Background: It was found that the expression of mir-29b was significantly up-regulated in PBMs, and we tried to clarify TNF- α The production of proinflammatory cytokines was increased by inducing the overexpression of mir-29b in CD14 +PBMs in patients with rheumatoid arthritis (RA), and by using TNF-α that the expression of mir-29b was significantly urapy to reverse regulate mir-29b, and carry out relevant experiments to verify our scientific research hypothesis.

Objectives: By observing patients with RA treated with TNF-α significantly urapy to reverse regulate mir-29b, and carry out relevant experimenperipheral blood mononuclear cells (CD14+PBMs) and releases pro-inflammatory cytokines.

Methods: (1) Cell experiment: PBM cells from RA patients were collected and extracted for CD14+ cell expression labeling. Different doses of TNF-α blood mononuclear c500 ng/ml) were used for intervention. Meanwhile, the expression of mir-29b was analyzed by rt-qpcr at the level of TNF-α100 ng/ml for different periods of time (0,6,12,24h). In addition, the supernatant of cell culture was collected and human cytokines IL-1α, IL-1β, TNFα, IL-6, IFN-α and IL-8 were measured using a V-plex human cytokine 30-plex kit. (2)Grouping experiment of clinical intervention: 21 patients with RA diagnosis and 15 healthy volunteers were divided into three groups. TNF-α inhibitor group: RA patients treated with TNF-α inhibitor were collected (n=15); IL-6 monoclonal antibody group: RA patients treated with tocilizumab (n=6); Control group: healthy volunteers (n=15) were used as normal controls. PBMc was extracted from TNF-α inhibitor group for 6 months, IL-6 monoclonal antibody group for 6 months, and control group, respectively, to observe the difference of Mir-29b expression in CD14+PBMs of the three groups.

Results: The expression of miR-29b was dose-dependent and time-dependent with the incubation of TNF-α, and there was a significant difference (P < 0.05). Compared with the control group, the overexpression of miR-29b also led to an increase in the expression levels of a wide range of chemokines and proinflammatory cytokines (including IL-1α, IL-1β, TNFα, IL-6, IFN-α and IL-8) (P < 0.05). The expression of miR-29b in RA patients treated with TNF-α inhibitor was significantly reduced compared with that treated with Totuzumab (P<0.05).

Conclusion: TNF-α inflammatory factors can induce the overexpression of miR-29b in RA patients, and then producing a large number of proinflammatory cytokines, which can aggravate the inflammation mechanism of RA. In RA patients, TNF-α inhibitors may partially reduce the inflammatory response through the TNF-α/CD14+PBMs/ Mir-29b signaling pathway. Therefore, more attention should be paid to the expression of Mir-29b in TNF-α and CD14+PBMs in clinical practice, which may accurately indicate the state of immune disease in patients, and provide a basis for more accurate judgment of prognosis and the course of immunotherapy, as well as optimization of immunotherapy programs.

REFERENCES:

[1]Long L, Yu P, Liu Y, et al. Upregulated microRNA-155 expression in peripheral blood mononuclear cells and fibroblast-like synoviocytes in rheumatoid arthritis. Clin Dev Immunol. 2013;2013:296139.

Disclosure of Interests: None declared