Extraction, Purification, Intestinal Micoecology Regulation, Anti-Tumor And Immunoregulatory Activity Of Ciceritol From Chickpeas

Ciceritol is a trisaccharide, which is an a-d-digalactoside of pinitol and is O-a-d-galactopyranosyl-(1-6)-O-a-d-galactopyranosyl-(1-2)-1d-4-O-methyl-chiro-inositol. Chickpeas are the main resource of ciceritol, about 10% of carbonhydrates in chickpeas is ciceritol. The objective of this study is to extract ciceritol (with a purity>95%) from chickpeas, evaluate the intestinal microecology regulation of ciceritol, as well as anti-tumor and immunoregulation activity. Results are showed as follows:1 Extraction and purification of ciceritolSoluble saccharides in chickpeas was isolated by medium pressure chromatography which made with activated carbon and diatomate at weight ratio of 1:1. The following was to determine the composion of soluble saccharides by HPLC. The results demonstrated that the soluble saccharides in chickpeas were fructose, sucrose, mclibiosc, ciceritol and stachyose. The ciceritol had a purity of 95.17%.2 Study on intestinal microecology regulation of ciceritolAnaerobic faecal fermentation was applied to study the in vitro intestinal microecology regulation of ciceritol. In this study, control group was media without any carbon source, media with frctooligosaccharide was considered as positive control group. Fluorescent in situ hybridization was used to mintor the change in the population of beneficial bacterias, harmful bacterias and total bacterias. It was found that ciceritol had a negative effect on the growth of Bacteroids-Prevotella, Clostridium histolyticum and Eubacterium-Clostridium groups, however, a positive effect on the growth of Bacteroids-Prevotella, Clostridium histolyticum and Eubacterium-Clostridium groups was observed. Refers to total bacterias, the population didn’t affected by ciceritol. The concentraton of short-chain fatty acid (SCFA) was measured at different incubation time. Compared with control group, the concentration of SCFA in ciceritol group was higher after 24 h incubation.3 Inhibitory effect on the proliferation of HGC-27 cellMTT assay was applied to evaluate the inhibitory activity of ciceritol for the proferation of HGC-27. The results demonstrated that the inhibitory activity was enhanced with the increase of incubation time, as well as the concentration of ciceritol. After 72 h incubation,82% of inhibitaion was obtained.4 Immunoregulatory activity of ciceritolFor the immunomodulatory activity, both in vitro and in vivo evaluation were preformed. Creation of animal model was used for in vivo evaluation. The results demonstrated that the index of spleen was increased with the supplement of ciceritol, the same phenomenon was observed for the swelling rate of ear, as well as the hemolysin levels.Cell model was applied to in vitro experiment. It was found that ciceritol could improve the proliferation of RAW264.7, as well as the release of NO. For the immune reactive molecules (IL-6、IL-1β、IFN-α andIFN-γ), ciceritol had a positive effect with a concentration of 200μg/mL...