Department of Geriatrics, Tangshan Gongren Hospital, Tangshan, Hebei, China.
Autoimmunity. 2022 Sep;55(6):371-377. doi: 10.1080/08916934.2022.2081842. Epub 2022 Jun 6.
Fibroblast-like synoviocytes (FLS) are crucial regulators in the pathogenesis of rheumatoid arthritis (RA). Reportedly, microRNA (miR) participates in regulating the pathogenesis of RA. In this study, we explored the regulatory effects of miR-300 on the proliferation, migration and invasion of FLS, which were obtained from RA patients.
qPCR was utilized to detect miR-300 expression and interleukin-37 (IL-37) mRNA expression in the synovial tissue of RA patients and healthy controls. Cell counting kit-8 (CCK-8) assay and Transwell assay were performed to investigate the regulatory function of miR-300 on the proliferation, migration and invasion of FLS. ELISA was employed to detect TNF-α, IL-6 and IL-8 levels, to evaluate the inflammatory response. Bioinformatics analysis and luciferase reporter assay were applied to validate the targeting relationship between miR-300 and IL-37. Western blot assay was executed to detect IL-37 protein expression in FLS.
MiR-300 was revealed to be markedly down-modulated in the synovial tissue and FLS of RA patients; meanwhile, IL-37 expression was up-modulated. The transfection of miR-300 mimics enhanced RA-FLS growth, migration, invasion and inflammatory response; transfection of miR-300 inhibitors repressed the growth, migration, invasion and inflammatory response of RA-FLS. IL-37 was identified as a downstream target of miR-300, and IL-37 partially counteracted the enhanced growth, migration, invasion and inflammatory response of RA-FLS induced by miR-300.
MiR-300 facilitates growth, migration, invasion and inflammatory response of FLS by targeting IL-37, suggesting it was a crucial regulator in the pathogenesis of RA.
成纤维样滑膜细胞（FLS）是类风湿关节炎（RA）发病机制中的关键调节细胞。据报道，微小 RNA（miRNA）参与调控 RA 的发病机制。在这项研究中，我们探讨了 miR-300 对 RA 患者来源的 FLS 增殖、迁移和侵袭的调节作用。
qPCR 检测 RA 患者和健康对照者滑膜组织中 miR-300 表达和白细胞介素 37（IL-37）mRNA 表达。细胞计数试剂盒-8（CCK-8）检测和 Transwell 检测 miR-300 对 FLS 增殖、迁移和侵袭的调节作用。酶联免疫吸附试验（ELISA）检测肿瘤坏死因子-α（TNF-α）、白细胞介素 6（IL-6）和白细胞介素 8（IL-8）水平，评估炎症反应。生物信息学分析和荧光素酶报告基因检测验证 miR-300 与 IL-37 的靶向关系。Western blot 检测 FLS 中 IL-37 蛋白表达。
miR-300 在 RA 患者滑膜组织和 FLS 中表达明显下调，同时 IL-37 表达上调。miR-300 模拟物转染增强 RA-FLS 生长、迁移、侵袭和炎症反应；miR-300 抑制剂转染抑制 RA-FLS 生长、迁移、侵袭和炎症反应。IL-37 被鉴定为 miR-300 的下游靶点，IL-37 部分抵消了 miR-300 诱导的 RA-FLS 生长、迁移、侵袭和炎症反应增强。
miR-300 通过靶向 IL-37 促进 FLS 的生长、迁移、侵袭和炎症反应，提示其在 RA 发病机制中是一个关键调节因子。
