Studies On The Active Constituents And The Quality Control Of Livistona Chinensis

Cancer is a severe health and safety menace to human being, and the current morbidity and mortality showed an tendency to increase year by year. Therefore it is of great significance to study antitumor drugs which are high efficiency, low toxicity. And presently, natural antitumor drug is one of focal topics in the research and development of the anticancer drugs.Chinese fanpalm seed is the seed of Livistona chinensis (Jacq.) R. Br. which is palm plants. It is not only widely used in treating various cancers in Fujian folk, and modern pharmacological studies also confirmed its significant effect for anticancer. However, less research was carried out on the seed of Livistona chinensis (abbreviated as L. chinensis ), in addition to a few study on chemical constituents or the anticancer activity of crude extracts. Therefore, the relevant pharmaceutical research on L. chinensis is necessary to be furthered, which will provide the foundation for its anticancer medicinal research. So this experiment was beginned with the pharmacognosy study of L. chinensis, then the anticancer activities of the different parts or components from L. chinensis. Established HPLC-DPPH method to screen the effective antioxidative peaks of the extract from L. chinensis, and identify the corresponding structures by liquid chromatography-mass spectrometry. Developed the further research on chemical constituents of L. chinensis, and studied the antitumor activity of the obtained constituents in vitro screening and in vivo animal experiment by transplanted tumors. The pharmacokinetics of the main active constituent was investigated in vivo. Quality assessment was established by the content determination and fingerprint of L. chinensis. The main research content and academic contributions were as follows:1. The study was carried out on the textual research of L. chinensis, original plant morphology, distribution, morphological identification, physicochemical identification, microscopic characteristics, and content determination. The study provides pharmacognosy reference of L. chinensis for medicinal use.2. After preliminary extraction and separation, four different polar fractions were obtained. Screening antitumor activities of various fractions in vitro, ethyl acetate fraction and butanol fraction was showed better activity. Then using transplanted tumors in mice to confirm antitumor effects in vivo.3. Established HPLC-DPPH method to screen the antioxidative constituents of the natural products effectively. And the method was used to screen the antioxidant constituents of L. chinensis for the first time, and the result showed that there are seven main constituents exhibited good antioxidant activity. The study revealed the antioxidant activity of L. chinensis, and demonstrate the major antioxidant pesks of L. chinensis for the first time. It also provided a feasible means of screening for the antioxidant components from natural products.4. In view of the result of screening for the antioxidant constituents by HPLC-DPPH, six major antioxidant constituents were identified from the total extract of L. chinensis by liquid chromatography-mass spectrometry (LC/MS) according to the mass featured fragments and fragment pathway, and five compounds mentioned above were detected from this plant for the first time.5. Phytochemistry separation was carried out, twenty five compounds were isolated from L. chinensis. The structure of eighteen compounds were identified by means of 1~H-NMR, (13)~C-NMR, Mass and IR, etc. Eight compounds mentioned above were isolated from this plant for the first time, including three compounds were isolated from this famly for the first time.6. After screenning the antitumor activity of the isolated components and constituents in vitro, tricin showed better activity. Apoptotic effect of tricin was detected with AO/EB fluorescent staining, DNA fragmentation by agarose gel electrophoresis analysis, CDX2 expression by immune histochemistry method, the distribution of cell cycle by flow cytometric assay, the expression of Bcl-2 and Bax by Western Blot. Then the antitumor activity of tricin was confirmed in vivo. This study reveals tricin may be one of the major anticancer constituents.7. An HPLC method for determination of tricin concentration in rats'plasma was established, and pharmacokinetics of tricin in rats after oral administration was firstly studied. This study provided a useful reference on the process in vivo for tricin. 8. An HPLC-UV method for the simultaneous determination of six major constituents isolated from L. chinensis was establish, the contents of these constituents in L. chinensis from ten different habitats have been measured.This method can be used as quality control for quantitative analysis of L. chinensis.9. To established fingerprint analysis of L. chinensis by HPLC-UV, the precision, repeatability and stabilization of this method were investigated. The consistency of the fingerprint of ten batches of L. chinensis from different habitats were studied. This study will be provided as a more comprehensive qualitative evaluation for quality control and distignishing the L. chinensis.