Therapeutic Effect Of TSCS On Serum-transfer Arthritis Via Gut Microbiota Disturbance

Objective To investigate the effect of intestinal flora on mice with serum metastatic arthritis and the mechanism of total papaya glycosides in the treatment of serum metastatic arthritis.Methods（1）K/Bx N mice were born by pairing KRN males with NOD females.Blood was collected from buccal veins of K/Bx N mice,serum was separated and extracted and injected into recipient mice via tail vein.Establishment of K/Bx N mouse serum metastatic arthritis model.（2）HE staining was used to observe the ankle pathological changes in mice.（3）The positional relationship between mast cells and lymphocyte distribution was observed by immunohistochemistry.（4）The contents of inflammatory factors TNF-α,IL-6,IL-17 and PGE₂in the supernatant of mouse foot and ankle homogenate were detected by ELISA.（5）16s r DNA was used to detect the structure and quantity of mouse intestinal flora,and the correlation analysis was performed with inflammatory factors.（6）The single/combined antibiotic protocol was used to pretreat the mice to clear the intestinal flora and simulate the intestinal environment of sterile mice.Results（1）the abundance of intestinal microflora decreased in the model group,but increased in the total glycosides group and mast cell deficiency group.（2）the articular cavity was clearly seen by hematoxylin-eosin staining in the ankle joint of mice in the control group,there was no inflammatory cell infiltration in synovium and the surface of articular cartilage was smooth.The ankle joint cavity of mice in the model group almost disappeared and the infiltration of inflammatory cells was obvious.The ankle space of mice in total glucosides of Chaenomeles sinensis group and mast cell deficiency group became smaller.（3）the immunohistochemical staining of foot and ankle joint of mice with serum metastatic arthritis showed brown mast cells surrounded by light blue lymphocytes.（4）the results of detection of inflammatory factors in foot and ankle joint of mice in each group showed that IL-6,IL-17,TNF-αand prostaglandin E₂in the model group were significantly higher than those in the control group,while the contents of IL-6,IL-17,TNF-αand PGE₂in the total glycosides group and mast cell deficiency group were significantly lower than those in the model group,but there was no significant difference between the total glycosides group and the mast cell deficiency group.（5）in the histogram of community structure distribution,the number of Lactobacillus in the mast cell deficiency group was significantly lower than that in the model group（P<0.05）.The number of microbacilli in the mast cell deficiency group was significantly higher than that in the control group（P<0.05）and the model group（P<0.01）.There was a significant correlation between Lactobacillus,Helicobacter pylori,Clostridium,Microbacillus, Helicobacter pylori and inflammatory factors.（6）single antibiotic treatment can effectively remove most of the microflora in the intestinal tract of mice and simulate the aseptic state of the intestinal tract of mice,while the combination of antibiotics can only clear part of the intestinal flora of mice and cause intestinal flora disorder and accelerate the reproduction of other pathogenic bacteria.Conclusion Disturbance of intestinal flora in mice with serum metastatic arthritis induces increased levels of inflammatory factors,mast cells participate in changes in intestinal flora,causing rheumatoid arthritis.TSCS improve the intestinal microecology and regulate the intestinal flora abundance to treat arthritis;A single antibiotic pretreatment can clear most intestinal flora in mice and simulate the intestinal environment of sterile mice,while combined antibiotic treatment can aggravate the intestinal flora disturbance,and simulate the intestinal environment of sterile mice for subsequent fecal transplantation.Provides new ideas.