Studies On The Chemical Components In Bioactive Part And Quality Control Of Fructus Livistonae Chinensis

Fructus Livistonae Chinensis is the dried ripe fruit of Livistona chinensis R.Br.It is sweet,tasteless,astringent in flavour and mild in nature.Its actions include softening and dissolving hard masses,stopping bleeding by astringency.It can be used for abdominal lumps,stagnation of qi,metrorrhagia,Traumatic injury,subhyaloid hemorrhage et al. Modern research had delicated that Fructus Livistonae Chinensis possessed the effect of anti-tumor,and widely used for the treatment of cancer in the folk,such as esophagus cancer,nasopharyngeal cancer,chorioadenoma,leukaemia,choriocarcinoma,lung cancer. But researches on the effective part about chemical constituents and pharmacology haven't been reported.In order to obtain a more comprehensive understanding of its effective physiological constituents,reveal the substantial foundation of anti-tumor effect and provide scientific basis of producing medicine reasonably,a phytochemical study has been carried out.The fore-experiments of its chemical components showed that it contained phenols, anthraquinones,coumarin,steroids,triterpenoids,saponin,amino acids,et al.The 70%alcoholic extracts obtained from the dried fruits were divided into four portions including petroleum ether extracts,ethyl acetate extracts,n-Butyl alcohel extracts and water extracts for the first time.MTT assay and ELISA were used to detect the proliferation and the secretion of vascular endothelial growth factor(VEGF) of chronic myelogenous leukemia K562,ovarian neoplasm SKOV3,colon carcinoma HT-29 and bladder cancer T24 cell lines respectively.The expression of fetal liver kinase(Flk-1) protein and mRNA in endothelial cells induced by VEGF was analyzed by Western blotting and RT-PCR.The results showed that the ethyl acetate part could selectively inhibit the proliferation of HT-29 and T24 cell lines in a dose-dependent manner(the maximum inhibitory rate were 74.66%and 86.52%),and could significantly inhibit the secretion of VEGF protein of the four tumor cell lines.The petroleum ether part had similar inhibitory effect as well as the ethyl acetate part,but its potency is inferior to the ethyl acetate part(the maximum inhibitory rate were 43.80%and 38.67%),The water-soluble and n-butanol parts had no effect on different tumor cell lines.The ethyl acetate part could inhibit significantly the expression of Flk-1 protein and mRNA of HUVEC induced by VEGF.The ethyl acetate part has fine anti-tumor activity,and its mechanism might be associated with reducing VEGF protein secretion and inhibiting the expression of Flk-1 mRNA and protein.Effective parts were seperated and purified by chromatographic methods. Compounds were identified by determining melting point,UV,IR,MS,¹H-NMR, ¹³C-NMR,et al.10 compounds had been isolated from ethyl acetate extract of Fructus Livistonae Chinensis,and 5 compounds were identified:(-) -epiafzelechin(â… ), daucosterol(â…¡),β-sitosterol(â…¢),emodin(â…¥),tricin(â…¦).The compoundâ…¥were obtained from the plant for the first time.For the first time,we analyzed the chemical constituents of petroleum ether extracts of Fructus Livistonae Chinensis and the constituents were separated and identified by GC-MS.The results showed that 11 compounds were identified. Unsaturated fatty acids constituted 52.28%of the total fatty acids,which includeω-3 Polyunsaturated fatty acid:α-linolenic acid,EPA.For the first time,the content of tricin in Fructus Livistonae Chinensis was determined by HPLC.The results showed the HPLC method was sentives and specifive. Finger printing research on Fructus Livistonae Chinensis was conducted.The results showed that 31 peaks could denote the fundamental substance of Fructus Livistonae Chinensis,the research method was advanced and feasible,which indicated the finger printing is somehow meanmingful as a measure of quality control when the chemical components are unknown.The research provided the scientific fundation to reveal the substantial foundation of anti-tumor effect,establish scientific evaluating method of quality control,achieve medicine material continuously reasonable exploitation,exploit antitumor new medicine of Chinese herbal medicine.