目的 基于超分子“印跡模板”理論，首創(chuàng)因子旋轉(zhuǎn)法與分子連接性指數(shù)（molecular connectivity index，MCI）耦合策略，系統(tǒng)解析活血化瘀中藥的共性物質(zhì)基礎(chǔ)及其“結(jié)構(gòu)-功能”關(guān)聯(lián)機(jī)制，為中藥質(zhì)量標(biāo)志物（quality markers，Q-Marker）篩選提供新范式。方法 采用UPLC法建立13味活血化瘀中藥（川芎、丹參、桃仁、馬鞭草、紅花、延胡索、月季花、牛膝、益母草、三七、白芍、赤芍、當(dāng)歸）及3種經(jīng)典復(fù)方（補(bǔ)陽(yáng)還五湯、桃紅四物湯、血府逐瘀湯）的指紋圖譜，通過(guò)匹配頻數(shù)法劃分26個(gè)“印跡模板”成分簇作為結(jié)構(gòu)性“物質(zhì)單元”；利用因子旋轉(zhuǎn)法對(duì)成分簇進(jìn)行降維整合，提取6個(gè)功能性“物質(zhì)單元”（累積方差貢獻(xiàn)率88.37%），結(jié)合歸一化評(píng)分評(píng)估給藥性“物質(zhì)單元”的綜合效應(yīng)；借助中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫(kù)與分析平臺(tái)（Traditional Chinese Medicine Systems Pharmacology Database，TCMSP）數(shù)據(jù)庫(kù)計(jì)算MCI，量化成分簇結(jié)構(gòu)相似性，并通過(guò)急性血瘀大鼠模型驗(yàn)證核心成分的藥效貢獻(xiàn)。結(jié)果 發(fā)現(xiàn)活血化瘀中藥的“印跡模板”成分簇可劃分為6個(gè)功能性“物質(zhì)單元”，其中前3個(gè)功能性“物質(zhì)單元”（累積貢獻(xiàn)率66.85%）以黃酮類、生物堿類及萜類為核心，主導(dǎo)活血化瘀效應(yīng)；補(bǔ)陽(yáng)還五湯因黃酮類、生物堿類及萜類的高豐度匹配，綜合評(píng)分（2.44）顯著優(yōu)于其他方劑，證實(shí)其“印跡模板”與靶點(diǎn)空間結(jié)構(gòu)的最佳適配性；黃酮類、萜類、生物堿類的MCI相似度達(dá)0.988 7、0.970 1、0.940 7，且10個(gè)對(duì)照品與中藥群體的一階矩RSD僅1.794%，驗(yàn)證其“印跡模板”特征；動(dòng)物實(shí)驗(yàn)證實(shí)，功能性“物質(zhì)單元”可顯著改善急性血瘀大鼠血液流變學(xué)指標(biāo)（低切與中切全血黏度均顯著降低，P＜0.01）及凝血功能［活化部分凝血酶原時(shí)間（activated partial thromboplastin time，APTT）顯著延長(zhǎng)，P＜0.01］。綜合評(píng)分最高的給藥性“物質(zhì)單元”來(lái)源方劑（補(bǔ)陽(yáng)還五湯組）在改善血液流變學(xué)及凝血功能的多個(gè)方面，效果均優(yōu)于評(píng)分最低的來(lái)源（白芍組），體現(xiàn)多成分協(xié)同增效特性。結(jié)論 首次構(gòu)建“個(gè)體-結(jié)構(gòu)-功能-給藥”四維解析體系，揭示黃酮類、生物堿類及萜類成分，通過(guò)超分子“印跡模板”動(dòng)態(tài)識(shí)別機(jī)制，介導(dǎo)活血化瘀效應(yīng)的核心途徑。該研究為中藥復(fù)雜體系的質(zhì)量評(píng)價(jià)與作用模式解析提供了量化工具，推動(dòng)中藥現(xiàn)代化研究從“單一成分”向“結(jié)構(gòu)簇協(xié)同”范式轉(zhuǎn)變。

Objective Based on the supramolecular “imprinting template” theory, we developed the coupling strategy of factor rotation method and molecular connectivity index (MCI), systematically analyzed the common material basis and “structure-function” correlation mechanism of traditional Chinese medicine (TCM) for promoting blood circulation and removing stasis, and provided a new paradigm for screening quality markers (Q-Marker) of TCM. Methods UPLC was used to establish the fingerprint of 13 traditional Chinese herbs [Chuanxiong (Chuanxiong Rhizoma), Danshen (Salviae Miltiorrhizae Radix et Rhizoma), Taoren (Persicae Semen), Mabiancao (Verbenae Herba), Honghua (Carthami Flos), Yanhusuo (Corydalis Rhizoma), Yuejihua (Rosae Chinensis Flos), Niuxi (Achyranthis Bidentatae Radix), Yimucao (Leonuri Herba), Sanqi (Notoginseng Radix et Rhizoma), Baishao (Paeoniae Radix Alba, PRA), Chishao (Paeoniae Radix Rubra), Danggui (Angelicae Sinensis Radix)] for activating blood circulation and removing blood stasis and three classical compounds [Buyang Huanwu Decoction (補(bǔ)陽(yáng)還五湯, BHD), Taohong Siwu Decoction (桃紅四物湯), Xuefu Zhuyu Decoction (血府逐瘀湯)], and 