Department of Rheumatology, Shenzhen Futian Hospital for Rheumatic Diseases, Shenzhen, China.
J Interferon Cytokine Res. 2022 Jun;42(6):279-289. doi: 10.1089/jir.2021.0228.
Rheumatoid arthritis (RA) is a systemic immune disease. Rho family GTPase 3 (RND3) has been reported to play an important role in inflammatory diseases. In this study, the expression of RND3 in RA was analyzed by gene chips. After RND3 was overexpressed, cell counting kit-8 assay was to detect the viability of fibroblast-like synovial cells (RA-FLSs). Transwell assays were to appraise the migratory and invasive capacities of RA-FLSs. Enzyme-linked immunosorbent assay (ELISA) and Western blot analysis were to estimate inflammatory response. In addition, MMP3 and MMP9 levels were also tested by ELISA analysis. After forkhead box M1 (FOXM1) was overexpressed, RND3 expression was detected by Western blot. The transcriptional relationship between FOXM1 and RND3 was predicted by HumanTFDB and JASPAR databases. Luciferase reporter and chromatin immunoprecipitation assays verified the binding ability of FOXM1 and RND3. The role of FOXM1/RND3 axis in RA was detected again by functional experiments. Western blot detected the expression of Rho/ROCK pathway-related proteins. RND3 expression was downregulated in RA. Overexpression of RND3 reduced the proliferation, migration, invasion, and inflammation of RA-FLSs. RND3 was inhibited by FOXM1 transcription, and upregulated FOXM1 reduced the inhibitory effect of RND3 overexpression on cell growth and inflammation, which might be associated with the Rho/ROCK pathway. RND3 transcriptionally regulated by FOXM1 inhibited the migration and inflammation of RA-FLSs in RA through the Rho/ROCK pathway.
类风湿关节炎（RA）是一种系统性自身免疫性疾病。已有研究报道 Rho 家族 GTP 酶 3（RND3）在炎症性疾病中发挥重要作用。本研究通过基因芯片分析 RA 中 RND3 的表达。过表达 RND3 后，通过细胞计数试剂盒-8 检测成纤维样滑膜细胞（RA-FLSs）的活力。Transwell 检测评估 RA-FLSs 的迁移和侵袭能力。酶联免疫吸附测定（ELISA）和 Western blot 分析评估炎症反应。此外，还通过 ELISA 分析检测 MMP3 和 MMP9 水平。过表达叉头框 M1（FOXM1）后，通过 Western blot 检测 RND3 的表达。通过 HumanTFDB 和 JASPAR 数据库预测 FOXM1 和 RND3 之间的转录关系。荧光素酶报告基因和染色质免疫沉淀实验验证 FOXM1 和 RND3 之间的结合能力。通过功能实验再次检测 FOXM1/RND3 轴在 RA 中的作用。Western blot 检测 Rho/ROCK 通路相关蛋白的表达。在 RA 中 RND3 的表达下调。过表达 RND3 降低了 RA-FLSs 的增殖、迁移、侵袭和炎症。FOXM1 转录抑制 RND3，上调 FOXM1 降低了 RND3 过表达对细胞生长和炎症的抑制作用，这可能与 Rho/ROCK 通路有关。RND3 受 FOXM1 转录调控，通过 Rho/ROCK 通路抑制 RA-FLSs 的迁移和炎症，从而参与 RA 的发生发展。
