The Extration And Separation Of Compound From Natural Polygnum And The Antitumor Effect Detection

Objective To extract and isolate small natural compounds from fresh Polygonum by classical methods and screen their anti-tumor activity.Methods (1) The experiments on the water extract and the ethal actate extract of this plant were done by systemic solvent extraction,column chromatographic methods such as silica gel, polyamide, Sephadex LH-20 and recrystallization. And then the structure of the compounds were elucidated on the basis chemical evidence and spectral analysis.(2)The cells growth inhibition rate of different concentrations of emodin, epicatechin, stilbene glycosides and gallic acid were detected by MTT assay on SKOV-3, and the inhibitory activity of the drug were then determined. (3)The inhibition of the proliferation and the induction of apoptosis of emodin, epicatechin, gallic acid were observe by HE staining. (4) The poptosis induced by epicatechin, gallic acid on SKOV-3 cells were detected by Annexin V-FITC kit. (5) The synergistic antitumor effection of gallic acid and cisplatin, gallic acid and sodium butyrate on SKOV-3 cells were detected by MTT assay.Results 1. Nine compounds were isolated from the water extract and the ethal actate extract of fresh Polygonum, of which four were identified by spec-tral analysis. They are emodin, epicatechin, stilbene glucoside and gallic acid.2. The results of MTT assay tests showed that gallic acid inhibits the SKOV-3 cells more sufficient than the other compounds and the half maximal inhibitory concentration(IC50) for the gallic acid is 23.4μg.ml-1. When given the gallic acid at the concentration of lower than 8.5μg.ml-1,there was no cytotoxi-city for SKOV-3 cells (Inhibitory ration is lower than 5%).3. The cytoplasmic concentration, deep-dyed of the SKOV-3 cell and significant inhibiton of cell proliferation can be observed by HE staining at the half maximal inhibitory concentration(IC50) of the emodin, epicatechin, gallic acid.4. The apoptosis of SKOV-3 induced by epicatechin, gallic acid can be observed by Inverted fluorescence microscope under the Annexin V-FITC staining kit in their half maximal inhibitory concentration.5. Even in a noncytotoxic concentration, gallic acid could efficiently increase the cell inhibition rate of Cisplatin. And it showed simple adduct effect when combinated with Butyric acid Sodium.Conclusions 1. Nine compounds are isolated from fresh Polygonum and some of them have a certain degree of anti-tumor effects, Such as emodin, epicatechin, and gallic acid.2. Gallic acid can efficiently inhibite the cell growth when combinated with Cisplatin even in a non-cytotoxic concentration, which indicates that there is potential research value on the study of the increasing antitumor effects of clinical chemotherapy drug Cisplatin in future.