Study On Hemostatic Activity Constituents And Mechanisms Of Toddalia Asiatica Root Bark

Objective:To study the chemical constituents of n-butanol part of the root bark of Toddalia asiatica in the family Rutaceae,and to perform a series of research on Toddalia asiatica Root Barkaround the coagulation system,fibrinolysis system and platelet system,aiming to more scientifically explain its material basis and hemostatic mechanisms.Methods:1.The n-butanol parts of the toddalia asiatica were isolated and purified by macroporous resin,C18,silica gel and Sephadex LH-20 column chromatography.Their structures were elucidated on the basis of physicochemical properties,NMR,MS and literature comparisons.2.The hemostatic mechanism of toddalia asiatica was studied by measuring the parameters of blood agglutination,fibrinolytic system and CF-PK-PD analysis.Results:1.Eighteen compunds were obtained from n-butanol part of toddalia asiatica root bark.Fourteen compounds of them were identified:pimpinellin(1),Bergapte(2),alloimperatorin(3),isopimpinellin(4),mono(2-ethylhexyl)terephthalate(5),(+)-toddalolactone(6),nitidine(7),phellopterin(8),ferulic acid methyl ester(9),4-methoxycinnamic acid(10),hesperidin(11),methyl trans-4-Hydroxycinnamate(12),moellendorffiline(13),avenalumic acid methyl ester(14),Compounds 3,9,12,and 14 were isolated from this genus for the first time.Compounds 5,10,and 13 were isolated from this family for the first time.2.It was found that the hemostatic activity of toddalia asiatica root bark was achieved by activating the endogenous and exogenous coagulation pathways and increasing the sensitivity of the fibrinolytic system.By CF-PK-PD analysis,six major natural products were found in the plasma of hemorrhagic rat models.They were:Skimmianine,γ-fagarine,lomatin,pimpinellin,5-methoxy-8-hydroxy-psoralen.isopimpinellin,The compounds lomatin and methoxy-8-hydroxy psoralen had significant effects on the regulation of endogenous coagulation pathway and fibrinolytic system.Isopimpinellin and pimpinellin have a good PK-PD association with FIB in the coagulation system.At the same time,the calculation and prediction of isopimpinellin were Emax4.87mg/m L,EC501.39μg/m L,Ke0 0.811/h;forpimpin-ellin,Emax 4.29mg/m L,EC50 2.47μg/m L,and Ke0 0.771/h.After investigation on the indexes of fibrinolysis system,the levels of DD and FDP in the model group were found significantly lower than those in the blank control group(P < 0.05).After treatment with EA and n-B,the levels of DD and FDP were significantly increased(P < 0.05).The effect of the n-B extract on the level of FDP was prominent,while the regulation of DD levels in EA extractwas significantly biased.Examination of the platelet system under effect of the compounds revealed that compounds 1 and 11 had significant antiplatelet aggregation activity.Conclusion:1.This study has further enriched and perfected the chemical constituents of toddalia asiatica root bark.2.The toddalia asiatica is to achieve hemostasis and coagulation mainly through coagulation system and fibrinolysis system,the hemostatic and coagulant effects of coagulation system were related to the active constituents of the compounds lomatin,isopimpinellin,pimpinellin and 5-methoxy-8-hydroxy psoralen.