Department of Rheumatology, Gansu Provincial Hospital of Traditional Chinese Medicine, No. 418, Guazhou Road, Qilihe District, Lanzhou City, 730050, Gansu Province, China.
Department of Acupuncture and Moxibustion, Gansu Provincial Hospital of Traditional Chinese Medicine, No. 418, Guazhou Road, Qilihe District, Lanzhou City, 730050, Gansu Province, China.
Inflammation. 2020 Dec;43(6):2048-2060. doi: 10.1007/s10753-019-01118-7. Epub 2020 Sep 29.
Studies have found that N-myc downstream-regulated gene 2 (Ndrg2) is involved in the progression of rheumatoid arthritis (RA); however, the specific mechanism still remains unclear. Gene expression profiles in the tibial joints of the collagen-induced rheumatoid arthritis model were obtained using Gene Expression Omnibus database. Western blot and real-time PCR were respectively performed to determine the expression of Ndrg2 and gene messenger RNA. Cell viability was measured by Cell Counting Kit-8 (CCK-8) method, and cell cycle was detected by flow cytometry. Cell scratch assays were carried out to detect migration. The binding ability of miR-130a to Ndrg2-3'-UTR was predicted by TargetScan website and confirmed by dual luciferase assay. A collagen-induced arthritis rat model was constructed to observe the effects of miR-130a on arthritis index, hind limb swelling, volume of rat hind paw, and inflammation. Ndrg2 was found downregulated in RA tissues, and knockdown of Ndrg2 promoted fibroblast-like synoviocytes (FLS) proliferation and inflammation, while overexpressed Ndrg2 produced opposite results. Ndrg2 was predicted as a target gene for miR-130a, and miR-130a mimic promoted FLS proliferation, while miR-130a inhibitor suppressed FLS proliferation. Moreover, we found that miR-130a antagomir could significantly reduce the arthritis index, swelling degree, foot volume, and inflammatory factor levels; inhibit the expression of miR-130a; and promote the expression of Ndrg2. The miR-130a/Ndrg2 axis signaling pathway is involved in the progression of RA. Our findings provide a theoretical basis for the clinical treatment of RA.
研究发现 N- 小细胞肺癌下游调节基因 2（Ndrg2）参与类风湿关节炎（RA）的进展；然而，具体机制仍不清楚。使用基因表达综合数据库获得胶原诱导性类风湿关节炎模型胫骨关节的基因表达谱。分别通过 Western blot 和实时 PCR 测定 Ndrg2 和基因信使 RNA 的表达。通过细胞计数试剂盒 -8（CCK-8）法测定细胞活力，通过流式细胞术检测细胞周期。进行细胞划痕实验检测迁移。TargetScan 网站预测 miR-130a 与 Ndrg2-3'-UTR 的结合能力，并通过双荧光素酶测定验证。构建胶原诱导性关节炎大鼠模型，观察 miR-130a 对关节炎指数、后肢肿胀、大鼠后爪体积和炎症的影响。发现 Ndrg2 在 RA 组织中下调，下调 Ndrg2 促进成纤维样滑膜细胞（FLS）增殖和炎症，而过表达 Ndrg2 则产生相反的结果。Ndrg2 被预测为 miR-130a 的靶基因，miR-130a 模拟物促进 FLS 增殖，而 miR-130a 抑制剂抑制 FLS 增殖。此外，我们发现 miR-130a 拮抗剂可显著降低关节炎指数、肿胀程度、足体积和炎症因子水平；抑制 miR-130a 的表达；并促进 Ndrg2 的表达。miR-130a/Ndrg2 轴信号通路参与 RA 的进展。我们的研究结果为 RA 的临床治疗提供了理论依据。
