益肾排石方的指纹图谱及功效关联物质的作用机制

Fingerprints and Action Mechanism of Efficacy-Associated Substances of Yishen Paishi Formula

应用液质联用技术分析了益肾排石方中的化学成分,建立高效液相色谱法(HPLC)指纹图谱,结合生物信息学技术对关键成分、关键靶点和通路进行了预测分析。应用超高效液相串联质谱技术(UHPLC-MS/MS)对益肾排石方中的化学成分进行了鉴定。10批益肾排石方样品,应用HPLC建立指纹图谱,标记共有峰,进行共有峰归属,并进行相似度评价。应用生物信息学技术,基于指认出的成分,建立了“中药-成分-靶点”网络图,通过GEO数据库获得了肾结石相关的差异基因。对方中功效关联物质的作用机制进行了进一步的分析,并进行了分子对接验证。通过数据库匹配、元素组成和碎片结构分析,共从益肾排石方中鉴定出32种成分;在对10批益肾排石方的指纹图谱的建立过程中,通过对照品指认确定了王不留行黄酮苷、松脂醇二葡萄糖苷、虎杖苷、黄芩苷、黄芩素、汉黄芩苷、芦丁、木蝴蝶苷A、大黄酸、异橙黄酮、芦荟大黄素、白杨素、大黄酚、厚朴酚、大黄素甲醚、汉黄芩素和大黄素17个共有峰,10批益肾排石方相似度结果均在合理范围内,且17个共有峰均能归属到虎杖、黄芩、牛膝、柴胡、金钱草、枳实、大黄、厚朴、杜仲和王不留行10味中药。结合生物信息学筛选了ESR1和PTGS1为关键基因,与17种药效成分进行分子对接验证,预测大黄酸和王不留行黄酮苷为关键成分。

The chemical components in Yishen Paishi Formula are analyzed by applying liquid chromatographymass spectrometry technique,and high-performance liquid chromatography(HPLC)fingerprints are established,combined with bioinformatics techniques to predict the key components,key targets and pathways.The chemical components in Yishen Paishi Formula are identified by ultrahigh-performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS),and fingerprint profiles are established by HPLC.Similarity evaluation is performed on 10 batches of Yishen Paishi formula,and shared peaks are labeled for shared peak attribution.Based on the recognized components,a“traditional Chinese medicine-ingredient-target”network diagram is established using bioinformatics,and the differential genes related to kidney stones are obtained from the GEO database.The mechanism of action of the efficacy-associated substances in the formula is further analyzed and molecular docking verification is performed.Results Through database matching,elemental composition and fragment structure analysis,a total of 32 components are identified from the Yishen Paishi formulae;in the process of establishing the fingerprint profiles of 10 batches of Yishen Paishi formulae,17 common peaks are identified through the fingerprinting of controls,including Vaccarin,pinoresinol diglucoside,polydatin,Baicalin,baicalein,Wogonoside,Rutinum,Oroxin A,rhein,Isosinensetin,aloe-emodin,chrysin,Chrysophanol,Magnolol,physcion,Wogonin and emodin.The similarity results of the 10 batches of Yishen Paishi formula are all within reasonable range,and all 17 peaks could be attributed to 10 traditional Chinese medicines,such as Polygoni Cuspidati Rhizomaer Radix,Scutellariae Radix,Achyranthis Bidentatae Radix,Bupleuri Radix,Lysimachiae Herba,Aurantii Fructus Immaturus,Rhei Radix Et Rhizoma,Magnolia Officinalis Cortex,Eucommiae Cortex and Vaccariae Semen.Combined with bioinformatics,ESR1 and PTGS1 are screened as key genes and validated by molecular docking with 17 potent components,and rhein and Vaccarin are predicted as key components.