Department of Orthopedics, Tianjin First Central Hospital, No.24 FuKang Road, Nankai District, Tianjin, 300192, China.
Department of Orthopedics, Tianjin Medical University General Hospital, No.154 Anshan Road, Heping District, Tianjin, 300052, China.
Inflammation. 2021 Aug;44(4):1359-1369. doi: 10.1007/s10753-021-01421-2. Epub 2021 Feb 6.
Rheumatoid arthritis (RA) is a common chronic autoimmune disease featured by synovial inflammation. miR-496 is closely involved in various pathologic conditions. However, its role in RA has not yet been elucidated. Expression of miR-496 and MMP10 was determined based on the clinical samples with RA retrieved from the Gene Expression Omnibus (GEO) datasets. In vitro model of RA was constructed in MH7A cells stimulated by IL-1β (10 ng/mL). Cell counting kit 8 (CCK-8) and flow cytometry experiments were implemented to investigate the cell viability and apoptosis rate of MH7A cells. TargetScan was applied to identify the targets of miR-496, and the regulation of miR-496 on MMP10 expression was validated by a dual-luciferase reporter gene assay. qRT-PCR and western blot analyses were conducted to examine the expression. miR-496 expression was decreased in RA tissues and MH7A cells after IL-1β treatment. Overexpression of miR-496 significantly inhibited IL-1β-treated MH7A cell viability. MMP10 was identified as a target of miR-496 and its expression was negatively regulated by miR-496. The effects of miR-496 on MH7A cell proliferation and apoptosis were reversed by MMP10. The activity of NF-κB pathway was associated with the miR-496/MMP10 axis in IL-1β-stimulated MH7A cells. To summarize, this study demonstrated that miR-496 can impair the proliferative ability and facilitate the apoptosis of IL-1β-treated MH7A through regulating MMP10 expression and NF-κB signaling pathway.
类风湿关节炎（RA）是一种常见的慢性自身免疫性疾病，其特征为滑膜炎症。miR-496 密切参与各种病理状态。然而，其在 RA 中的作用尚未阐明。根据从基因表达综合数据库（GEO）数据集检索到的 RA 临床样本，确定 miR-496 和 MMP10 的表达。在由 IL-1β（10ng/mL）刺激的 MH7A 细胞中构建 RA 的体外模型。实施细胞计数试剂盒 8（CCK-8）和流式细胞术实验，以研究 MH7A 细胞的细胞活力和凋亡率。应用 TargetScan 鉴定 miR-496 的靶标，并通过双荧光素酶报告基因检测验证 miR-496 对 MMP10 表达的调控。进行 qRT-PCR 和 Western blot 分析以检查表达。miR-496 在 RA 组织和 IL-1β 处理后的 MH7A 细胞中表达降低。miR-496 的过表达显著抑制了 IL-1β 处理的 MH7A 细胞活力。MMP10 被鉴定为 miR-496 的靶标，其表达受 miR-496 的负调控。miR-496 对 MH7A 细胞增殖和凋亡的影响通过 MMP10 被逆转。NF-κB 通路的活性与 IL-1β 刺激的 MH7A 细胞中的 miR-496/MMP10 轴有关。总之，本研究表明，miR-496 通过调节 MMP10 表达和 NF-κB 信号通路，可损害 IL-1β 处理的 MH7A 的增殖能力并促进其凋亡。
