Department of Orthopedic Surgery, Liaocheng People's Hospital, Liaocheng, Shandong, China (mainland).
Department of Imaging, Liaocheng Infectious Disease Hospital, Liaocheng, Shandong, China (mainland).
Med Sci Monit. 2021 Dec 8;27:e930053. doi: 10.12659/MSM.930053.
BACKGROUND Chondrocytes play a vital role in the later stages of osteoarthritis (OA). The roles of chemokine (C-C motif) ligand 2 (CCL2) and its receptor, chemokine receptor 2 (CCR2), are as yet poorly elucidated in chondrocyte hypertrophy (CH). Here, we aimed to regulate the CCL2/CCR2 axis and explore its effect on progression of CH. MATERIAL AND METHODS Chondrocytes isolated from patients with OA were used in the present study. In vitro experiments were conducted to test hypertrophic gene and CCL2/CCR2 expression in chondrocyte degeneration caused by interleukin (IL)-17A or CCL2 protein stimulation. In addition, inhibition of CCL2 and CCR2 was used to assess the role of CCL2 and CCR2 blockade in CH. Relative gene expression was determined with real-time polymerase chain reaction, western blot, or immunofluorescence. Hypertrophic changes were assessed with cell area measurement. Moreover, the viability of chondrocytes was analyzed using an MTT assay and flow cytometry was used to assess cell apoptosis. RESULTS CCL2 and CCR2 were upregulated in IL-17A-treated chondrocytes. The exogenic CCL2 stimulation also promoted CH and increased the expression of Type 10 collagen, RUNX2, and IHH, which could be reversed via suppression of CCR2. Inhibition of CCL2 and CCR2 expression was sufficient to: 1) protect Type 2 collagen synthesis; 2) alleviate IL-17A-induced overexpression of Type 10 collagen, RUNX2, and IHH; and 3) improve chondrocyte proliferation and apoptosis. CONCLUSIONS Blockading the CCL2/CCR2 axis plays a role in delaying the development of CH.
软骨细胞在骨关节炎(OA)的后期阶段发挥着至关重要的作用。趋化因子(C-C 基序)配体 2(CCL2)及其受体趋化因子受体 2(CCR2)在软骨细胞肥大(CH)中的作用尚不清楚。在这里,我们旨在调节 CCL2/CCR2 轴,并探讨其对 CH 进展的影响。
本研究使用了来自 OA 患者的分离软骨细胞。进行了体外实验以测试白细胞介素(IL)-17A 或 CCL2 蛋白刺激引起的软骨细胞退变中的肥大基因和 CCL2/CCR2 表达。此外,抑制 CCL2 和 CCR2 用于评估 CCL2 和 CCR2 阻断在 CH 中的作用。使用实时聚合酶链反应、western blot 或免疫荧光法测定相对基因表达。通过细胞面积测量评估肥大变化。此外,通过 MTT 测定分析软骨细胞的活力,并使用流式细胞术评估细胞凋亡。
CCL2 和 CCR2 在 IL-17A 处理的软骨细胞中上调。外源性 CCL2 刺激也促进了 CH,并增加了 Type 10 胶原、RUNX2 和 IHH 的表达,这些表达可以通过抑制 CCR2 来逆转。抑制 CCL2 和 CCR2 的表达足以:1)保护 Type 2 胶原的合成;2)减轻 IL-17A 诱导的 Type 10 胶原、RUNX2 和 IHH 的过度表达;3)改善软骨细胞的增殖和凋亡。
阻断 CCL2/CCR2 轴在延缓 CH 发展中起作用。