[Correlation between circRNA0003353 in Peripheral Blood Mononuclear Cells and Immune Inflammation in Rheumatoid Arthritis Patients with Damp Heat Obstruction Syndrome].
To investigate the expression of circRNA 0003353 in the peripheral blood mononuclear cells (PBMCs) of rheumatoid arthritis (RA) patients with dampness heat obstruction syndrome and to examine its effect on inflammatory response of fibroblast-like synoviocytes (FLS).
The PBMCs and serum samples of 55 RA patients with dampness heat obstruction syndrome and 30 healthy volunteers were collected. The expression of circRNA 0003353 and its correlation with clinical indexes were examined. The circRNA 0003353 overexpression plasmid and siRNA were constructed and transfected into RA-FLS cell line. RT-qPCR was used to determine the expression of circRNA 0003353 mRNA. The expressions of interleukin (IL)-4, IL-10 and IL-17 were examined by ELISA. The expressions of Janus kinase 2 (JAK2), p-JAK2, signal transducers and activators of transcription 3 (STAT3) and p-STAT3 were exmained by Western blot. CCK-8 assay was used to assess cell viability. Cell migration was assessed with Transwell migration assay.
1) Compared with that of the normal group, the expression of circRNA 003353 in the PBMCs of RA patients with damp heat obstruction syndrome was significantly increased ( P<0.05). 2) Pearson correlation analysis showed that circRNA 0003353 was positively correlated with erythrocyte sedimentation rate (ESR), rheumatoid factor (RF), receptor activator of nuclear factor-κ B ligand (RANKL) and DAS28, and circRNA 0003353 was negatively correlated with IL-10 ( P<0.05). 3) The findings on the association patterns showed that the increase in circRNA 0003353 was significantly correlated with the increase of ESR, IL-17, CRP and immunoglobulin (Ig) G. 4) Logistic regression analysis showed that circRNA 0003353 was a risk factor for RANKL, CRP and ESR. 5) RT-qPCR results showed that the expression of circRNA 003353 mRNA in pcDNA3.1-circRNA 0003353 group was significantly higher than that in pcDNA3.1-NC group ( P<0.05), and that the expression of circRNA 003353 mRNA in si-circRNA 0003353 group was significantly lower than that in si-NC group ( P<0.05). 6) ELISA and Western blot results showed that, compared with those of pcDNA3.1-NC group, the expression of IL-10 in pcDNA3.1-circRNA 0003353 group significantly decreased, the expression of IL-17 increased, and p-JAK2/JAK2 and p-STAT3/STAT3 ratios significantly increased ( P<0.05). Compared with those of si-NC group, the expression of IL-10 in si-circRNA 0003353 group significantly increased, the expression of IL-17 and JAK2 decreased, and p-JAK2/JAK2 and p-STAT3/STAT3 ratios significantly decreased ( P<0.05). 7) The results of CCK-8 and Transwell assays showed that the viability and migration of RA-FLS in pcDNA3.1-circRNA 0003353 group were higher than those in pcDNA3.1-NC group ( P<0.05). Compared with those of si-NC group, the viability and migration ability of RA-FLS in si-circRNA 0003353 group decreased ( P<0.05).
The expression of circRNA 0003353 is up-regulated in RA patients with damp heat obstruction syndrome, and it is involved in the pathogenesis of RA by activating the JAK2/STAT3 signaling pathway and promoting the inflammatory response.
Wang J
，Liu J
，Wen JT
，Wang X
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Triptolide decreases rheumatoid arthritis fibroblast-like synoviocyte proliferation, invasion, inflammation and presents a therapeutic effect in collagen-induced arthritis rats via inactivating lncRNA RP11-83J16.1 mediated URI1 and β-catenin signaling.
Our previous study observed that long non-coding RNA (lncRNA) RP11-83J16.1 promoted rheumatoid arthritis (RA)-fibroblast-like synoviocyte (RA-FLS) proliferation, invasion and inflammation, which was downregulated by triptolide treatment. Therefore, the present study aimed to further investigate the mechanism and interaction between triptolide and lncRNA RP11-83J16.1 in RA treatment in vitro and in vivo.
RA-FLS was isolated and treated by different concentration of triptolide and lncRNA RP11-83J16.1 overexpression plasmid. Furthermore, collagen-induced arthritis (CIA) rat model was constructed followed by triptolide and lncRNA RP11-83J16.1 overexpression plasmid treatment.
Triptolide inhibited RA-FLS viability and lncRNA RP11-83J16.1 expression in a dose-dependent manner. Afterward, triptolide treatment inhibited RA-FLS proliferation, invasion, levels of inflammatory markers (TNF-α, IL-1β, IL-6, MMP-3, and MMP-9), inactivated lncRNA RP11-83J16.1, URI1 and β-catenin signaling, but promoted apoptosis. However, lncRNA RP11-83J16.1 overexpression weakened the effects of triptolide on regulating RA-FLS cell behaviors, URI1 signaling and β-catenin signaling. In CIA model, triptolide decreased arthritis score, hyperproliferation of synovial cells, inflammation infiltration of synovial tissue, inflammatory markers (TNF-α, IL-1β, IL-6, MMP-3, and MMP-9), inactivated lncRNA RP11-83J16.1, URI1 and β-catenin signaling, but increased cell apoptosis rate of synovial tissue. Nevertheless, lncRNA RP11-83J16.1 curtailed the treatment effect of triptolide in CIA model.
Triptolide decreases RA-FLS proliferation, invasion, inflammation and presents a therapeutic effect in CIA model via inactivating lncRNA RP11-83J16.1 mediated URI1 and β-catenin signaling.
Piao X
，Zhou J
，Xue L
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Triptolide promotes the apoptosis and attenuates the inflammation of fibroblast-like synoviocytes in rheumatoid arthritis by down-regulating lncRNA ENST00000619282.
Rheumatoid arthritis (RA), recognized as a common chronic autoimmune disease, is characterized by the excessive proliferation and inflammatory infiltration of fibroblast-like synoviocytes (FLS). In this study, our purpose is to elucidate the mechanisms of triptolide (TPL) in the treatment of RA by regulating the long non-coding RNA (lncRNA) ENST00000619282, which promoted apoptosis and reduced inflammatory infiltration of FLS in RA (RA-FLS). RA-FLS was treated with different concentrations of TPL at different time points. CCK-8 assay, ELISA, RT-qPCR, immunofluorescence, TUNEL assay, and the transmission electron microscopy were used to measure the changes of cell viability, apoptosis, and the release of inflammatory cytokines. Next, the involvement of ENST00000619282 in TPL-mediated protection against RA was explored. ENST00000619282 expression was significantly increased in the peripheral blood mononuclear cells (PBMCs) of RA patients. ENST0000061928 expression in RA PBMCs was positively associated with ESR, RF, CCP, and DAS28, while TPL treatment led to a downregulation of ENST00000619282. In addition, ENST00000619282 was significantly increased in RA-FLS. Furthermore, overexpression of ENST00000619282 elevated the levels of pro-apoptotic and pro-inflammatory factors, while reduced the levels of anti-apoptotic proteins and antiinflammatory factors. Besides, TPL treatment could reverse these effects by ENST00000619282 overexpression. The anti-RA potential of TPL might be achieved by downregulating ENST00000619282, thereby promoting apoptosis, and reducing the inflammatory response in RA.
Wen J
，Liu J
，Wang X
，Wang J
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