LncRNA-ANRIL/miR-34a-mediated polarization of rheumatoid arthritis macrophages to regulate the adhesion, morphology, and survival of fibroblasts

LncRNA-ANRIL/miR-34a介导的类风湿性关节炎巨噬细胞极化调节成纤维细胞的粘附、形态和存活

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Abstract

BACKGROUND: This work focused on exploring the regulatory effect of long non-coding RNA (lncRNA)-ANRIL/miR-34a on rheumatoid arthritis (RA) fibroblasts (FLS), aiming to provide experimental support for clinical treatment of RA. METHODS: The pcDNA, pcDNA-ANRIL, antagomiRNA, antagomiR-34a, pcDNA-ANRIL, agomiRNA, pcDNA-ANRIL, and agomiR-34a were transfected into MH7A cells using liposomes. M1 macrophages and MH7A cells were co-cultured in Transwell, and the proliferation of MH7A cells at different time points was observed using crystal violet staining (CVS). The effect of M1 macrophages on the proliferation of MH7A cells was detected by MTS method. The expressions of ANRIL and miR-34a in MH7A cells were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The apoptosis rate was detected by flow cytometry (FCM). The protein expressions of Caspase-3, Bcl-2, and Bax were detected by Western blotting (WB). RESULTS: Compared with the HFLS group, the ANRIL in the MH7A group was increased (P < 0.05). After overexpression of ANRIL, the apoptosis rate (AR) of MH7A cells was markedly decreased, the expressions of Caspase-3 and Bax decreased, and the expressions of Bcl-2 increased (all P < 0.05). MH7A cell proliferation was significantly inhibited in M1 macrophage co-cultures. Following miR-34a inhibition, MH7A cells exhibited an anti-apoptotic effect comparable to that observed with ANRIL overexpression. CONCLUSION: M1 macrophages in RA joint synovial tissue could inhibit the proliferation of MH7A cells. LncRNA-ANRIL was highly expressed in RA synovial cells (SCs) and could inhibit the apoptosis of RA SCs, which was correlated with targeting miR-34a, inhibiting apoptosis in anti-RA SCs. CLINICAL TRIAL NUMBER: Not applicable.

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