| Backgrounds and ObjectiveIn the last decade, the gut microbiota has stimulated interest in the larger growing fields of microbial ecology, medicine, genomics and others. Our health not only depends on host genetic factors, but also the microbiota of human.The NIH Common Fund Human Microbiome Project(HMP) is developing research resources to enable the study of the microbial communities that live in and on our bodies and the roles they play in human health and disease. Tradition Chinese Medicine have some advantages of steady therapeutic effect on Intestinal diseases and the effect of some Chines herb may enrich the amounts of beneficial bacteria and to regulate the gut microbiota. It is difficult to change a person’s genes, but the structure changes of gut microbiota in human body is relatively easy, so the gut microbiota is an ideal drug targets. There are a large number of studies have shown that Chinese herbal can regulate the gut microbiota and help to treat some chronic diseases like type 2 diabetes, obesity, IBD and so on.Hemp seed has been documented as a folk source of food throughout recorded history, raw, cooked, or roasted, and hemp seed oil(HSO) has been used as a food/medicine in China for at least 3000 years.In our studies, firstly, to establish GC method for determination of oleic acid, linoleic acid and α-linolenic in the HSO. Secondly, to study the effects of hemp seed on the gut microbial community structure and Intestinal immunity function in mice.Content and results1.To establish a GC method for determination the content of oleic acid, linoleic acid and α-linolenic in the hemp seed oil. GC was performed on the CP-WAX 52CB(30 m×0.32 mm, 0.50μm)capillary column at 205 ℃, the detector was FID at 300 ℃, the injector temperature was 250 ℃, the split ratio was 10:1, and 1μL was injected. Results The linear range of methyl oleate, methyl linoleate and methyl α-linoleic had good linear relationship in their concentration range of 7.6 ~ 152 μg·m L-1(r=0.9999), 8.0 ~ 160μg·m L-1(r=0.9997)and 9.7~ 194 μg· m L-1(r=0.9998),and the average recover were96.55 %, 99.68 % and 102.22 %; RSD were 1.35 %, 2.79 % and 1.19 % respectively. It has 29.36 mg oleic acid, 137 mg linoleic acid and 55.31 mg α-linoleic acid per gram of the hemp seed. This method was sensitive, accurate and reproducible, which may provide the basis for the quality evaluation and control of oleic, linoleic acid and α-linoleic acid of the hemp seed oil.2.To find out whether hemp seed water extract modulates the composition of the gut microbiota during treatment. Bacterial DNA was extracted from the fecal and characterized by analysis of the hypervariable V4 fegion of the 16 S r RNA. It turned out that the sequencing data has good quality, thirteen bacterial phyla were identified. Most belonged to four phyla: Bacteriodetes, Firmicutes and Proteobacteria. We were able to get an average of82,609 gene sequences per sample.A significant increase in the proportion of phylum Bacteroidetes species and decrease in the proportion of phylum Firmicutes species were observed for mice after treatment. So the hemp seed water extract may have effect on modulating the composition of the gut microbiota.3.To investigate the change in content of intestinal mucosal SIg A of mice with hemp seed water extract. The results showed that SIg A contents were significantly increased in hemp seed water extract low, middle and high dose groups compared to control group. It was concluded that hemp seed water extract have effect on intestinal mucosal immune system, but further research is needed to understand the molecular mechanisms.4. Blood was sampled from eyeball for separating the serum under sterile condition,the cytokines levels of IL-4, IL-10, IL-17, IFN-γ were detected by ELISA. The experiment results show that there were no significant difference in the levels of IL-4 and IL-17 between the control group and hemp seed water extract group. Compare with control group,the levels of IL-10 and IFN-γ were significantly increase in hemp seed water extract highdose group that indicated hemp seed water extract was involved in the inflammation through the regulation of IL-10 and IFN-γ levels in mice.The results suggest that: the hempseed benefited regulating the intestinal microbiota and studying the mechanism of interaction. |
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