| Allium chinense is Allium genus plants in Liliaceae family, which is also called Allium chinenes G. Don., Allium Macrostemon Bunge. Allium chinense is not only edible as vegetable, but also it is as traditional medicine to treat many diseases. So to study the biological activity materials has important significance to do the high threshold processing utilization and deep development. The research about the saponins compounds was a important aspect. Consequently, the current major topic was that to research the function and mechanism of antibacterial and antitumor on Allium chinense saponins. The main research results were as follows.1. Content determination, extraction condition optimization and purificat-ion technology of ACSDetermining the content of ACS with vanillin-sulphuric acid colorim-etric, at the same time, we used dioscin as the reference substance to draw standard curve and determined the linear regression equation. And then to examine the accuracy of the experiment results by a series of experiments, and the results showed that the linear regression equation of dioscin was Y=3.9094X+0.0085. Dioscin and ACS had excellent stability color to determine between 30min and 50min. The average plus sample recovery of ACS was 94.8%. And the best extraction technology was that 60% alcohol was as solvent, materials liquid ratio was 1:12, supersonic time was 50 min, temperature was 50℃ and pH value was 5. And we used macroporous adsorption resin to purify the ACS and investigated the factors which effected the purification of macroporous adsorption resin to ACS, the results showed that D101 macroporous adsorption resin had a good adsorption and desorption capability. The maximum static adsorption ratio was 94.33%. The maximum desorption rate was 92.69% in 10h. and used 70% ethanol to elute the ACS which had the highest elution rate et al. Moderately increasing velocity could improve the elute rate for ACS.2. Isolation and the structure identification of the ACSThe ACS was isolated and purified with reversed phase high performance liquid chromatography, as a result, eight kinds of saponins were isolated and with the help of mass spectrometry to appraisal and explained the molecular weight and structures.3. The antimicrobial activity and its mechanism of ACSSaponins was tested on different microbials. The antimicrobial activity of total saponins was determined with filter slice method and studied its antimicrobial effect mechanism from physiological metabolism aspects. The results showed that the saponins had excellent antimicrobial activity to the Saccharomycete, Tritirachium album and Staphylococcus aureus, but according to the Escherichia coli., Bacillus sublitis and Pseudomonas aeruginosa, which had very weak or not inhibition action. The saponins,30% and 60% ethanol crude extraction had a significant impact to physiologica metabolism of three kinds of fungus, for example, the ability of using glucose, growth velocity and catalase viability of three kinds of fungus all were decreased. at the same time, protein content determination with Brandford method and SDS-PAGE showed that the protein contents were reduced and some protein spectrums were weakened obviously even disappeared when Tritirachium album Staphylococcus aureus and Saccharomycete were treated with saponins. So the antimicro-bial mechanism of saponins was that it reduced the use glucose efficience of microbials, then affected their growth and proliferation, reduced the viability of some key enzymes in physiological metabolism, and further to suppress the synthesis of related protein, eventually, it played the antibacterial effect.4. The anticancer activity and its mechanism of ACSB16 cells and 4T1 cells were treated with ACS and the results showed that the cells and nucleus morphology and structure changed dramatically, cytoskeleton destroyed and mitochondrial membranes potential collapsed. At the same time, the results of MTT experiment showed that the ACS had reduced cell activity, destroyed the biological membranes structure and decreased cell proliferation and migration ability, hydrogen peroxide activity and made DNA generate irregular fragment, The cell colony forming assay was showed that it indicated the ACS has obviously effect to inhibited the cell cloning formation and blocked at G0/G1 phase and made them into S period and G2/M period cells reduced with the help of Flow cytometry. Which also promoted Caspase-3 to be expressed and activated. Then established mouse tumor-burdened model to research the antitumor function of ACS in vivo, the results showed that ACS has obvious inhibited growth of tumors and a certain of protective action to the spleen and liver. |
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