Analysis on Mechanism of Baiyaojian in Treating Ulcerative Colitis Based on Intestinal Flora and Intestinal Mucosal Barrier Function / 中国实验方剂学杂志

Objective:To compare the effects of Baiyaojian before and after fermentation on intestinal flora and expression of Occludin and zonula occludens protein-1 （ZO-1） in intestinal mucosa of mice with ulcerative colitis （UC）， and to explore the mechanism of Baiyaojian and Galla Chinensis in the treatment of UC. Method:Totally 50 mice were randomly divided into 5 groups with 10 mice in each group， one group was randomly selected as blank group， and the other 4 groups were treated with dextran sodium sulfate （DSS） to induce UC model. After modeling， mice in the blank group and model group were given normal saline， and treatment groups were given Mesalazine （0.8 g·kg^-1）， Galla Chinensis decoction （1.8 g·kg^-1） and Baiyaojian decoction （2.7 g·kg^-1） by intragastric administration for 7 days. The 16S rRNA sequencing technology was used to detect the changes of intestinal flora in mouse feces. The histopathological changes of colon tissue were observed by hematoxylin-eosin （HE） staining， and the expression of Occludin and ZO-1 in colon tissue of mice were compared by immunohistochemistry. Result:Compared with the blank group， the abundance and diversity of intestinal flora in UC mice were significantly decreased， and the colonic tissue was thickened with congestion and obvious ulcers， and the expression levels of Occludin and ZO-1 were significantly decreased （<italic>P</italic><0.01）. After treatment with Galla Chinensis and Baiyaojian， the abundance and diversity of flora were improved. At the phylum level， relative abundance of Firmicutes， Proteobacteria and Actinobacteria increased significantly （<italic>P</italic><0.01）， while the relative abundance of Bacteroidetes decreased significantly （<italic>P</italic><0.01） in Galla Chinensis group. In Baiyaojian group， the relative abundance of Proteobacteria and Actinobacteria increased significantly （<italic>P</italic><0.01）， while the relative abundance of Firmicutes increased and the relative abundance of Bacteroidetes decreased， but there was no significant difference. At the genus level， the relative abundance of <italic>Bacteroides</italic>， <italic>Allobaculum </italic>and <italic>Ruminococcus</italic> decreased significantly （<italic>P</italic><0.01）， the relative abundance of <italic>Roseburia</italic>， <italic>Prevotella</italic>， <italic>Oscillospira</italic> and <italic>Paraprevotella</italic> increased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01） in Galla Chinensis group. In Baiyaojian group， the relative abundance of <italic>Bacteroides</italic> and <italic>Allobaculum</italic> decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）， and the relative abundance of <italic>Prevotella</italic>， <italic>Oscillospira</italic>， <italic>Roseburia</italic> and <italic>Ruminococcus</italic> increased significantly （<italic>P</italic><0.01）. Compared with model group， colon tissue of Galla Chinensis group and Baiyaojian group was recovered obviously， congestion was alleviated， only scattered ulcers were seen. The expression of Occludin and ZO-1 increased， and the expression level of Baiyaojian group was higher than that of Galla Chinensis group. Conclusion:The effect of Baiyaojian is better than Galla Chinensis in the treatment of UC. The mechanism may be through regulating the abundance and diversity of intestinal flora， improving the disorder of intestinal flora and increasing the expression of ZO-1 and Occludin and protecting the intestinal mucosal barrier function for alleviating intestinal inflammation.

High-throughput sequencing analysis of intestinal flora diversity of two freshwater snails (Radix auricularia and Planorbella trivolvis) / 生物工程学报

Freshwater snail is an important biological group in aquatic ecosystem and an intermediate host of many parasites. Intestinal flora plays an important role in animal energy metabolism and resistance to pathogens. We analyzed the intestinal microbiota diversity of Radix auricularia (RA) and Planorbella trivolvis (PL) by 16S rRNA high-throughput sequencing. At the phylum level, RA had 23 phyla, including Proteobacteria (33.63%), Cyanobacteria (15.33%), Chloroflexi (13.95%), and Actinomycetes (12.99%). PL had 13 phyla, including Proteobacteria (54.88%), Bacteroidetes (28.49%), and Actinomycetes (7.65%). At the genus level, there were 445 genera in RA, including Pleurocapsa, Thiodictyon, Leptotrichia, and Nocardioides. There were 238 genera in PL, including Cloacibacterium, OM60NOR5_clade, Pseudomonas, and Rhodobacter. Ninety-three genera were the common core flora of the two snail species (all the samples were present), and 27 genera had an abundance greater than 0.5%. The structure of intestinal microbiota was significantly different between the two groups (P=0.027). We performed the functional prediction of intestinal microbiota using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt), and the results show that the KEGG functional composition of the intestinal flora of the two snails was similar, and the abundance of the amino acid metabolism, carbohydrate metabolism and membrane transport were large. In summary, the intestinal microbiota of the two snails was high in diversity and significantly different, but there were a large number of common core flora.

Airway Dysbacteriosis Exacerbated Murine Airway Allergic Inflammation / 中山大学学报(医学科学版)

[Objective] To investigate the effects of airway dysbacteriosis on the development of murine atlergic airway diseases (AAD).[Methods] Female C57BL/6 mice were neubulized with Vancomycin for 10 days and then were sacrificed.The bacterial population in bronchoalveolar lavage fluid (BALF) were evaluated using 16S rRNA high-throughput sequencing technology,exploriug the method of establishing an airway dysbacteriosis mouse model.After the mouse model was established successfully,airway dysbacteriosis mouse models were established by the same method,and based on that,the mice were sensitized and challenged with ovalbumin (OVA) to induce airway allergic inflammation.The frequency of nasal rubbing behaviors per mice was counted;the total cell number and eosinophil relative abundance in BALF were evaluated;the lung tissue inflammation and goblet cell metaplasia were assessed according to histopathological features;and the IgE level in serum,IFN-γ,IL-4 and IL-5 levels in BALF,and IL-33 levels in serum,BALF and intestine tissue were measured by ELISA.[Results] Nebulization of Vancomycin increased Bradyrhizobium,Sphingopyxis,Cupriavidus,Pelomonas,and decreased Akkermansia and Prevotella_6 in airway,inducing significant airway dysbacteriosis.Using the animal model,further study found that airway dysbacteriosis exacerbated OVA-induced airway allergic inflammation,including increased nasal rubbing frequency,higher serun IgE level,more total cell count especially eosinophil infiltration,more serious lung tissue inflammation and goblet cell metaplasia.Additionally,compared to OVA group,mice in Dysbacteriosis and OVA group had significantly increased level of Th2 cytokine IL-4 and IL-5,and significantly decreased Thl cytokine IFN-γin BALF,which revealed that mice in Dysbacteriosis and OVA group had mote remarkable Thl/Th2 imbalance.Furthermore,IL-33 level showed a significant increase in BALF,but didn't change in serum or intestine tissue in Dysbacteriosis and OVA group compared to OVA group.Indicating that airway dysbacteriosis may only affect the local production of IL-33.[Conclusions] An airway dysbacteriosis mouse model was established by Vancomycin nebulization successfully.Airway dysbacteriosis may activate innate lymphoid cells (ILC) and Th2 cell by inducing local IL-33 secreting,which leads to the imbalance of Th1/Th2,and in turn promotes the development of AAD.