Immunology Division, Department of Microbiology, Faculty of Medicine, Chulalongkorn University, 1873 Rama 4 Road, Bangkok, 10330, Thailand.
Medical Microbiology Interdisciplinary Program, Graduate School, Chulalongkorn University, Bangkok, Thailand.
Sci Rep. 2024 Aug 5;14(1):18043. doi: 10.1038/s41598-024-68404-9.
In osteoarthritis (OA), extracellular matrix (ECM) digestion by cartilage-degrading enzymes drives cartilage destruction and generates ECM fragments, such as proteoglycan aggrecan (PG) peptides. PG peptides have been shown to induce immunological functions of chondrocytes. However, the role of PG peptides in stimulating catabolic mediators from chondrocytes has not been investigated. Therefore, we aim to determine the effects and its mechanism by which PG peptides induce chondrocytes to produce catabolic mediators in OA. Human chondrocytes were stimulated with IFNγ and various PG peptides either (i) with or (ii) without TLR2 blockade or (iii) with Lactobacillus species-conditioned medium (LCM), a genus of bacteria with anti-inflammatory properties. Transcriptomic analysis, cartilage-degrading enzyme production and TLR2-intracellular signaling activation were investigated. Chondrocytes treated with PG peptides p16-31 and p263-280 increased expression levels of genes associated with chondrocyte hypertrophy, cartilage degradation and proteolytic enzyme production. TLR2 downstream signaling proteins (STAT3, IkBα and MAPK9) were significantly phosphorylated in p263-280 peptide-stimulated chondrocytes. MMP-1 and ADAMTS-4 were significantly reduced in p263-280 peptides-treated condition with TLR2 blockade or LCM treatment. Phosphorylation levels of IkBa, ERK1/2 and MAPK9 were significantly decreased with TLR2 blockade, but only phosphorylation levels of MAPK9 was significantly decreased with LCM treatment. Our study showed that PG peptide stimulation via TLR2 induced cartilage-degrading enzyme production via activation of MAPK, NFκB and STAT3 pathways.
在骨关节炎(OA)中,软骨降解酶对细胞外基质(ECM)的消化作用导致软骨破坏,并产生 ECM 片段,如蛋白聚糖聚集素(PG)肽。已经表明 PG 肽可诱导软骨细胞的免疫功能。然而,PG 肽在刺激软骨细胞产生分解代谢介质方面的作用尚未得到研究。因此,我们旨在确定 PG 肽诱导 OA 中软骨细胞产生分解代谢介质的作用及其机制。用 IFNγ和各种 PG 肽刺激人软骨细胞,(i)有或(ii)无 TLR2 阻断或(iii)用具有抗炎特性的细菌属乳杆菌属条件培养基(LCM)。研究了转录组分析、软骨降解酶的产生和 TLR2 细胞内信号转导的激活。用 PG 肽 p16-31 和 p263-280 处理的软骨细胞增加了与软骨细胞肥大、软骨降解和蛋白水解酶产生相关的基因的表达水平。TLR2 下游信号蛋白(STAT3、IkBα 和 MAPK9)在 p263-280 肽刺激的软骨细胞中被显著磷酸化。在 TLR2 阻断或 LCM 处理的 p263-280 肽处理条件下,MMP-1 和 ADAMTS-4 显著减少。TLR2 阻断可显著降低 IkBa、ERK1/2 和 MAPK9 的磷酸化水平,但仅用 LCM 处理可显著降低 MAPK9 的磷酸化水平。我们的研究表明,PG 肽通过 TLR2 刺激通过激活 MAPK、NFκB 和 STAT3 通路诱导软骨降解酶的产生。