| Objective:To investigate whether Kuijie enema Ⅱ(Ke Ⅱ)can adjust the intestinal flora,increase the content of short-chain fatty acids(SCFAs),balance the TH17/Treg immune axis,and thus achieve the effect of treating UC.Methods:Using 2.5%DSS solution to induce replication of UC mouse model,and randomly divided into three groups of normal group,model group and drug administration group,8 in each group;daily observation of the weight loss rate,stool texture and bleeding of each group of mice Calculate the disease activity index;record the length of the colon on the day of sacrifice.Pathological sections of colon tissue were prepared,and the pathological and morphological changes of colon tissue were evaluated by light microscope;flow cytometry was used to detect the number of Treg and Th17 cells in mesenteric lymph nodes of mice;real-time quantitative PCR(qPCR)was used to determine the inflammatory factor IL-in mouse colon tissue 17.The expression levels of IL-10 and transcription factors RoRγt and Foxp3.Using a broad-spectrum antibiotic to simulate a sterile UC mouse model,it was divided into a blank group,an antibiotic group,and an antibiotic+administration group,with 8 rats in each group,to observe the changes of the above indexes before and after administration of sterile UC mice;using 16S rRNA sequencing The changes of intestinal flora composition before and after administration of UC model mice were analyzed;the content of short-chain fatty acids(SCFAs)in feces before and after administration of UC model mice was determined by liquid chromatography tandem mass spectrometry(LC-MS/MS).Results:After DSS-induced modeling,the mice were generally in poor condition,with a significant weight loss(P<0.01)and a significantly increased DAI score(P<0.01);compared with the normal group,the pathological conditions of colon and colon tissues in the model group It showed obvious deterioration,the length of the colon was shortened(P<0.01),the intestinal flora shannon index and chaos index were significantly reduced(P<0.01),at the gate level,the abundance of Bacteroides phylum in mice was reduced,and the thick wall bacteria The degree of bacteria increased,the structure of the bacteria deviated significantly from the normal group,the number of Treg cells in mesenteric lymph nodes was significantly reduced,the number of Th17 cells increased significantly(P<0.05),the mRNA expression levels of IL-17 and RoRyt in colon tissue were significantly increased,while IL-10.The mRNA expression level of Foxp3 was significantly down-regulated(P<0.01),and the contents of 6 SCFAs in mouse feces were significantly reduced(P<0.01,P<0.05);after treatment,compared with the model group,the The general condition of the drug group improved,the pathological condition of the colon tissue improved significantly,the length of the colon increased significantly(P<0.01),the intestinal flora shannon index and chaos index increased significantly(P<0.01,P<0.05),Bacteroides at the portal level The portal abundance increased,the thick-walled bacteria portal abundance decreased,the flora structure deviated from the model group and returned to the normal group,the number of Treg cells in mesenteric lymph nodes increased significantly,the number of Th17 cells decreased significantly(P<0.05),colon tissue The mRNA expression levels of IL-17 and RoRyt were significantly down-regulated(P<0.05),while the expression levels of IL-10 and Foxp3 were up-regulated(P<0.01,P<0.05),and the contents of 6 SCFAs in mouse feces were significantly increased(P<0.01,P<0.05).In the experiment of simulating sterile UC mice,the mice were generally in poor condition after DSS modeling,their body weight decreased significantly(P<0.01),and the DAI score increased significantly(P<0.01);compared with the normal group of mice,In the antibiotic group,the pathological condition of the colon tissue of the mice deteriorated significantly,the length of the colon was significantly shortened(P<0.01),the number of Treg cells in mesenteric lymph nodes was significantly reduced(P<0.01),and the number of TH17 cells was significantly increased(P<0.01);IL in colon tissue-17.The expression level of RoRyt was significantly increased(P<0.013 P<0.05),while the expression levels of IL-10 and Foxp3 were significantly decreased(P<0.01);compared with the mice in the antibiotic group,the mice in the antibiotic+administration group were generally There was no improvement in the condition and the pathological tissue of the colon,and there was no significant change in body weight,DAI score,and colon length(ns,no significance difference).At the same time,the content of TH17 and Treg cells in mesenteric lymph nodes and IL17,IL-10,RoRyt in colon tissue,Foxp3 mRNA expression level showed no significant difference(ns,no significance difference).Conclusion:Kuijie enema Ⅱ can significantly improve the DSS-induced intestinal inflammation in UC mice,and its mechanism may be related to the regulation of intestinal flora,increase the content of SCFAs,and promote the balance of Treg/Th17 immune axis;The important target for UC in the treatment of UC is indispensable. |
