| Panax ginseng C. A. Mey, as valuable medicinal herbs, has a history of drug use for thousands of years. Ginsenoside is the main active ingredient and how to get more ginsenoside has become a key, microbial transformation method due to the simple culture, more variety, low cost, less deputy product to become the commonly used method of transforming ginsenoside. Whether the conversion products of ginseng by Bacillus polymyxa transformation can enhance the anti-tumor activity and what is its anti-tumor mechanism and the material basis is to be further studied. Therefore, this paper aims to study the antitumor activity and mechanism of transformed ginseng products by Bacillus polymyxa in vivo and in vitro and explores the variation of ginsenoside of the material basis of the anti-tumor effect and isolate and identify the transformed ginsenoside enzyme in Paenibacillus polymyxa to provide a theoretical basis for antitumor research and application ginseng of products by microbial transformation for future. The results were as follows:1. For study on the antitumor activity of the conversion products of ginseng by Bacillus polymyxa transformation in vitro, the conversion products had better anti-tumor effect compared with gingseng group(P < 0.05) and its IC50 value of human breast cancer cell line MCF-7 and human skin cancer A375 are 130.08 μg/m L and 339.41 μg/m L, respectively. Cell scratch test showed that conversion products could inhibit the biological information communication between cells. Flow cytometry showed that apoptosis mainly occurred in the late stage, while the early stage was less.2. For preliminary exploration of the antitumor activity and mechanism of the conversion products of ginseng by Bacillus polymyxa transformation in vivo, Compared with the model group, the low, medium and high dose group of the conversion products of ginseng by Bacillus polymyxa could inhibit the growth of H22 Tumor bearing mice, improved the organ index of mice(P < 0.05), protected the complete morphology of splenic tissue, promoted the necrosis of the tumor tissue, promoted the tumor apoptosis of the mice bearing H22(P < 0.05), increased the activity of immune cells, CD4/CD8 ratio, improved the contents of Ig G, Ig M, Ig E and the levels of cytokines TNF-α, IFN-β, IL-2, IL-6, IL-12 in serum and decreased TGF beta content that can inhibit IL-2 in serum. The difference between the ginseng group, raw powder group of ginseng and the low dose group were not significant but the middle and high dose group had significant difference(p<0.05). There was no significant difference between the model group and the bacterial fluid group. Positive drug 5-FU had a significant inhibitory effect on tumor(P<0.05) but 5-FU also reduced the immune organ index, spleen lymphocyte proliferation activity and had some side effects.3. For study on the antitumor material basis of the conversion products of ginseng by Bacillus polymyxa transformation, the contents of ginsenoside Rb1 and Rd in conversion products increased significantly(P<0.05). In the transformation of ginsenoside monomer, the ginsenoside Rc was partly converted to ginsenoside Rd.4 For the isolation and identification of ginsenoside transformation enzyme, a molecular quantum about 20 k D protease was obtained by the N terminal amino acid sequencing to identify the purified substance, the enzyme was regarded as hypothesis protease through NCBI blastx and we speculated that the enzyme could be related to ginsenoside transformation. |
PDF Full Text Request