞膜、细胞膜组分;分子功能相关条目主要涉及酶结合、蛋白激酶活性、药物绑定、ATP结合等;提示小柴胡颗粒主要参与的生物学过程包括生物对药物的应答过程(response to drug)、对酒精的应答、对凋亡过程的负调控、蛋白质磷酸化等。通过KEGG通路分析共筛选到142条通路(筛选FDR<0.05的通路,FDR值越小,富集程度越高)。图6为列举的前20条相关通路,主要包括免疫与炎症免疫调控相关的Metabolic pathways、Rap1signaling pathway、Neuroactiveligand-receptor interaction、MAPK pathway、PI3K-Akt signaling pathway、cAMPsignaling pathway;抗肿瘤相关通路Pathways in cancer、Proteoglycans in cancer;与病毒性疾病相关的Hepatitis B、Viral carcinogenesis,influenza A;与肺纤维化调控相关的PI3K-Akt signaling pathway、HIF-1signaling pathway和FoxO signaling pathway等。
综合以上,本发明基于UFLC-Q-TOF-MS/MS技术确证和指认的小柴胡颗粒101个化学成分,运用TCMSP、Cytoscape、String和DAVID等平台筛选了小柴胡颗粒的33个活性成分组成活性成分群,并从整体角度系统探索了活性成分群与药效作用机制的关联。
以上所述实施例的各技术特征可以进行任意的组合,为使描述简洁,未对上述实施例中的各个技术特征所有可能的组合都进行描述,然而,只要这些技术特征的组合不存在矛盾,都应当认为是本说明书记载的范围。
以上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
Claims (10)
1.一种小柴胡颗粒活性成分群的构建方法,其特征在于,包括以下步骤:
取小柴胡颗粒浸膏样品,以甲醇水溶液为溶剂,制备供试品溶液;
取所述供试品溶液进行液相色谱质谱法分析,得到小柴胡颗粒的化学成分分析结果;
基于所得化学成分分析结果,利用中药系统药理学数据库与分析平台或SwissADME数据库中的至少一种数据库筛选小柴胡颗粒的活性成分及其对应的作用靶点,并搜索所得作用靶点对应的标准基因名;根据小柴胡颗粒的药材、活性成分和作用靶点基因名通过Cytoscape数据库构建药材-活性成分-作用靶点网络;
基于所述作用靶点构建小柴胡颗粒靶蛋白互作网络,然后可视化小柴胡颗粒靶蛋白互作网络,筛选hub基因;
基于所述作用靶点进行GO功能富集分析和KEGG信号通路富集分析,得到小柴胡颗粒的活性成分群。
2.根据权利要求1所述的构建方法,其特征在于,液相色谱的流动相A为乙腈,流动相B为0.05~0.15wt%甲酸水溶液,进行梯度洗脱。
3.根据权利要求2所述的构建方法,其特征在于,所述梯度洗脱程序为:
0~30min:流动相A的体积百分比由(5±2)%上升到(50±2)%;
30~40min:流动相A的体积百分比由(50±2)%上升到(95±2)%;
40-44min:流动相A的体积百分比为(95±2)%;
44-45min:流动相A的体积百分比由(95±2)%降低至(5±2)%;
45-50min:流动相A的体积百分比为(5±2)%。
4.根据权利要求1所述的构建方法,其特征在于,液相色谱的色谱条件还包括:
流速:(0.3±0.05)ml/min;进样量:(3±0.5)μl;柱温:(30±3)℃;色谱柱为Phenomenex kinetex C18。
5.根据权利要求1所述的构建方法,其特征在于,质谱的色谱条件包括:电喷雾离子源,离子喷雾电压正模式(5500±50)V,离子喷雾电压负模式-(4500±50)V;喷雾气(55±5)psi;辅助加热气(55±5)psi;离子源温度(550±10)℃;气帘气(35±2)psi;碰撞气压力(10±1)psi;分别采用正、负离子模式进行检测。
6.根据权利要求1~5任一项所述的构建方法,其特征在于,所述供试品溶液的制备包括以下步骤:取小柴胡颗粒浸膏样品,加入60~80v/v%的甲醇水溶液,超声,过滤,取续滤液作为供试品溶液。
7.根据权利要求1~5任一项所述的构建方法,其特征在于,将所述作用靶点导入string数据库构建所述小柴胡颗粒靶蛋白互作网络;
和/或,应用Cytoscape可视化所述小柴胡颗粒靶蛋白互作网络;
和/或,将所述作用靶点导入DAVID数据库,进行GO功能富集分析和KEGG信号通路富集分析,得到小柴胡颗粒的活性成分群。
