| Purpose:Emblica Euphorbiaceae Phyllanthus emblica L. is the Tibetan conventional medicine. Phyllanthus emblica L. has many pharmacological effects, and more and more scholars from various countries have extensive attention of the anti-cancer effect. In this paper, for the study of Chinese medicine Phyllanthus emblica L., we use modern analytical techniques, and in vitro and in vivo efficacy of immunization experiments, to selecte anti-tumor activity component of Tibetan medicine Phyllanthus emblica L., and study its material basis, anti- tumor mechanism of the initial chemical composition of Phyllanthus emblica L. and the ownership of the main peak of different areas of P. emblica and the extract of total phenolic acids and the flavonoids,in order to provide scientific basis for the development of new anticancer drugsMethods:Using serum pharmacology and pharmacology of plasma selects different ethanol extracts of P. emblica preliminarily. After administration of water extract of P. emblica,70% ethanol extract of P. emblica, and 90% ethanol extract of P. emblica, the seruma and plasma of Kunming mice were obtained. Using MTT method to detect the inhibition rates of different cancer cell lines, which is the drug-containing extracts from Phyllanthus emblica L. containing plasma and serum on, and which pharmacology of plasma, serum pharmacology experimental results were compared.By UV, determining the content of the total phenolic acids and total flavonoids of Phyllanthus emblica L. With the use of HPLC, determining the gallic acid content of the in different places. Purifying the emblica total phenolic acids and flavonoids by macroporous resin method.With the Use of MTT assay in vitro cell, determine P. emblica total phenolic acids and flavonoids on liver-cancer cells and cervical cancer cell line. To conduct a preliminary study of its in vitro pharmacodynamicsBy geometric method with the baseline design fundamentals and MTT method we determined the compatibility of anti-tumor components of he emblica total phenolic acids and flavonoids. with the baseline geometric design principle of law, after the administration of the different proportions of the total phenolic acids and flavonoids, the the mice serum were obtained. With serum IL-2 ELISA kit and serum IFN-?ELISA kit, determining the inhibition of tumor cells of P. emblica and immune regulatory role. By the combination of cell inhibition and immune regulation, analying the results of the compatibility and getting Compatibilities formulations of total flavonoids and total phenolic.The fingerprint of HPLC method using gradient elution of purified extract of P. emblica the total acid and lavonoids study is designed to compare the difference among different places, establish extracts emblica HPLC fingerprints of the representative of gallic acid and rutin, and provide a theoretical basis of internal quality control.By flow cytometry PI single labeled to detect cell cycle. Drugs can prevent the uncontrolled proliferation of cells, and block the cell in the G1 phase (ie, pre-D NA synthesis) or S phase (DNA synthesis phase), or G2 phase (DNA synthesis later.) In this experiment, by flow cytometry, total phenolic acids and flavonoids are detected in cells in which the time to stop.Result:1. Through the use of aqueous extract of Phyllanthus emblica L., Phyllanthus emblica L 70% ethanol extract, emblica extract 95% ethanol were taken after administration to mice effects on different cells in serum plasma, measured OD values show inhibition. The results showed that 70% of P. emblica plasma ethanol extract inhibited the overall higher rate of cancer cells.2. Experimental results showed that 70% ethanol emblica extracts drug-containing plasma, and inhibition rate to cancer cells of liver cancer and cervical cancer is better. In this experiment, the final selection of anti-liver cancer and cervical cancer emblica main research objective.3. By ultraviolet spectrophotometry, total flavonoids and total phenolic content of aqueous extract of Phyllanthus emblica L., Phyllanthus emblica L.70% ethanol extract of P. emblica 95% ethanol extract was determined. Total phenolic acid contents were 266.00mg /g,247.20mg/g,86.40mg/g. Total flavonoids were 56.58mg/g,59.52mg/g,54.30mg /g. Through comprehensive comparison, total phenolic and total phenolic concentrations of 70% ethanol extract are relatively high.4. By HPLC gallic acid content of different places were determined, and the experimental results show that:gallic acid content of Guizhou origin is the highest percentage per gram of raw medicinal herbs in the content of 1.578%5. Use macroporous resin to purify total phenolic acids to establish the process route, and make sure that the selection is polyamide resin, elution agent is the water column elution volume was 6 times to the volume of resin, the high ratio of column diameter is 1:5, and flow rate is 1.5mL/min. 6. Use macroporous resin purify total flavonoids and establish routes for the purification process through polyamide resin, eluent 50% ethanol,7 times the column volume of elution volume, diameter of the high ratio of resin 1:5, flow rate of 1mL/min.7. The total phenolics of purified Emblica effects on cancer cell SMMC-7721 and Hela and then the the aging curve is printed. The maximum inhibition rate is 83.9% and 92.9%.8. The total flavonoids of purified Emblica effects on cancer cell SMMC-7721 and Hela and then the the aging curve is printed. The maximum inhibition rate is 66.1% and 88.5%.9. Combination on vitro experiments, when the ratio of total phenolics and flavonoids is 34.56?g:34.78?g,the anti-cancer agents is best.We have vivo animal immunization studied, and the antioxidant properties of total phenolics instructed, for supporting the interpretation of the results of compatibility to further determination of the compatibility proportion. The results show that the total phenolic and flavonoids in emblica can improve immunity.10. By HPLC the total flavonoids and the total phenolics of the fingerprint of the emblica from different places are determinated. The results demonstrate that the quality of different origin emblica is uniform and stable.11. By using flow cytometry the cell cycle of apoptosis is determinated. Emblica can block the S phase of cancer cell. Emblica can also affect normal process of cell cycle and cell apoptosis. |
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