基于谱效关联分析和网络药理学研究连蒲双清散药效物质及作用靶点

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国家科技期刊平台
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中文摘要：[目的]基于试验设计构建连蒲双清散抗炎抑菌的谱效关联分析，并结合网络药理及体内外试验揭示其药效物质基础及作用机制。[方法]选取黄连蒲公英比例、提取酸浓度、乙醇浓度为影响因素针对性设计L9(34)正交试验，得到9种连蒲双清散提取物，采用高效液相色谱(HPLC)法建立连蒲双清散不同提取物的指纹图谱;以脂多糖(LPS)诱导的小鼠巨噬细胞炎症指标和对金黄色葡萄球菌的抑菌圈直径作为药效学评价指标，开展连蒲双清散的体外药效学研究;采用灰色关联度分析法及偏最小二乘回归分析法建立谱效关系，筛选出其发挥抗炎药效和对金黄色葡萄球菌有抑菌效果的药效物质。通过网络药理手段从另一个层面筛选药效成分和靶点，并通过体内试验对谱效分析和网络药理学的结果进行验证。[结果]HPLC指纹图谱识别出9个共有峰，并鉴定出4种组分(菊苣酸、黄连碱、表小檗碱和巴马汀)。正交试验结果显示，黄连中的成分对金黄色葡萄球菌的抑菌效果起主导作用，而蒲公英对抗炎药效的发挥作用更明显。谱效分析筛选出表小檗碱和黄连碱是连蒲双清散的部分药效物质基础。网络药理学挖掘出表小檗碱、巴马汀、黄连碱、木犀草素和咖啡酸是网络中权重最高的5个成分，白细胞介素-6(IL6)和肿瘤坏死因子(TNF)是其中2个主要的作用靶点;分子对接结果显示，5个化合物与2个靶点蛋白结合能均低于-5 kJ/mol，对接良好。小鼠急性肠炎模型中，病理切片显示，经连蒲双清散治疗后的小鼠结肠中炎症细胞浸润病变面积明显减少;ELISA检测结果显示，药物组与模型组小鼠结肠中炎症因子表达量具有显著差异(P＜0。05)。[结论]基于试验设计的谱效关联分析结合网络药理学有效挖掘出连蒲双清散发挥抗炎作用和对金黄色葡萄球菌有抑菌效果的主要药效物质是表小檗碱和黄连碱，主要作用靶点是IL6和TNF。本研究为探讨复方中药的药效物质基础及作用靶点提供了有意义的尝试。

外文标题：Study on the Pharmacodynamic Substances and Targets of Lianpu Shuangqing Powder Based on Spectrum-effect Relationship Analysis and Network Pharmacology

外文摘要：[Objective]The spectrum-effect correlation analysis of the anti-inflammatory and bacteriostatic properties of Lianpu Shuangqing powder was constructed based on experimental design,and its pharmacological substance basis and mechanism of action were revealed through the integration of network pharmacology and in vitro and in vivo experiments.[Method]The L9(34)orthogonal experiment was devised incorporating the proportion of Coptidis Rhizoma and Taraxaci herba,the extracted acid concentration and the ethanol concentration,resulting in nine pairs of extracts of Lianpu Shuangqing powder.HPLC was utilized to establish fingerprints for each extract.An in vitro pharmacodynamic analysis of anti-inflammatory and bacteriostatic effects was conducted utilising lipopolysaccharide(LPS)-induced inflammatory cytokines of mouse macrophages and bacteriostasis zone as pharmacodynamic evaluation indexes.The spectrum-effect relationship was established using the grey relational degree analysis and partial least squares regression analysis methods,which screened out the pharmacological substances responsible for its anti-inflammatory effects and bacteriostatic effects against Staphylococcus aureus.Pharmacodynamic constituents and targets were further identified through network pharmacology analysis.Lastly,in vivo experiment was used to verify the spectral and network pharmacology results.[Result]HPLC fingerprints identified nine common peaks and four components(chicoric acid,coptisine,epiberberine and palmatine).The results of orthogonal test showed that the components in Coptidis Rhizoma played a dominant role in the antibacterial effect against Staphylococcus aureus,while the role of Taraxaci herba in the anti-inflammatory efficacy was more obvious.Spectrum-effect analysis screened that epiberberine and coptisine were part of the pharmacological substance basis for the anti-inflammatory and antibacterial effects of the powder.Network pharmacology research revealed that epiberberine,palmatine,coptisine,luteolin and caffeic acid were the five components with the highest weights in the network.Interleukin-6(IL6)and tumor necrosis factor(TNF)were the two main anti-inflammatory and antibacterial targets.The docking results showed that the binding energies of five compounds to both target protein receptors were all below-5 kJ/mol,indicating a good binding affinity.In the mouse model of acute enteritis,histological sections revealed that the area of inflammatory cell infiltration in the colon of mice was significantly reduced after treatment with Lianpu Shuangqing powder.ELISA experiments showed a significant difference in the expression of inflammatory factors in colon of mice between the drug group and the model group(P＜0.05).[Conclusion]An integrated approach involving spectrum-effect correlation analysis based on experimental design and network pharmacology had effectively delineated that the primary pharmacological substances responsible for the anti-inflammatory effects and bacteriostatic effects against Staphylococcus aureus in Lianpu Shuangqing powder were coptisine and epiberberine,with IL6 and TNF being the major target molecules.This study offered a significant exploration into the pharmacological substance basis and target actions of compound traditional Chinese medicines.

外文关键词：

Lianpu Shuangqing powderexperimental designspectrum-effect relationshipanti-inflammatory and bacteriostaticpharmacodynamic substance

作者：

齐小雨、刘星彤、孙一丹、候冉冉、李秋、刘志海、刘聪敏

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作者单位：

青岛农业大学化学与药学院,青岛 266109

青岛农业大学动物医学院,青岛 266109

关键词：

连蒲双清散试验设计谱效关系抗炎抑菌药效物质

基金：

山东省自然科学基金项目国家自然科学基金项目山东省重点研发计划项目山东省重点研发计划项目青岛农业大学高层次人才科研启动基金项目中国博士后科学基金项目

项目编号：

ZR2020QC198820039222021CXGC0113052022LYXZ03066311210422021M701105

出版年：

2024

DOI：

10.16431/j.cnki.1671-7236.2024.08.032

中国畜牧兽医

中国农业科学院北京畜牧兽医研究所

中国畜牧兽医

CSTPCD北大核心

影响因子：0.72

ISSN：1671-7236

年,卷(期)：2024.51(8)