Fu Jinzhou, Zhang Guoli, Peng Hongbing, Xu Zijie, Liu Yimei, Chen Keli, Li Juan
Hubei Key Laboratory of Resources and Chemistry of Chinese Medicine, School of Pharmacy, Hubei University of Chinese Medicine, Wuhan, China.
Hubei Shizhen Laboratory, Wuhan, China.
PLoS One. 2025 May 14;20(5):e0320165. doi: 10.1371/journal.pone.0320165. eCollection 2025.
Aspongopus chinensis Dallas holds both medicinal and culinary significance in China. Notably, in regions such as Guizhou and Yunnan, it has been traditionally used as an ethnic remedy for treating various conditions, including stomach coldness, pain, kidney deficiencies, and impotence, among other ailments. This study aims to explore the chemical constituents and anticoagulant activity of A. chinensis.
Ultraperformance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was employed for the UPLC fingerprint analysis. The isolated compounds underwent in vitro assays to their evaluate anticoagulant properties and effects on protein fibril activity. Network pharmacology was utilized to predict potential anticoagulant targets. Furthermore, animal experiments were conducted to measur coagulation factors and assess the in vivo anticoagulant activity.
Our findings indicate that 10 compounds were identified through UPLC-Q-TOF-MS analysis, with four compounds (uracil; 6-hydroxyquinolinic acid; 1,4-dihydro-4-oxoquinoline-2-carboxylic acid; delicatuline B) were isolated and identified from the n-butanol extract of A. chinensis. Furthermore, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (PLS-DA) of the UPLC fingerprint data revealed significant differences between A. chinensis and its similar insects. Notably, 1,4-dihydro-4-oxoquinoline-2-carboxylic acid has been identified as a potential standard reference substance for content determination. The isolated compounds showed anticoagulant properties, and the mRNA expression levels of MMP9 and PTGS2 were significantly reduced in LPS-induced RAW264.7 cells, further supporting the network pharmacology analysis. Animal experiments confirmed the potent anticoagulant effects of A. chinensis, likely associated with the intrinsic coagulation pathway and the inhibition of platelet aggregation.
This research provides a quality assessment method for A. chinensis, and has also demonstrated its anticoagulant functions and substance basis.
中华稻缘蝽在中国兼具药用和食用价值。值得注意的是，在贵州和云南等地，它传统上被用作民族药物来治疗多种病症，包括胃寒、疼痛、肾虚和阳痿等其他疾病。本研究旨在探索中华稻缘蝽的化学成分和抗凝活性。
采用超高效液相色谱-串联四极杆飞行时间质谱（UPLC-Q-TOF-MS）进行UPLC指纹图谱分析。对分离得到的化合物进行体外试验，以评估其抗凝特性和对蛋白质纤维活性的影响。利用网络药理学预测潜在的抗凝靶点。此外，进行动物实验以测量凝血因子并评估体内抗凝活性。
我们的研究结果表明，通过UPLC-Q-TOF-MS分析鉴定出10种化合物，其中从中华稻缘蝽的正丁醇提取物中分离并鉴定出4种化合物（尿嘧啶；6-羟基喹啉酸；1,4-二氢-4-氧代喹啉-2-羧酸；精致素B）。此外，UPLC指纹图谱数据的主成分分析（PCA）和正交偏最小二乘法判别分析（PLS-DA）显示中华稻缘蝽与其相似昆虫之间存在显著差异。值得注意的是，1,4-二氢-4-氧代喹啉-2-羧酸已被确定为含量测定的潜在标准参考物质。分离得到的化合物具有抗凝特性，并且在脂多糖诱导的RAW264.7细胞中，MMP9和PTGS2的mRNA表达水平显著降低，进一步支持了网络药理学分析。动物实验证实了中华稻缘蝽具有强大的抗凝作用，这可能与内源性凝血途径和血小板聚集的抑制有关。
本研究为中华稻缘蝽提供了一种质量评估方法，同时也证明了其抗凝功能和物质基础。