26 “imprinting template” component clusters were divided as structural “material units” by matching frequency method. The factor rotation method was used to reduce the dimension of the component cluster, and six functional “material units” were extracted (the cumulative variance contribution rate was 88.37%). Combined with the normalization score, the comprehensive effect of the drug “material units” was evaluated. The MCI was calculated by TCMSP database, the structural similarity of component clusters was quantified, and the contribution of core components to drug efficacy was verified by an acute blood stasis rat model. Results The “imprinting template” component cluster of traditional Chinese medicine for promoting blood circulation and removing blood stasis could be divided into six functional “material units”, of which the first three functional unit (66.85%) was dominated by flavonoids, terpenoids, and alkaloids. Due to the high abundance of flavonoids, terpenoids and alkaloids, the comprehensive score (2.44) of BHD was significantly better than that of other prescriptions, which confirmed the best fit between its “imprinting template” and the spatial structure of the target. The MCI similarity of flavonoids, terpenoids and alkaloids was 0.988 7, 0.970 1, 0.940 7, and the first moment RSD of 10 reference substances and traditional Chinese medicine group was only 1.794%, which verified their “imprinting template” characteristics. Animal experiments confirmed that functional “material unit” could significantly improve hemorheology (whole blood viscosity decreased significantly in low cut and middle cut, P < 0.01) and coagulation function (APTT prolonged significantly, P < 0.01) in rats with acute blood stasis. BHD, which ranked the highest in the normalized score of “substance unit”, improved hemorheology and coagulation function better than PRA, which reflected the synergistic effect of multi-components. Conclusion This study establishes a four-dimensional analysis system of “individual-structure-function-drug delivery” for the first time, and reveals the core mechanism by which flavonoids, terpenoids and alkaloids play the role of promoting blood circulation and removing blood stasis through dynamic recognition of the macromolecular “imprinting template”. This study provided a quantitative tool for the quality evaluation and action mode analysis of the complex system of traditional Chinese medicine, and promoted the transformation of the research on traditional Chinese medicine modernization from “single component” to “structure cluster synergy” paradigm.

國(guó)家自然科學(xué)基金項(xiàng)目（82274215）；湖南省重點(diǎn)研發(fā)計(jì)劃項(xiàng)目（2022SK2014）；湖南省自然科學(xué)基金項(xiàng)目（2022JJ30453）；湖南中醫(yī)藥大學(xué)2022年度校級(jí)“揭榜掛帥”專項(xiàng)（2022-12-27-1）；湖南省自然科學(xué)基金青年項(xiàng)目（2021JJ40398）