目的 基于HPLC指纹图谱、化学模式识别、网络药理学与分子对接，筛选能够表征3个基原文蛤肉共性药效物质的潜在质量标志物（quality markers，Q-Marker），建立多基原共用的含量测定方法，为多基原文蛤肉药材统一质量控制提供依据。方法 采集10批文蛤、14批琴文蛤、8批青蛤共32批样品，建立HPLC氨基酸指纹图谱，开展相似度评价与聚类分析（cluster analysis，CA）、主成分分析（principal component analysis，PCA）、正交偏最小二乘判别分析（orthogonal partial least squares discriminant analysis，OPLS-DA），解析不同基原整体化学轮廓的相似性与差异性；选取3个基原共有的氨基酸成分，采用网络药理学构建“成分-靶点-通路”网络，结合分子对接预测成分-靶点结合活性；对筛选得到的潜在Q-Marker开展方法学验证与多基原样品定量测定。结果 文蛤、琴文蛤、青蛤分别标定13、13、11个共有峰，指认谷氨酸、甘氨酸、牛磺酸、精氨酸、丙氨酸为3个基原共有的特征氨基酸；不同基原间相似度≥0.862，整体化学轮廓相似度较高，但化学模式识别可实现3个基原的区分，提示基原间客观存在成分含量差异。网络药理学筛选得到GOT2、GATM等9个核心靶点，主要涉及内分泌、免疫、神经相关信号通路，提示可能参与抗肿瘤、抗氧化、神经保护、能量代谢调节等相关生物学过程；分子对接结果显示，5个共有氨基酸与核心靶点具有一定结合亲和力，但整体结合强度偏弱至中等。建立5种氨基酸HPLC定量方法，方法学可靠；32批样品中5种氨基酸均可检出，但不同基原成分含量存在明显分布特征。结论 尽管文蛤肉3个基原在部分成分含量上存在差异，但谷氨酸、甘氨酸、牛磺酸、精氨酸、丙氨酸为3个基原共有、可测且潜在药效相关的物质，适合作为多基原文蛤肉共用潜在Q-Marker；可为多基原海洋中药材“同药多源”的整体质量评价提供研究范例。

Objective Based on HPLC fingerprint, chemical pattern recognition, network pharmacology and molecular docking, potential quality markers (Q-Markers) characterizing the common chemical components of three origins of Meretricis seu cyclinae musculus were screened, and a content determination method for multi-origin M. seu cyclinae musculus was established, providing a basis for unified quality control of multi-origin M. seu cyclinae musculus. Methods A total of 32 batches of samples, including ten batches of Meretrix meretrix Linnaeus, 14 batches of Meretrix lyrata Sowerby, and eight batches of Cyclina sinensis Gmelin, were collected to establish HPLC amino acid fingerprints. Similarity evaluation and cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were carried out to analyze the similarity and difference of the overall chemical profiles among different origins. The amino acid components shared by the three origins were selected, and the component-target-pathway network was constructed by network pharmacology, and the component-target binding activity was predicted by molecular docking. Methodological validation and quantitative determination of multi-origin samples were performed for the selected potential Q-Markers. Results A total of 13, 13, and 11 common peaks were identified in M. meretrix, M. lyrata, and C. sinensis, respectively. Glutamic acid, glycine, taurine, arginine, and alanine were identified as the characteristic amino acids shared by the three origins. The similarity among different origins was ≥ 0.862, indicating a high similarity in the overall chemical profile. However, chemical pattern recognition could distinguish the three origins, suggesting differences in component content among them. Nine core targets such as GOT2 and GATM were screened by network pharmacology, mainly involving endocrine, immune, and nerve-related signaling pathways, suggesting that these pathways may be involved in anti-tumor, anti-oxidation, neuroprotection, energy metabolism regulation, and other related biological processes. Molecular docking results suggested that the five common amino acids showed certain binding affinity with the core targets, but the overall binding strength was weak to moderate. HPLC quantitative methods for the five amino acids were established, and the methodology was reliable. Five amino acids could be detected in 32 batches of samples, but the contents of components from different origins showed obvious distribution characteristics. Conclusion Although there were differences in the content of some components among the three origins of M. seu cyclinae musculus, glutamic acid, glycine, taurine, arginine, and alanine were common, measurable, and potentially bioactive substances in the three origins, and are suitable as potential quality markers for multi-origin M. seu cyclinae musculus. This study provides a research example for the overall quality evaluation of multi-origin marine Chinese medicinal materials with “"multi-origin of the same drug”.

R282.6

广西中药药效研究重点实验室项目（桂科计字〔2005〕6号）；广西高校工程研究中心—“中药资源循环利用广西高校工程研究中心”