| Rosa xanthina Lindl.,also known as yellow spines mold,Rosa flowers,is a member in the Rosa genus of Rosaceae family.It is widely distributed in Shanxi,Shaanxi,Hebei,Liaoning,Jilin and other provinces in China.The fruits of Rosa xanthina Lindl.have the function of regulating qi-flowing for strengthening spleen.Its flowers,fruits,leaves and root barks have been used to treat Stubborn gonorrhea,bacterial dysentery or women irregular menstruation.At present,it has the extensive and profound research about Rosa davurica Pall,in the same genus as Rosa xanthina Lindl.in the domestic and foreign literature.However,it is less in terms of research about Rosa xanthina Lindl.in chemical constituents,biological activity and resource development.In this paper,the present study aims to investigate chemical constituents from Rosa xanthina Lindl.and the mechanism of anticoagulant components by the prothrombin time(PT),activated partial thromboplastin(APTT)assay,thrombin time(TT),time assay of hydrolysis of fibrinogen,inhibition rates assay on thrombin and solubility assay of fibrin.The main contents are as follows:Part one:Six compounds were obtained from the 75%ethanol extract of the fruits of Rosa xanthina Lindl by liquid-liquid extraction,silica gel,sephadex LH-20,MDS and semi-preparative HPLC technology.Based on physicochemical properties and spectroscopic methods,their structures were elucidated as quercetin-3-O-α-L-rhamnopyranoside(1),quercetin-3-O-(6"-O-trans-p-coumaroyl)-β-D-glucopyranoside(2),kaempf erol-4’-O-β-D-glycopyranoside(3),quercetin-4’-O-β-D-glucopyranoside(4),1,2-bis(4-hydroxy-3-methoxyphenyl)-1,3-propanediol(5),2,3-dihydroxy-1-(3-hydroxy-4-methoxyphenyl)-1-propanone(6).Compounds of 1-5 were isolated from the plant for the first time,and compound 6 was isolated from this genus for the first time.Part two:The study is investigated their effects on anticoagulation from Quercitrin and Helichrysoside.First of all,experimental measurements of coagulation index(PT,APTT,TT)showes that Quercitrin(0.89-10.71 mM)significantly prolonged PT(P<0.01),and with the increasing concentrations,PT gradually extended for 13.3-24.ls.Helichrysoside(1.31-2.62 mM)could make PT(P<0.01)significantly prolonged 14.3-20.7s.Quercitrin(2.68-7.14 mM)could make APTT(P<0.01)significantly prolong 41.5-60.7s.APTT of Helichrysoside(0.66-3.93 mM)were 41.2-142.9 s.Different concentrations of Quercitrin(0.89-7.14 mM)and Helichrysoside(0.66-3.93 mM)could significantly reduce the thrombin time(P<0.01),respectively 18.3-14.3s and 18.0-9.4s,in a dose-dependent manner.Moreover,time assay of hydrolysis of fibrinogen showed that Quercitrin(0.89-3.57 mM)was able to significantly prolong the thrombin time(P<0.01),which was 36.2-41.2s.In addition,with the increase of concentrations(0.08-5.36 mM)of Quercitrin,the inhibition rates of thrombin increased gradually and showed dose dependence.When the concentration of Quercitrin was 1.34 mM,the inhibition rates could reach 98.6%.At last,by the fibrin plates method,experimental results indicate that different concentrations of Quercitrin(3.57,7.14 and 14.29 mM)and Helichrysoside(2.62,5.25 and 10.49 mM)have no effect on the dissolution of fibrin,compared with urokinase.The conclusions are drawn as follow:Quercitrin and Helichrysoside can significantly prolong PT,APTT and shorten TT,indicating that they have a different regulation of coagulation system.By time test of hydrolysis of fibrinogen,inhibition rates assay on thrombin,as well as solubility assay of fibrin,it is indicated that anticoagulant mechanism is that Quercitrin affect the process of transformation of fibrinogen to fibrin.In addition,effect and mechanism study of which TT is short need further illumination. |
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