Study On Chemical Constituents And Biological Activity Of Shanhuoxiang

The Shanhuoxiang,also known as Xuejianchou,is a plant of the genus Labiatae in the Teucrium family.It was first recorded in the Qing Dynasty's"Medicinal herbs",which has the effect of cooling blood,stopping bleeding,detoxifying and detumescencing.Current clinical studies have reported that Shanhuoxiang has therapeutic effects on viral conjunctivitis,gynecological uterine bleeding,hypoglycemic and antibacterial effects.Based on the the records of traditional medicine and combined the comprehensive utilization of Shanhuoxiang,we explored the chemical composition and pharmacological activity of it.The in vitro hemostatic test method was used to preliminarily screen out the haemostatic site of Shanhuoxiang for the further application of hemostatic drugs.Meanwhile,the anti-tumor activity of the Shanhuoxiang is also tested,which provides a theoretical basis for further development and utilization of the Shanhuoxiang.It is based on the research of Shanhuoxaing,using ultrasonic heating of ethanol solvent extraction method,repeated use of gel column chromatography,silica gel column chromatography,thin layer chromatography and many other technical means,single 11 compounds were isolated from the three extracts of the Shanhuoxiang.The structure of 20?-hydroxy ursolic acid?1?,?–sitosterol?2?,lupeol?3?,oleanolic acid?4?,luteolin?5?,quercetin?6?,apigenin?7?,daucosterol?8?,apigenin-7-O-glucoside?9?were identified by NMR,of which compounds?1?,?3?,?7?,?9?were first isolated from the Shanhuoxiang.Compounds?10?and?11?is not identified yet.The in vitro hemostatic test method was used to preliminarily screen out the haemostatic site of Shanhuoxiang.The in vitro clotting activity of the petroleum ether,ethyl acetate,n-butanol fraction,and aqueous phase extracts of Shanhuoxiang were investigated by using the method of plasma recalcification time,test tube and coagulation plate.The results showed that the ethyl acetate extract could significantly shorten the time of coagulation and plasma calcium recovery,and the n-butanol extract also had some effect on shortening the time of coagulation and plasma calcium recovery.Compared with in vitro plasma calcium recovery time,Petroleum ether extract had no significant effect on the time of coagulation and plasma calcium recovery in vitro.The aqueous phase extract had no significant effect on the time of plasma calcium recovery,but it dcould shorten the coagulation time in vitro.The results showed that ethyl acetate and n-butanol extracts of Shanhuoxiang had significantly hemostatic effects.The petroleum ether and water phase extract was not distinct.It is necessary to further explore its hemostatic effect in vivo to further study the blood clotting effect of shanhuoxiangMTT and CCK-8 methods were used to detect the antitumor activity of the four extracts of Shanhuoxiang and four single 4 compounds.The antitumor experiment was carried out in vitro by using the cell lines of human esophageal cancer cell EC-109 and human cervical cancer cell Hela.The results showed that the n-butanol and ethyl acetate extracts of the Shanhuoxiang had significant inhibitory effects on both of the cancer cells.According to the calculation,the IC₅₀ values of ethyl acetate and n-butanol extracts to EC-109 cells were 46.69?g/mL and 30.07?g/mL respectively.The IC₅₀ values for Hela cells were 40.56?g/mL and 37.87?g/mL respectively.The anti-tumor activity of four compounds of lupeol,apigenin,apigenin-7-O-glucoside,and 20?-hydroxyursolic acid were determined.The results showed that all of the four compounds had significant antitumor activity.The IC₅₀ values of four compounds for EC-109 cells were 1.098 mg/mL,0.2705 mg/mL,0.3191 mg/mL and 2.279 mg/mL respectively.The IC₅₀ values of four compounds for Hela cells were1.4970 mg/mL?0.2325 mg/mL?0..2735 mg/mL?1.0975 mg/mL respectively.