基于HPLC和网络药理学方法的三化汤治疗脑缺血活性成分含量测定及作用靶点预测  

Determination of active ingredients and prediction of targetin Sanhua Decoction for cerebral ischemia based on HPLC and network pharmacology methods

作　　者：路颖慧 黄颖[2] 孙明杰[2] 李雯洁 Lu Yinghui;Huang Ying;Sun Mingjie;Li Wenjie(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Medical Experiment Center,China Academy of Chinese Medical Sciences,Beijing Key Laboratory of Research of Chinese Medicine on Prevention and Treatment for Major Diseases,Beijing 100700,China)

机构地区：[1]北京中医药大学中药学院,102488 [2]中国中医科学院医学实验中心,中医药防治重大疾病基础研究北京市重点实验室,北京100700

出　　处：《国际中医中药杂志》2021年第11期1109-1115,共7页International Journal of Traditional Chinese Medicine

基　　金：北京市自然科学基金(7192142);中央级公益性科研院所基本科研业务费专项资金(ZZ2018005)。

摘　　要：目的:结合HPLC和网络药理学方法对三化汤治疗脑缺血的活性成分进行含量测定,预测其发挥作用的潜在靶点。方法:采用HC-C18色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇-乙腈-0.05%磷酸水,梯度洗脱,流速1.0、0.8 ml/min;检测波长254 nm,柱温30℃,进样量10μl。运用TCMIP V 2.0平台检索大黄酸、大黄素、大黄酚、橙皮苷、厚朴酚和羌活醇6种成分作用靶点,以Cerebral ischemia为关键词,检索GeneCards、在线人类孟德尔遗传数据库(OMIM)、疗效药靶数据库(TTD)、Disgenet数据库,筛选6种活性成分治疗脑缺血潜在靶点,构建蛋白质相互作用网络和疾病-成分-靶点-通路整合网络。结果:三化汤中6种活性成分大黄酸、大黄素、大黄酚、橙皮苷、厚朴酚和羌活醇线性范围分别为0.0804~0.8040μg、0.0153~0.3820μg、0.0418~0.6264μg、0.3126~3.9080μg、0.0379~0.5688μg、0.0453~1.3596μg,相关系数均大于0.9990。经测定7份样品中上述6个成分含量范围分别为0.887~0.971 mg/g、0.094~0.101 mg/g、0.110~0.119 mg/g、1.494~1.669 mg/g、0.126~0.145 mg/g、0.153~0.167 mg/g。网络药理学分析发现,TNF、TP53、MAPK14、JUN、IL1B、MYC、ESR1、ICAM1、PTGS2、PPARG为三化汤中6种活性成分治疗脑缺血的核心靶点。结论:建立了三化汤治疗脑缺血6种活性成分质量控制方法,该方法样品处理简便,色谱条件简单,结果准确。明确了三化汤中6种活性成分治疗脑缺血的10个潜在靶点,为进一步研究其作用机制奠定基础。Objective:To determinate the active ingredients and predicte action targets in Sanhua Decoction for treating cerebral ischemia based on HPLC and network pharmacology methods.Methods:The HPLC analysis was performed on HC-C18(250 mm×4.6 mm,5μm)with the mobile phase of methanolacetonitrile-0.05%phosphoric acid with gradient elution at the flow rate of 1.0 ml/min and 0.8 ml/min.The detection wavelength was set at 254 nm,the column temperature was at 30℃,and the injection volume was 10μl.TCMIP v2.0 platform was used to search the action targets of rhein,emodin,chrysophanol,hesperidin,magnolol and notopterygiol.We used Cerebral ischemia as a keyword searching the databases such as Genecards,OMIM,TTD,and Disgenet to screen six potential targets for the treatment of cerebral ischemia by active ingredients,construct a protein interaction(PPI)network and a disease-component-target-pathway integration network.Results:The literature search determined that rhein,emodin,chrysophanol,hesperidin,magnolol and qianhuol were the index components for the determination of Sanhua Decoction.Their linear ranges were 0.0804-0.8040μg,0.0153-0.3820μg,0.0418-0.6264μg,0.3126-3.9080μg,0.0379-0.5688μg,0.0453-1.3596μg,respectively.The correlation coefficient R 2 is greater than 0.9990.The content ranges of the above six components in seven samples were 0.887-0.971 mg/g,0.094-0.101 mg/g,0.110-0.119 mg/g,1.494-1.669 mg/g,0.126-0.145 mg/g and 0.153-0.167 mg/g,respectively.Network pharmacology analysis found that the targets of the six components for the treatment of cerebral ischemia may be TNF,TP53,MAPK14,JUN,IL1B,MYC,ESR1,ICAM1,PTGS2,PPARG and so on.Conclusions:A quality control method forthe six active ingredients in Sanhua Decoction treating cerebral ischemia was established.This method is simple and repeatable.The ten potential targets of the six active ingredients in Sanhua Decoction for the treatment of cerebral ischemia have been clarified,laying a foundation for further research on the action mechanism.

关 键 词：三化汤 脑缺血 网络药理学 靶点 含量测定 质量控制 

分 类 号：R285[医药卫生—中药学]