Table_9.DOCX

MaZiRenWan (MZRW, also known as Hemp Seed Pill) is a Chinese Herbal Medicine which has been demonstrated to safely and effectively alleviate functional constipation (FC) in a randomized, placebo-controlled clinical study with 120 subjects. However, the underlying pharmacological actions of MZRW for FC, are still largely unknown. We systematically analyzed the bioactive compounds of MZRW and mechanism-of-action biological targets through a novel approach called “focused network pharmacology.” Among the 97 compounds identified by UPLC-QTOF-MS/MS in MZRW extract, 34 were found in rat plasma, while 10 were found in rat feces. Hierarchical clustering analysis suggest that these compounds can be classified into component groups, in which compounds are highly similar to each other and most of them are from the same herb. Emodin, amygdalin, albiflorin, honokiol, and naringin were selected as representative compounds of corresponding component groups. All of them were shown to induce spontaneous contractions of rat colonic smooth muscle in vitro. Network analysis revealed that biological targets in acetylcholine-, estrogen-, prostaglandin-, cannabinoid-, and purine signaling pathways are able to explain the prokinetic effects of representative compounds and corresponding component groups. In conclusion, MZRW active components enhance colonic motility, possibly by acting on multiple targets and pathways.麻子仁丸（MaZiRenWan，简称MZRW，又称Hemp Seed Pill）是一种中草药，已在一项纳入120名受试者的随机安慰剂对照临床试验中被证实可安全且有效地缓解功能性便秘（functional constipation，FC）。然而，麻子仁丸治疗功能性便秘的潜在药理学作用机制仍大多未明确。本研究通过一种名为"聚焦网络药理学（focused network pharmacology）"的全新方法，系统分析了麻子仁丸的生物活性成分及其参与药效作用机制的生物学靶点。在通过超高效液相色谱-四极杆飞行时间串联质谱（UPLC-QTOF-MS/MS）从麻子仁丸提取物中鉴定出的97种化合物中，有34种可在大鼠血浆中检出，另有10种可在大鼠粪便中检出。层级聚类分析（hierarchical clustering analysis）结果显示，这些化合物可被划分为多个成分组，组内化合物结构高度相似，且大部分均来源于同一味中药材。研究选取大黄素（Emodin）、苦杏仁苷（Amygdalin）、白芍苷（Albiflorin）、厚朴酚（Honokiol）及柚皮苷（Naringin）作为各对应成分组的代表性化合物。体外实验结果表明，上述所有代表性化合物均可诱导大鼠结肠平滑肌产生自发性收缩。网络分析结果显示，乙酰胆碱（acetylcholine）通路、雌激素（estrogen）通路、前列腺素（prostaglandin）通路、大麻素（cannabinoid）通路及嘌呤（purine）信号通路中的生物学靶点，可解释代表性化合物及对应成分组的促结肠动力作用。综上，麻子仁丸的活性成分可能通过作用于多靶点及多通路，增强结肠动力。