Urinary metabolomics study the mechanism of Taohong Siwu Decoction intervention in acute blood stasis model rats based on liquid chromatography coupled to quadrupole time-of-flight mass spectrometry

Taohong Siwu Decoction (TSD) is a classic prescription in traditional Chinese medicine and is widely used to promote blood circulation to remove blood stasis. However, the effect mechanisms are not yet well understood. Here, a urinary metabolomic approach based on liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC/Q-TOF-MS) was conducted to explore the changes in the endogenous metabolites and to assess the integral efficacy of TSD on acute blood stasis model rats. Then, parameters for hemorheology and coagulation functions were detected. Principal component analysis (PCA) and orthogonal partial least squares discriminate analysis (OPLS-DA) was used to investigate the global metabolite alterations and to evaluate the preventive effects of TSD in rats. Potential metabolite markers were found using OPLS-DA and t-test. Furthermore, metabolic pathway analysis was performed to construct metabolic networks. The results showed that TSD could significantly decrease whole blood viscosity and plasma viscosity. It also significantly prolonged partial thromboplastin time (APPT) and prothrombin time (PT), increased thrombin time (TT) and lowered fibrinogen content (FIB). Moreover, 24 potential metabolite markers of acute blood stasis were screened, and the levels were all reversed to different degrees after TSD administration. In metabolic networks, amino acid metabolism (arginine and proline metabolism; histidine metabolism; alanine, aspartate, and glutamate metabolism; phenylalanine, tyrosine, and tryptophan biosynthesis; phenylalanine metabolism) and lipid metabolism (glycerophospholipid metabolism; linoleic acid metabolism; alpha-linolenic acid metabolism) were closely related with the intervention mechanism of TSD on acute blood stasis. The urinary metabolomic approach can be applied to clarify the mechanism of TSD in promoting blood circulation to remove acute blood stasis and to provide the theoretical basis for further research on the therapeutic mechanism of TSD in clinical practice.

桃红四物汤（TSD）是中医经典方剂，广泛用于活血化瘀。然而，其作用机制尚未完全明确。在此，采用基于液相色谱 - 四极杆飞行时间质谱（LC/Q - TOF - MS）的尿液代谢组学方法，探究内源性代谢物的变化，并评估TSD对急性血瘀模型大鼠的整体疗效。然后，检测血液流变学和凝血功能相关参数。运用主成分分析（PCA）和正交偏最小二乘法判别分析（OPLS - DA）研究整体代谢物的变化，并评估TSD对大鼠的预防作用。通过OPLS - DA和t检验发现潜在的代谢物标志物。此外，进行代谢通路分析以构建代谢网络。结果表明，TSD可显著降低全血粘度和血浆粘度，还可显著延长部分凝血活酶时间（APPT）和凝血酶原时间（PT），增加凝血酶时间（TT）并降低纤维蛋白原含量（FIB）。此外，筛选出24种急性血瘀的潜在代谢物标志物，TSD给药后其水平均有不同程度的逆转。在代谢网络中，氨基酸代谢（精氨酸和脯氨酸代谢；组氨酸代谢；丙氨酸、天冬氨酸和谷氨酸代谢；苯丙氨酸、酪氨酸和色氨酸生物合成；苯丙氨酸代谢）和脂质代谢（甘油磷脂代谢；亚油酸代谢；α - 亚麻酸代谢）与TSD对急性血瘀的干预机制密切相关。尿液代谢组学方法可用于阐明TSD活血化瘀治疗急性血瘀的机制，为进一步研究TSD在临床实践中的治疗机制提供理论依据。