背景:葛根(威尔德。)Ohwi和葛根。thomsonii(Benth.)Maesen是营养药食品同源植物,广泛用于食品和保健品行业,是开发新型保健食品的优良天然材料,国内外市场潜力巨大。临床上,P.lobata和P.thomsonii用于治疗冠心病,动脉粥样硬化,脑梗塞和其他心血管疾病,抗血栓作用可能是其治疗血栓性疾病的核心作用。然而,P.lobata和P.thomsonii抗血栓特性的潜在机制尚未阐明。
方法:首先,通过高效液相色谱(HPLC)鉴定了P.lobata和P.thomsonii。建立动静脉旁路血栓形成大鼠模型。血栓干-湿重,血小板蓄积率和四个凝血指标,包括活化部分凝血活酶时间(APTT),凝血酶原时间(PT),凝血酶时间(TT)和纤维蛋白原(FIB),在血浆中检测到P.lobata和P.thomsonii抗血栓形成功能。采用网络药理学和分子对接方法获得关键靶点并验证可靠性。David6.8用于GO和KEGG分析,以探索P.lobata和P.thomsonii抗血栓形成功能的途径和潜在靶标。前列腺素I2(PGI2),血栓烷A2(TXA2),环氧合酶2(COX-2),采用酶联免疫吸附试验(ELISA)检测髓过氧化物酶(MPO)和内皮型一氧化氮合酶(eNOS)。
结果:结果表明,P.lobata和P.thomsonii可以减少大鼠的血栓干-湿重和血小板积聚,并抑制TT,APTT,FIB,和PT。一种全面的网络药理学方法成功地鉴定了P.lobata和P.thomsonii中的9种活性成分。主要活性成分包括多酚,氨基酸和类黄酮。共获得15个抗血栓功效目标,包括3个关键目标(PTGS2、NOS3、MPO)。路径分析显示10条显著相关的通路和29个生物学过程。通过上调PGI2和下调TXA2来抑制血小板聚集,抑制PTGS2以减轻炎症,并增加eNOS水平以促进血管舒张。此外,P.lobata和P.thomsonii通过增加SOD水平和显着降低MDA含量来减轻氧化应激。
结论:研究结果进一步阐明了P.lob和P.thomsonii的抗血栓作用机制,为血栓性疾病新药的开发提供了科学依据,为洛氏疟原虫和托氏疟原虫草药资源和洛氏疟原虫保健品的开发奠定了基础。
BACKGROUND: Pueraria lobata (Willd.) Ohwi and Pueraria lobata var. thomsonii (Benth.) Maesen are nutritious medicine food homology plants that are widely used in the food and health products industry and are excellent natural materials for the development of new health foods, with great potential for domestic and foreign markets. Clinically, P. lobata and P. thomsonii are used to treat coronary heart disease, atherosclerosis, cerebral infarction and other cardiovascular diseases, and antithrombotic actions may be their core effect in the treatment of thrombotic diseases. However, the underlying mechanisms of the antithrombotic properties of P. lobata and P. thomsonii have not been clarified.
METHODS: First, P. lobata and P. thomsonii were identified by high-performance liquid chromatography (HPLC). An arteriovenous bypass thrombosis rat model was established. Thrombus dry‒wet weight, platelet accumulation rate and the four coagulation indices, including activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT) and fibrinogen (FIB), were detected in plasma to manifest the P. lobata and P. thomsonii antithrombotic function. Network pharmacology and molecular docking methods were used to obtain key targets and verify reliability. David 6.8 was used for GO and KEGG analyses to explore pathways and potential targets for P. lobata and P. thomsonii antithrombotic functions. Prostaglandin I2 (PGI2), thromboxane A2 (TXA2), cyclooxygenase 2 (COX-2), myeloperoxidase (MPO) and endothelial nitric oxide synthase (eNOS) were tested by enzyme-linked immunosorbent assay (ELISA).
RESULTS: The results indicated that P. lobata and P. thomsonii can reduce thrombus dry‒wet weight and platelet accumulation in rats and inhibit TT, APTT, FIB, and PT. A comprehensive network pharmacology approach successfully identified 9 active ingredients in P. lobata and P. thomsonii. The main active ingredients include polyphenols, amino acids and flavonoids. A total of 15 antithrombotic function targets were obtained, including 3 key targets (PTGS2, NOS3, MPO). Pathway analysis showed 10 significant related pathways and 29 biological processes. P. lobata and P. thomsonii inhibited platelet aggregation by upregulating PGI2 and downregulating TXA2, inhibited PTGS2 to reduce inflammation, and increased the level of eNOS to promote vasodilation. In addition, P. lobata and P. thomsonii alleviated oxidative stress by increasing SOD levels and significantly decreasing MDA contents.
CONCLUSIONS: The results of the study further clarify the antithrombotic mechanism of action of P. lobata and P. thomsonii, which provides a scientific basis for the development of new drugs for thrombogenic diseases and lays the foundation for the development of P. lobata and P. thomsonii herbal resources and P. lobata and P. thomsonii health products.