Farrag Mariam, Cordero-Barreal Alfonso, Ait Eldjoudi Djedjiga, Varela-García María, Torrijos Pulpón Carlos, Lago Francisca, Essawy Amina, Soffar Ahmed, Pino Jesus, Farrag Yousof, Gualillo Oreste
Research Laboratory 9, NEIRID Lab Neuroendocrine Interactions in Rheumatology and Inflammatory Diseases, SERGAS Servizo Galego de Saude, IDIS Instituto de Investigación Sanitaria de Santiago, Santiago University Clinical Hospital, Santiago de Compostela, Spain.
Euro-Mediterranean Master in Neuroscience and Biotechnology, Faculty of Science, Alexandria University, Alexandria, Egypt.
Cartilage. 2024 Oct 29:19476035241292323. doi: 10.1177/19476035241292323.
This study investigates how the inflammatory response of ATDC5 murine chondrogenic cells influences the activity of C6 (rat) and GL261 (mouse) glial cell lines. Prior research suggested nitric oxide (NO) involvement in cartilage-immune crosstalk. The current study explores whether NO, produced by inflamed chondrocytes, mediates signaling between chondrocytes and glial cells.
Pre-challenged ATDC5 cells with 250 ng/ml of lipopolysaccharide (LPS) were cocultured with GL261 or C6 glioma cells for 24 h with a transwell culture system. Cell viability was assessed using MTT assay. Gene and protein expression were evaluated by qRT-PCR and WB, respectively.
Real-time reverse transcription-polymerase chain reaction (RT-qPCR) indicated statistically significant upregulation of LCN2, IL-6, TNF-α, IL-1β, and GFAP in glial cells following 24-h coculture with challenged ATDC5 cells. Suppression of LPS-induced NO production by aminoguanidine decreased LPS-mediated LCN2 and IL-6 expression in glioma cells. We identified also the involvement of the ERK1/2 and AKT signaling pathways in the glial neuroinflammatory response.
This study demonstrates, for the first time, that NO produced by inflamed murine chondrocytes mediated pro-inflammatory responses in glial cells via ERK1/2 and AKT signaling, highlighting a potential mechanism linking cartilage NO to neuroinflammation and chronic pain in osteoarthritis.
本研究调查了ATDC5小鼠软骨生成细胞的炎症反应如何影响C6（大鼠）和GL261（小鼠）神经胶质瘤细胞系的活性。先前的研究表明一氧化氮（NO）参与软骨与免疫的相互作用。本研究探讨了炎症软骨细胞产生的NO是否介导软骨细胞与神经胶质瘤细胞之间的信号传导。
将预先用250 ng/ml脂多糖（LPS）刺激的ATDC5细胞与GL261或C6神经胶质瘤细胞使用Transwell培养系统共培养24小时。使用MTT法评估细胞活力。分别通过qRT-PCR和WB评估基因和蛋白质表达。
实时逆转录聚合酶链反应（RT-qPCR）表明，与受刺激的ATDC5细胞共培养24小时后，神经胶质瘤细胞中LCN2、IL-6、TNF-α、IL-1β和GFAP的表达在统计学上有显著上调。氨基胍抑制LPS诱导的NO产生可降低LPS介导的神经胶质瘤细胞中LCN2和IL-6的表达。我们还确定了ERK1/2和AKT信号通路参与了神经胶质瘤细胞的神经炎症反应。
本研究首次证明，炎症小鼠软骨细胞产生的NO通过ERK1/2和AKT信号传导介导神经胶质瘤细胞中的促炎反应，突出了软骨NO与骨关节炎神经炎症和慢性疼痛之间潜在联系的机制。
