Study On The Extraction And Separation Of Effective Parts From The Centipedesmall Molecule Polypeptide And Screening The Activity Of Antitumor

Objective Research the optimum technological conditions of centipede’s extraction by trypsin enzyme solution and Test antitumor polypeptides activity of more effective parts by Separation and purification Methods:1. Adopt L9 (34) orthogonal design method, Take pancreatic enzyme dosage, temperature and the time of extraction as three factors, and each takes three levels.The best extraction process conditions are selected by the index of extraction rate and IC5o from the MTT method of detecting various crude extractings to HepG2 cells2. Using Sephadex G-25 gel filtering chromatography to separate and purificate crude extractings from best technology.The proliferation inhibition rates of HepG2cells, Bel-7042 and A549 cells were assayed by MTT method. High performance liquid chromatographic detecte the composition of the most effective part.The HepG2 cells apoptosis were detected by Hoechst fluorescent staining. Cell cycle were detected by flow cytometry. Results:1. The optimum technological conditions of trypsin enzyme solution for: 2.0 g centipede superfine powder, the does of pancreatic enzyme is 0.1 g, the temperature of extraction is 46℃, the time of extraction is 5h2. getting three parts by the sephadex-25 gel filtration chromatography The second parts have the Strongest killer role for HepG2 cells、Bel-7042 and A549 cells.3. Compared to the negative control group, under the effects of effective component the cell cycle representation shows that the cell proportion decrease at G0/G1 period and increase at G2/M, which has statistical significance (P<0.01)Conclusions:1. This study validate that using enzyme hydrolysis and verified gel chromatography to separate and purificate effective parts of process extraction centipede small molecule polypeptide is feasibility2. The effective parts A2 of centipede small molecules polypeptide has ability of anti-tumor activity in vitro remarkably3. The effective parts A2 of centipede small molecules polypeptide can cause cell apoptosis. The main effects is to maintain cells at G2/M period, reducing mitosis and thus inhibiting the growth of hepatocarcinoma.Key words:Centipede; Extraction process; Orthogonal design; Purification; Antitumor...