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.

Establishment of Thin Layer Chromatography Identification Method for Different Processed Products of Rehmanniae Radix / 中国实验方剂学杂志

Objective:To establish a thin layer chromatography （TLC） identification method for different processed products of Rehmanniae Radix. Method:Catalpol， D-fructose， sucrose， raffinose， stachyose， melibiose and manninotriose were used as control substances， and the effects of extraction solvents （water， 20% methanol， 50% methanol， 80% methanol）， developing solvents （n-butanol-methanol-chloroform-glacial acetic acid-water， ethyl acetate-pyridine-glacial acetic acid-water， n-butanol-glacial acetic acid-water）， color reagents （aniline-diphenylamine-phosphoric acid solution， ninhydrin solution）， sampling volumes （2， 4， 6 μL） and inspection conditions （sunlight， bottom lamp of sunlight， 365 nm， 254 nm） on TLC were investigated to determine the preparation method of sample solution of Rehmanniae Radix and Rehmanniae Radix Praeparata and the optimal TLC conditions. Result:High performance silica gel G plate was used for TLC， n-butanol-methanol-chloroform-glacial acetic acid-water （13∶5∶5∶1∶2） was used as developing agent， aniline-diphenylamine-phosphoric acid solution was sprayed and heated at 110 ℃ for color development， and then inspected under the bottom lamp of sunlight. The separation and color development of different processed products of Rehmanniae Radix were good with clear spots and good characteristics. Conclusion:The established TLC is simple and easy to operate with obvious qualitative characteristics and intuitive results. It can effectively identify different processed products of Rehmanniae Radix and provide experimental basis for determining the end point of processing of Rehmanniae Radix Praeparata.

Transformation and attribution of drug properties in Galla Chinesis fermented Baiyaojian based on cold and heat syndrome rats / 中国中药杂志

To reveal the transformation and attribution of drug properties in Galla Chinesis fermented Baiyaojian by studying the effect of Galla Chinesis and Baiyaojian on cold and heat syndrome rats. Euthyrox was used to induce the hyperthyrosis model,ice water stimulation was used to induce the cold syndrome model,and different concentrations of Galla Chinesis and Baiyaojian water decoction were administrated by gavage for 15 d continuously. Symptom indexes were evaluated,content of pyruvic acid( PA),ATPase activity in liver and contents of DA,T4,cAMP,5-HT,NE,17-OHCS,TRH and TSH in serum were assayed by enzyme linked immunosorbent assay and spectrophotometry. The rectal temperature,water consumption and body weight of heat syndrome rats in model group were increased,cAMP,NE,17-OHCS,TRH and PA were increased,TSH,Na-K ATPase and Ca-Mg ATPase were increased significantly( P<0. 01),while 5-HT was decreased,compared with those of the blank group( P< 0. 05),the contents of T4,DA,NE,TSH,TRH,cAMP and 17-OHCS were decreased significantly( P<0. 01),PA and Ca-Mg ATPase in WG and BG groups were decreased compared with those of the model group( P<0. 05),and the Galla Chinesis content of WG group was lower than that of BG group,while the contents of 5-HT in WG and BG groups were increased,and the Galla Chinesis content of WG group was higher than that of BG group,with no significant difference of viscera index between heat syndrome rats in blank group,model group and drug groups. The rectal temperature,water consumption and body weight of cold syndrome rats in model group were decreased,DA,T4,cAMP,NE,17-OHCS,TRH,TSH,PA,Na-K ATPase and Ca-Mg ATPase of rats in model group were decreased,whereas 5-HT was increased compared with those of the blank group( P<0. 05),the indexes of heart,lung and kidney were significantly higher than those in the blank group( P<0. 05). Both Galla Chinesis and Baiyaojian can significantly alleviate the symptoms of heat syndrome rats caused by levothyroxine sodium. Galla Chinesis has a better effect than Baiyaojian,but cannot alleviate the symptoms of cold syndrome caused by ice water stimulation,suggestting that the decoction of Galla Chinesis and Baiyaojian are both cold,but Galla Chinesis is colder than Baiyaojian. Cold property in Galla Chinesis fermented Baiyaojian can be relieved. In clinical application,the property of " slight cold" is more accurate than " neutral property" for Baiyaojian.