Abstract

BackgroundOsteoporosis(OP) is one of the major comorbidities of rheumatoid arthritis(RA) which is associated with immune disorders[1]. The gut microbiota has been highlighted to be an important environmental factor to influence immune system in maintaining bone health and regulating bone remodeling[2]. However, the alterations of intestinal flora and its relationship with immune system in RA patients with OP are unclear.ObjectivesTo investigate the characteristics of gut microbiome as well as the associations between flora and peripheral lymphocyte subpopulations and cytokines in rheumatoid arthritis patients complicated with osteoporosis.MethodsTotal 28 RA patients were divided into 14 RA-non-OP and 14 gender- and age-matched RA-OP groups according to their bone mineral density (BMD) and the history of fragility fracture. Gut microbiota of participants were investigated by 16s rRNA and peripheral lymphocyte subsets and cytokines were assessed via flow cytometry. Indicators like erythrocyte sedimentation rate (ESR), C-reaction protein (CRP), anti-cyclic citrullinated peptide antibody (ACPA) and anti-mutated citrullinated vimentin (MCV) antibody were recorded meanwhile. Alpha diversity (ACE, Chao1, Simpson, Shannon) and beta diversity indices were analyzed using QIIME2. Biomarker species were recognized based on STEMP. Spearman analysis was adopted for correlation of two variables. All P-values reported herein were two-tailed and P-value<0.05 was taken as statistically significant.ResultsThe alpha-diversity have no significant difference between RA-non-OP and RA-OP groups (P >0.05, Figure 1A). The community structure of microflora differed between two groups (P <0.05, Figure 1B). As for the composition of intestinal flora at genus level, Faecalibacterium, Proteus, Catenibacterium, Enterobacter and Erysipelatoclostridium in RA-OP group as well as Lachnospiraceae_ND3007_group, Parasutterella, Megasphaera, Tyzzerella, UCG-005, Clostridium_sensu_stricto_1, UCG-002, Lachnospiraceae_NK4A136_group, Christensenellaceae_R-7_group, Prevotella, Parabacteroides in RA-non-OP group were significantly increased (Figure 1C). There were positive correlations between Lachnospiraceae_NK4A136_group and the level of T, Th1 and Th17 cells, but negative relevance with ESR, CRP and IL-10 (P <0.05). The relative abundance of Faecalibacterium was negatively correlated with IL-2, IL-4, TNF-α and positively with MCV (P <0.05). Clostridium_sensu_stricto_1 and Lachnospiraceae_ND3007_group were negatively correlated with ACPA and MCV respectively as well as IL-2 (P <0.05, Figure 1D-E).ConclusionAbnormality of immune system may contribute directly or indirectly to OP in RA, which may be related to the disturbance of gut microbiota.

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