Department of Rheumatology and Immunology, The First Affiliated Hospital of China Medical University, 155 Nanjing North Street, Heping District, Shenyang 110001, PR China.
Department of Sports Medicine and Joint Surgery, The First Affiliated Hospital of China Medical University, 155 Nanjing North Street, Heping District, Shenyang 110001, PR China.
Mol Immunol. 2022 Jul;147:71-80. doi: 10.1016/j.molimm.2022.04.015. Epub 2022 Apr 30.
Angiogenesis associates with chondrocytes differentiation in inflammatory arthritis. Interleukin (IL)- 1β stimulated SW1353 cells have a phenotype similar to this kind of chondrocytes. IL-17A, a target in T helper 17 (Th17)/IL-17 signaling pathways, was expressed by SW1353 cells. The study aimed to explore the role of IL-35 on angiogenesis in IL-1β stimulated SW1353 cells and its related signaling pathways.
Microarray dataset was downloaded from the Gene Expression Omnibus database of arthritis cartilage. The protein-protein interaction (PPI) was analyzed for IL-35, pro-angiogenic factors and the differentially expressed genes (DEGs). We studied the effects of IL-35 on proliferation and apoptosis in IL-1β stimulated SW1353 cells using cell counting kit-8 (CCK-8) assay and flow cytometry. The expression of pro-angiogenic factors and IL-17A were assessed by western blot and real-time PCR. Added plumbagin (inhibitor of IL-17A) to repeat the above experiment. The secretion of IL-17A was assessed by ELISA.
IL-35, pro-angiogenic factors interacted with DEGs to affect the function of arthritis chondrocytes. IL-35 promoted IL-1β-stimulated SW1353 cells proliferation, inhibited apoptosis, and decreased pro-angiogenic molecules and IL-17A expression in a concentration dependent manner. IL-35 inhibited IL-17A secretion in the supernatants of these cells. Blocking the Th17/IL-17 related pathways with plumbagin abolished the effects of IL-35 on IL-1β-stimulated SW1353 cells.
These results suggested that IL-35 regulated differentiation and pro-angiogenic molecules expression in IL-1β stimulated SW1353 cells via Th17/IL-17 related signaling pathways. Our findings may reveal the mechanisms of novel angiogenesis molecules in inflammatory chondrocyte lesion.
血管生成与炎症性关节炎中的软骨细胞分化有关。白细胞介素（IL）-1β刺激的 SW1353 细胞具有类似于这种软骨细胞的表型。白细胞介素 17A（IL-17A）是 Th17/IL-17 信号通路的靶点，在 SW1353 细胞中表达。本研究旨在探讨 IL-35 在 IL-1β刺激的 SW1353 细胞中的血管生成作用及其相关信号通路。
从关节炎软骨基因表达综合数据库下载微阵列数据集。分析 IL-35、促血管生成因子和差异表达基因（DEGs）的蛋白质-蛋白质相互作用（PPI）。我们使用细胞计数试剂盒-8（CCK-8）测定法和流式细胞术研究了 IL-35 对 IL-1β刺激的 SW1353 细胞增殖和凋亡的影响。通过 Western blot 和实时 PCR 评估促血管生成因子和 IL-17A 的表达。加入白花丹醌（IL-17A 的抑制剂）重复上述实验。通过 ELISA 评估 IL-17A 的分泌。
IL-35、促血管生成因子与 DEGs 相互作用，影响关节炎软骨细胞的功能。IL-35 以浓度依赖的方式促进 IL-1β 刺激的 SW1353 细胞增殖，抑制凋亡，并降低促血管生成分子和 IL-17A 的表达。IL-35 抑制这些细胞上清液中 IL-17A 的分泌。用白花丹醌阻断 Th17/IL-17 相关通路可消除 IL-35 对 IL-1β 刺激的 SW1353 细胞的影响。
这些结果表明，IL-35 通过 Th17/IL-17 相关信号通路调节 IL-1β 刺激的 SW1353 细胞的分化和促血管生成分子的表达。我们的发现可能揭示了炎症性软骨细胞病变中新的血管生成分子的机制。