8.根据权利要求1~5任一项所述的构建方法,其特征在于,其中利用中药系统药理学数据库与分析平台筛选活性成分及其对应的靶点包括:基于所得化学成分分析结果,利用中药系统药理学数据库与分析平台,以口服生物利用度≥30、化合物类药性≥0.18为筛选条件,筛选活性成分及其对应的作用靶点。
9.根据权利要求1~5任一项所述的构建方法,其特征在于,其中利用SwissADME数据库筛选小柴胡颗粒的活性成分及其对应的作用靶点包括:基于所得化学成分分析结果,利用SwissADME数据库,以化合物的类药性参数达两个“Yes”以上为原则纳入活性成分群,并在SwissTarget Prediction数据库找到活性成分对应的作用靶点;
和/或,所述搜索所得作用靶点对应的标准基因名包括:通过UniProt数据库和Drugbank数据库中的至少一种数据库搜索所得作用靶点对应的标准基因名。
10.根据权利要求1~5任一项所述的构建方法,其特征在于,筛选hub基因包括:将所述作用靶点导入string数据库构建小柴胡颗粒靶蛋白互作网络,将蛋白种类设置为“人类”,最低相互作用得分设为中等置信度,将所得分析数据导入Cytoscape进行可视化分析,采用“cytoHubba”功能的“最大集团中心性”算法分析,选择得分前10的基因作为hub基因。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105486771A (zh) * | 2015-12-23 | 2016-04-13 | 成都普思生物科技股份有限公司 | 小柴胡颗粒复方制剂的指纹图谱检测方法 |
CN110201130A (zh) * | 2019-03-06 | 2019-09-06 | 十堰市太和医院 | 一种小柴胡汤提取物的制备及质谱指纹图谱构建方法 |
CN110320286A (zh) * | 2018-03-29 | 2019-10-11 | 广州白云山光华制药股份有限公司 | 小柴胡颗粒有效成分的含量测定方法 |
CN110970116A (zh) * | 2019-12-05 | 2020-04-07 | 吉林省蒲川生物医药有限公司 | 一种基于转录组学的中药药理机制分析方法 |
-
2021
- 2021-04-20 CN CN202110426765.1A patent/CN113156011A/zh active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105486771A (zh) * | 2015-12-23 | 2016-04-13 | 成都普思生物科技股份有限公司 | 小柴胡颗粒复方制剂的指纹图谱检测方法 |
CN110320286A (zh) * | 2018-03-29 | 2019-10-11 | 广州白云山光华制药股份有限公司 | 小柴胡颗粒有效成分的含量测定方法 |
CN110201130A (zh) * | 2019-03-06 | 2019-09-06 | 十堰市太和医院 | 一种小柴胡汤提取物的制备及质谱指纹图谱构建方法 |
CN110970116A (zh) * | 2019-12-05 | 2020-04-07 | 吉林省蒲川生物医药有限公司 | 一种基于转录组学的中药药理机制分析方法 |
Non-Patent Citations (7)
Title |
---|
LIN HAN等: "Potential mechanism prediction of Cold-Damp Plague Formula against COVID-19 via network pharmacology analysis and molecular docking", 《CHIN MED》 * |
刘晓帆等: "采用HPLC-TOF/MS对中药复方小柴胡汤中化学成分的快速分析鉴别", 《第二军医大学学报》 * |
孙凯滨等: "小柴胡汤治疗早期新型冠状病毒肺炎(COVID-19)邪热郁肺、枢机不利证功效网络分析与机制预测", 《中草药》 * |
杨璐等: "基于网络药理学的小柴胡汤治疗新型冠状病毒肺炎(COVID-19)发热的可行性探讨", 《中草药》 * |
毕聪 等: "小柴胡颗粒生物靶标网络及潜在作用机制解析", 《中山大学学报(自然科学版)》 * |
毕聪等: "小柴胡颗粒基于网络药理学的组方配伍规律解析", 《药学研究》 * |
郑如文等: "小柴胡颗粒的高效液相指纹图谱研究", 《中国医院用药评价与分析》 * |
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