[Comparative analysis of anti-thrombotic effect and metabonomic mechanism of aqueous extract of leech before and...

This study compared the pharmacodynamics and metabolomics of aqueous extracts of leech before and after talc processing, focusing on their anti-thrombotic effects in thrombotic rats, intending to clarify the impact of talc processing on the efficacy and mechanism of leech. A carrageenan-induced rat tail thrombosis model was employed. Drugs were administered for five days before modeling, and 24 hours after modeling, the black tail length/rate was measured. Blood samples were collected to assess coagulation parameters, platelet aggregation rates(maximum aggregation rate, MAR; average aggregation rate, AAR), pathological sections, and blood biomarkers(adenylate cyclase, AC; cyclic adenosine monophosphate, cAMP). Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used in combination with orthogonal partial least squares-discriminant analysis(OPLS-DA) to screen differential metabolites. Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis was applied to elucidate metabolic regulatory mechanisms. The results show that compared with the blank group, the model group exhibited significantly increased black tail length and rate, significantly shortened coagulation time, markedly increased fibrinogen(FIB) content, significantly increased MAR and AAR, and decreased expression of AC and cAMP. The pathological section examination shows that thrombi occupied the vessels. Compared with the model group, raw and talc-processed leech extracts-treated groups show significantly decreased black tail length and rate, significantly prolonged coagulation time, markedly decreased FIB content, significantly reduced MAR and AAR, and increased AC and cAMP levels. The pathological section examination shows that the thrombus in the tail section of the rats in the raw and talc-processed leech extract groups was significantly reduced. Compared with the blank group, 57 differential metabolites were screened out in the model group, among which 40 and 39 biomarkers were modulated by the raw and talc-processed leech. The raw leech mainly affects glycerophospholipid metabolism, arachidonic acid metabolism, and glycerolipid metabolism. The metabolism of glycerophospholipid, the synthesis of phenylalanine, tyrosine, and tryptophan, arachidonic acid metabolism, phenylalanine metabolism, and glycerolipid metabolism are mainly affected by talc-processed leech. In conclusion, both raw leech and talc-processed leech have good antithrombotic effects, with talc-processed leech having a stronger effect. The two leech products can significantly reverse the metabolic abnormalities in rats with tail thrombosis, and the mechanism may be related to the regulation of glycerophospholipid metabolism, arachidonic acid metabolism, and glycerolipid metabolism.