Chemical Composition Studies And Screening Effective Parts Of Glelitsia Sinensis Lam Extract On Anti-liver Cancer

Glelitsia(Glelitsia sinensis Lam) is the common treatment Combination drugs such as breast cancer,lung cancer,liver cancer. The anti-tumor test and clinical study confirmed the total extract Gleditsia has anti-cancer effects.This experiment was based on the total extract of Gleditsia had effective anti-cancer and Mechanism of the initial research.use of modern separation method, To separation The total extract of Gleditsia and screening of anti-tumor. To determine the Anti-tumor activity parts of Gleditsia. Carried out extraction,separation, and purification for determining the effective parts of the Glelitsia. And get compounds,and carry on structural identification. Through this study, further analysis of traditional Chinese medicine for the anticancer activity of Gleditsia provide the basis of material 1.In this study, by consulting the relevant literature, Combined with pre-extraction conditions on the honey locust compared, Ultimately determine the total extract of Gleditsia the best extraction technology:Gleditsia Pharmacognosy drying,grinding, sieving through 40 mesh. Were sampled by solid-liquid ratio 1:10, 1:8,1:875% ethanol solution reflux extract three times.Extraction time was 3h,2h,2h. Combined extract,Evaported,The total extract obtained.1.Using the system solvent divided total extract Gleditsia into several different parts of polar extract.through animal vivo Pharmacodynamics testing,Screening for pharmacological activity from Gleditsia petroleum ether part,Ether part,Ethyl acetate part,and N-butanol part.The results show that,Gleditsia ethyl acetate part has significant pharmacological of anti-tumor activity.2.By in vitro cell test,Detection of Gleditsia extract three different concentration groups(high,medium,low) On be1-7402 human hepatocellular carcinoma cell proliferation,cell cycle.Liver cancer related genes(bax,bcl-2,p53)expression and telomerase activity. The results showed that,Three concentrations of Gleditsia sinensis extract on human hepatoma cell bel-7402 Cell proliferation,regulation of oncogenes and tumor suppressor genes, and Promote cancer cell apoptosis,inhibit of telomerase activity differences were all have significant (P<0.05 or P<0.01).3. Gleditsia total extract adding an appropriate amount of water dilution,Using the system solvent,The total extract of Gleditsia were Separately with Petroleum ether,ethyl ether,ethyl acetate. Water-saturated butanol for extraction,Into petroleum ether,ether site,ethyl acetate extract,butanol and water parts of five different parts of the polar sites.4.By in vivo testing in animal Gleditsia petroleum ether,ether site,ethyl acetate extract,butanol site of screening for pharmacological activity. The results show that,Gleditsia parts of ethyl acetate extract and butanol has significant antitumor activity management.5.Ethyl acetate extract of the Gleditsia mechanism of inhibition of liver cancer cells for further study,Found that high-dose group Gleditsia parts of ethyl acetate in mice liver cells by enhancing the Smad4 gene and protein expression and reduced expression of Smad7 gene and protein,So as to achieve the role of inhibition of tumor cell proliferation.6.Ethyl acetate extract of the Gleditsia and butanol sites were separated by silica gel column chromatography,use 100-200 and 200-300 respectively present silica gel column chromatography to separate projects.After repeated separation,Recrystallization. Been successful compounds.7.By1H-NMR spectroscopy,1H-1H COSY spectrum,13C-1H COSY spectrum for structural identification of compounds derived from. Structure derived compounds successfully.1.The optimum extraction of this experiment is a simple technique, efficiency of extracting effective components,extracted completely.Help extract the maximum quantity of active ingredients in medicinal herbs.2.On the extraction of Gleditsia extract to in vitro pharmacological experiments,Experimental results show,Gleditsia sinensis extract on human hepatoma cell bcl-740 significantly inhibited the proliferation of,Promote apoptosis.3.By Pharmacological screening of animal experiments,Gleditsia ethyl acetate extract and butanol extracts of human liver cancer cell be-7402 significantly inhibited proliferation(P<0.01). Identified the ethyl acetate extract of Gleditsia extract and butanol extract have the exact anti-tumor activity.4.Application of ethyl acetate extract,Through the study on Mouse liver cancer cell TGF-β/Smads signal system Smad4,Smad7 gene and protein regulation of,Further clarified the role of Gleditsia extract anticancer mechanism.5.Use Silica gel column chromatography separation and nuclear magnetic resonance spectroscopy,got seven compounds,and carried out structural identification from four.Among a flavonoid, three dihydro flavonoids.Respectively: 3'-methoxy-5,7,4'-dihydroxyflavone-7-0-β-D-glucopyranoside,5,7,3',5'-tetrahydroxyflavanone-7-0-β-D-glucopyranoside,3'-methoxy-5,7,4'-dihydroxyflavanone-7-0-β-D-glucopyranoside,5,7,4'-tetrahydroxyflavanone-7-0-β-D-glucopyranoside. 6. At present,the active ingredient of anticancer Gleditsia more concentrated in the n-butanol part of saponins,the active ingredients of other studies have reported less.In this study, the active site by Gleditsia pharmacological screening,drawn parts of ethyl acetate extract and butanol all have pharmacological activity. And were separated and the components research.provide new ideas and theoretical basis for the Gleditsia anti-cancer studies.