AB0062 TIME COURSE OF INTESTINAL PERMEABILITY AND BACTERIAL TRANSLOCATION IN THE MODEL OF ADJUVANT-INDUCED ARTHRITIS

Abstract

Background:Intestinal inflammation, dysbiosis, intestinal permeability (IP) and bacterial translocation (BT) have been identified in patients with spondyloarthritis but the time at which they appear and their contribution to the pathogenesis of the disease is still a matter of debate.Objectives:To investigate the time-course of intestinal inflammation, IP and BT in a rat model of reactive arthritis, a subgroup of SpA, the adjuvant-induced arthritis model.Methods:Adjuvant-induced arthritis (AIA) was induced in 6-week-old male Lewis rats by an injection at the base of the tail of Mycobacterium butyricum with incomplete Freund’s adjuvant (Day (D) 0). Control rats received saline using the same procedure. Body weights and a clinical arthritis score were daily assessed. A group of AIA and control rats (n=15 per group) were euthanized at three different times of arthritis: D4 for the pre-arthritic phase (AIA-preclinical), D11 for the onset of arthritis (AIA-onset) and D28 for the acute phase (AIA-acute). In each group (AIA and control, n=15 per group)), IP was assessed by measuring plasma levels of zonulin (ELISA) and ileal mRNA expression of zonulin and occludin (RT-qPCR), BT was studied by measuring bacterial endotoxins (or LPS, by LCMS2 method), soluble CD-14 (sCD14, ELISA) and ileal mRNA expression of TLR-4, and intestinal inflammation was assessed by measuring ileal mRNA expression of IL-8, IL-33, IL-17, IL-23p19 and TNF-α (RT-qPCR). Joint damage was assessed by the determination of a clinical and radiographic score of hind paws.Results:Body weights of AIA rats decreased from D4 to D28 as compared to controls, in parallel to the development of a severe clinical and radiographic arthritic disease from D11 and D28. Compared to control rats, AIA induced an increase in plasma zonulin levels at D4, D11 but not at D28. Ileal mRNA zonulin overexpression occurred at D11 while occludin was unchanged. As early as Day 4 (preclinical phase), mRNA of IL-8, IL-33 and IL-17 were overexpressed in ileum from AIA. At Day 11 (onset), overexpression of IL-8 persisted and mRNA of TNF-α and IL-23p19 increased in AIA. Neither LPS levels nor ileal mRNA expression of TLR-4 were changed by arthritis whatever the phase of arthritis. By contrast, blood levels of sCD-14 was significantly increased in the AIA group at all stages of arthritis. No correlation was found between clinical and radiographic arthritis scores and zonulin or LPS levels. Conversely, a negative correlation was observed between intestinal IL-8 mRNA expression and arthritis score (r=-0.3, p=0.02).Conclusion:In an animal model of SpA, intestinal inflammation and increased intestinal permeability occur prior to joint inflammation, suggesting a role of these disorders in the pathogenesis of this disease.Acknowledgements:I would like to thank the Société Française de Rhumatologie for its support in this work.Disclosure of Interests:None declared