Hu Yu-Chen, Huang Tzu-Ching, Hsieh Bau-Shan, Huang Li-Wen, Lin Jin-Seng, Hsu Han-Yin, Lee Chia-Chia, Chang Kee-Lung
Department of Biochemistry, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807378, Taiwan.
Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807378, Taiwan.
Nutrients. 2024 Dec 23;16(24):4417. doi: 10.3390/nu16244417.
Osteoarthritis (OA) is a chronic condition characterized by joint pain and disability, driven by excessive oxidative stress and inflammatory cytokine production in chondrocytes, resulting in cell death and cartilage matrix breakdown. Our previous study showed that in monosodium iodoacetate (MIA)-induced OA rats, oral administration of heat-killed subsp. 557 (LDL557) could significantly decrease OA progression.
Accordingly, we designed an in vitro cell culture study aimed at investigating the effects of heat-killed LDL557 extracts on chondrocytes using SW1353 cells (a human chondrosarcoma cell line) challenged with 5 μM MIA to mimic OA conditions.
The results showed that the 10 μg/mL LDL557 extracts protected SW1353 cells from MIA-induced death and reduced extracellular matrix (ECM) loss, as evaluated by toluidine blue O staining and extracellular matrix component synthesis with RT-qPCR measurement. This was achieved by decreasing the expression of MIA-induced pro-inflammatory cytokines, including IL-1β, IL-6, and TNF-α, while slightly increasing the MIA-suppressed expression of the anti-inflammatory cytokine IL-10, which were evidenced by RT-qPCR analysis. Moreover, the RT-qPCR evaluation also indicated that the LDL557 extracts slightly reduced the expression of COX-2 compared with the control, while it did not reduce the MIA-increased expression of microsomal prostaglandin E synthase-1 (mPGES-1). In addition, the LDL557 extracts influenced neither the matrix-degrading protease expressions measured via RT-qPCR nor the oxidative stress measured via fluorescence flow cytometry in the cells with or without the MIA challenge.
This study demonstrates that LDL557 extracts may protect chondrocytes from OA damage by reducing inflammation-related factors and thus mitigating cartilage matrix loss, suggesting LDL557 extracts are attractive alternatives for OA applications.
骨关节炎（OA）是一种以关节疼痛和功能障碍为特征的慢性疾病，由软骨细胞中过度的氧化应激和炎性细胞因子产生所驱动，导致细胞死亡和软骨基质分解。我们之前的研究表明，在碘乙酸钠（MIA）诱导的OA大鼠中，口服热灭活的 亚种557（LDL557）可显著减缓OA进展。
因此，我们设计了一项体外细胞培养研究，旨在使用SW1353细胞（一种人软骨肉瘤细胞系），通过用5μM MIA刺激来模拟OA条件，研究热灭活的LDL557提取物对软骨细胞的影响。
结果表明，10μg/mL的LDL557提取物可保护SW1353细胞免受MIA诱导的死亡，并减少细胞外基质（ECM）损失，这通过甲苯胺蓝O染色和RT-qPCR测量细胞外基质成分合成来评估。这是通过降低MIA诱导的促炎细胞因子（包括IL-1β、IL-6和TNF-α）的表达来实现的，同时略微增加MIA抑制的抗炎细胞因子IL-10的表达，RT-qPCR分析证明了这一点。此外，RT-qPCR评估还表明，与对照组相比，LDL557提取物略微降低了COX-2的表达，而它并未降低MIA增加的微粒体前列腺素E合酶-1（mPGES-1）的表达。此外，LDL557提取物对通过RT-qPCR测量的基质降解蛋白酶表达以及通过荧光流式细胞术测量的有无MIA刺激的细胞中的氧化应激均无影响。
本研究表明，LDL557提取物可能通过减少炎症相关因子从而减轻软骨基质损失来保护软骨细胞免受OA损伤，提示LDL557提取物是OA应用中具有吸引力的替代物。
