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Polygonum orientale treatment of liver tumors chemomics and Mechanism

Author: ChouJing
Tutor: MengXianSheng
School: Liaoning University of Traditional Chinese Medicine
Course: Pharmacognosy
Keywords: Polygonum orientale Constituent Against liver cancer Pharmacodynamic
CLC: R285
Type: Master's thesis
Year: 2011
Downloads: 62
Quote: 0
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Abstract


Objective: Polygonum orientale treatment of liver cancer drug research for the national project \object of study, the use of modern separation and analytical techniques, the use of the active components screening methods to extract Polygonum orientale anti-hepatoma chemicals group, separation and purification; combined in vitro and in vivo pharmacodynamic study efficacy, screening Polygonum orientale anticancer effective components; preliminary study of its mechanism of action. An effective anti-cancer component of Polygonum orientale more in-depth research, theoretical and experimental basis for the development of traditional Chinese medicine Polygonum orientale, lay the material component compatibility of Chinese medicine treatment of liver cancer drug research and experimental basis. MATERIALS AND METHODS: (1) using high performance liquid chromatography, Polygonum orientale known anti-cancer active ingredients taxifolin content of quercetin and a cream rate and in vitro efficacy as index by single factor -trips and the L 9 (3 4 ) orthogonal experiment, the extraction process of Polygonum orientale preferred. HPLC was used to filter out static experiments purified active ingredient of Polygonum orientale macroporous resin, dynamic experiments to determine the best separation and purification conditions macroporous resin component of effective substances. 3 MTT assay, as the research object to of Polygonum orientale effective chemicals group, select human hepatoma SMMC-7721 and HepG2 cells were observed of Polygonum orientale effective chemicals group on the proliferation. Using serum pharmacological method detection Polygonum orientale chemicals group containing serum on proliferation of hepatoma cells suppression. MTT assay, Polygonum orientale anti-hepatoma groups of substances selected for the study, human liver cells L02, observe the effects of different time relative growth degree, determine the hepatotoxicity. 5. Of Polygonum orientale effective chemicals group acute toxicity test the measured mouse maximum tolerated dose (MTD). H22 mice transplanted tumor models with different doses of Polygonum orientale chemicals group were gavage treatment, to observe the impact on the growth of liver tumors. High performance liquid wavelength integration chromatography and ultraviolet spectrophotometry to determine Polygonum orientale raw herbs and Polygonum orientale chemicals group quality control standards. Cell cycle and apoptosis of human hepatoma SMMC-7721 cells using flow cytometry of Polygonum orientale effective chemicals group, H22 solid tumor HE staining were observed, Polygonum orientale liver cancerous mechanism were studied. Results: 1. Polygonum orientale taxifolin, quercetin and a cream rate of indicators to determine the optimum extraction of Polygonum orientale against liver cancer component: 6 times the amount of 60% ethanol was heated to reflux extracted three times, each time for 2 h. (2) to determine the the purified Polygonum orientale chemicals group experiment for HPD-300 resin, dynamic elution conditions on the sample pH = 4 for sample concentration of 0.4 g / ml, with 70% ethanol to 2 mL / min The speed of elution, fractions were collected, vacuum drying, or of Polygonum orientale effective chemicals group. Polygonum orientale chemicals group 0.125 1 mg / mL concentration range of human hepatocellular carcinoma SMMC-7721, HepG2 cells could inhibit the proliferation inhibition rate increased with increasing dose, showed a good dose - time - effect relationship; different dose groups Polygonum orientale effective chemicals group containing serum inhibited the proliferation of HepG2 and SMMC-7721 cells, also found a good dose - time - effect relationship. Polygonum orientale anti-hepatoma effective substance group effect on the human normal liver cell toxicity rating of 1-2 in each group did not differ significantly. Polygonum orientale raw herbs and Polygonum orientale chemicals group quality control standards compliance test. Polygonum orientale chemicals group MTD equivalent to 244 times the clinical daily dose for adults. Polygonum orientale chemicals group three dose groups H22 solid tumor in mice has an inhibitory effect on the inhibition rates were 43.82%, 49.87%, 52.10%. Flow cytometry (FCM) detection Polygonum orientale chemicals group on human hepatoma cell SMMC-7721 cell cycle, different concentrations of Polygonum orientale chemicals group can cause the SMMC-7721 cells S phase arrest phenomenon, thus preventing the cells into the G2 / M phase, into the G0/G1 phase of the cell had an impact. Double staining Annexin V-EGFP/PI and Polygonum orientale effective chemicals group on SMMC-772l apoptosis, Polygonum orientale chemicals group can induce apoptosis of SMMC-7721 cells, and this induction effect has evidence of a dose-response relationship. Conclusion: Polygonum orientale optimum extraction screening stable, viable, this experiment to take advantage of this medicinal provides basic information. 2 macroporous resin better purification effect of anti-cancer compounds in Polygonum orientale, HPD-300 Polygonum orientale purified a strong anti-liver-tumor activity. 3 in vitro and in vivo efficacy experiments prove of Polygonum orientale effective chemicals group has a good anti-liver tumors. Verified for the first time Polygonum orientale chemicals group the pharmacological effects of anti-liver tumor. 4 by flow cytometry for the first time a preliminary study on the impact of Polygonum orientale on the liver tumor cell cycle, anti-tumor mechanism of action of Polygonum orientale.

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