Antithrombosis Effect On Pulmonary Thromboembolism And Blood Stasis Animal Model Of AEE And Its Anti-platelet Aggregation Mechanism

With the development of economy and society,life spans of companion animals such as dog and cat have observably been lengthened,and age-related thrombosis has been a dangerous“killer”of geriatric companion animal.However,there is not an animal-specific anti-thrombosis drug until now.Aspirin eugenol ester?AEE?is a novel synthesized drug for prevention and treatment of inflammation,headache and fever.Previous researches indicated that AEE had antithrombosis effect in carrageen-induced thrombosis model.The study is to be further confirmed by determining the protective effect of AEE on mice with pulmonary thromboembolism and the antithrombosis effects of AEE in blood stasis rat model.Based on thrombosis formation mainly caused from abnormal platelet aggregation,anti-platelet aggregation mechanisms of AEE were investigated.1.Mice model with pulmonary thromboembolism was successfully established by using collagen and epinephrine.Lung HE sections indicated that there were some pathological changes including thrombosis in blood vessels.AEE could increase the survival rates and decrease the lung/body coefficient on mice with pulmonary thromboembolism,which indicated that AEE possessed some protective effect on mice with pulmonary thromboembolism.2.Rat blood stasis model was established by using epinephrine and ice water.AEE could significantly increase PT,APTT,TT,FIB,PLT,PCT and 6-keto-PGF_1?,and decrease whole blood viscosity,plasma viscosity and TXB₂.These results showed that AEE possessed good antithrombosis effect in rat blood stasis model.3.The anti-platelet aggregation mechanisms of AEE were assayed in vitro by platelet agonist inducing platelet aggregation from healthy rat blood.AEE could not affect LDH content,which indicated that the anti-platelet effect of AEE was not associated with its cytotoxicity.AEE markedly attenuated granule secretion markers?P-selectin expression and ATP release?,[Ca²⁺]_i mobilization and TXB₂ formation.AEE also attenuated CD40L activation,suppressed akt,JNK1 and ERK2phosphorylation,and recovered Sirt1 expression,but did not affect p38,VASP^Ser157 and VASP^Ser239activation and cAMP level.These results indicated that AEE inhibits agonist-induced platelet aggregation via PI3K/Akt,Sirt1/CD40L and MAPK pathways,but not cAMP-mediated VASP phosphorylation.In addition,AEE could not affect bleeding time in mice,which inhibits platelet aggregation without bleeding risk.In conclusion,AEE has significant antithrombosis activity,which depended on the suppression of AEE on platelet aggregation via PI3K/Akt,MAPK and Sirt 1/CD40L pathways.