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Abstract：
BACKGROUND: This study aims to explore how miR-98-5p affects osteoarthritis, focusing on its role in chondrocyte inflammation, apoptosis, and extracellular matrix (ECM) degradation.
METHODS: Quantitative real-time PCR was used to measure miR-98-5p and CASP3 mRNA levels in OA cartilage tissues and IL-1β-treated CHON-001 cells. We predicted miR-98-5p and CASP3 binding sites using TargetScan and confirmed them via luciferase reporter assays. Chondrocyte viability was analyzed using CCK-8 assays, while pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) were quantified via ELISA. Caspase-3 activity was examined to assess apoptosis, and Western blotting was conducted for protein marker quantification.
RESULTS: Our results showed lower miR-98-5p levels in both OA cartilage and IL-1β-stimulated cells. Increasing miR-98-5p resulted in reduced pro-inflammatory cytokines, decreased caspase-3 activity, and improved cell viability. Furthermore, miR-98-5p overexpression hindered IL-1β-induced ECM degradation, evident from the decline in MMP-13 and β-catenin levels, and an increase in COL2A1 expression. MiR-98-5p\'s impact on CASP3 mRNA directly influenced its expression. Mimicking miR-98-5p\'s effects, CASP3 knockdown also inhibited IL-1β-induced inflammation, apoptosis, and ECM degradation. In contrast, CASP3 overexpression negated the suppressive effects of miR-98-5p.
CONCLUSIONS: In conclusion, our data collectively suggest that miR-98-5p plays a protective role against IL-1β-induced damage in chondrocytes by targeting CASP3, highlighting its potential as a therapeutic target for OA.
METHODS: Quantitative real-time PCR was used to measure miR-98-5p and CASP3 mRNA levels in OA cartilage tissues and IL-1β-treated CHON-001 cells. We predicted miR-98-5p and CASP3 binding sites using TargetScan and confirmed them via luciferase reporter assays. Chondrocyte viability was analyzed using CCK-8 assays, while pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) were quantified via ELISA. Caspase-3 activity was examined to assess apoptosis, and Western blotting was conducted for protein marker quantification.
RESULTS: Our results showed lower miR-98-5p levels in both OA cartilage and IL-1β-stimulated cells. Increasing miR-98-5p resulted in reduced pro-inflammatory cytokines, decreased caspase-3 activity, and improved cell viability. Furthermore, miR-98-5p overexpression hindered IL-1β-induced ECM degradation, evident from the decline in MMP-13 and β-catenin levels, and an increase in COL2A1 expression. MiR-98-5p\'s impact on CASP3 mRNA directly influenced its expression. Mimicking miR-98-5p\'s effects, CASP3 knockdown also inhibited IL-1β-induced inflammation, apoptosis, and ECM degradation. In contrast, CASP3 overexpression negated the suppressive effects of miR-98-5p.
CONCLUSIONS: In conclusion, our data collectively suggest that miR-98-5p plays a protective role against IL-1β-induced damage in chondrocytes by targeting CASP3, highlighting its potential as a therapeutic target for OA.
摘要：
背景：本研究旨在探讨miR-98-5p如何影响骨关节炎，专注于它在软骨细胞炎症中的作用，凋亡，和细胞外基质(ECM)降解。
方法：定量实时PCR检测OA软骨组织和IL-1β处理的CHON-001细胞中miR-98-5p和CASP3mRNA水平。我们使用TargetScan预测了miR-98-5p和CASP3结合位点，并通过荧光素酶报告基因测定证实了它们。使用CCK-8测定法分析软骨细胞活力，而促炎细胞因子(IL-1β，IL-6，TNF-α）通过ELISA定量。检测Caspase-3活性以评估细胞凋亡，和蛋白质印迹进行蛋白质标记定量。
结果：我们的结果显示，在OA软骨和IL-1β刺激的细胞中，miR-98-5p水平较低。增加miR-98-5p导致促炎细胞因子减少，caspase-3活性降低，和提高细胞活力。此外，miR-98-5p过表达阻碍IL-1β诱导的ECM降解，从MMP-13和β-catenin水平的下降可以明显看出，和COL2A1表达增加。MiR-98-5p对CASP3mRNA的影响直接影响其表达。模仿miR-98-5p的效果，CASP3敲低也抑制IL-1β诱导的炎症，凋亡，和ECM降解。相比之下,CASP3过表达否定了miR-98-5p的抑制作用。
结论：结论：我们的数据共同表明,miR-98-5p通过靶向CASP3对IL-1β诱导的软骨细胞损伤具有保护作用,突出了其作为OA治疗靶点的潜力.
方法：定量实时PCR检测OA软骨组织和IL-1β处理的CHON-001细胞中miR-98-5p和CASP3mRNA水平。我们使用TargetScan预测了miR-98-5p和CASP3结合位点，并通过荧光素酶报告基因测定证实了它们。使用CCK-8测定法分析软骨细胞活力，而促炎细胞因子(IL-1β，IL-6，TNF-α）通过ELISA定量。检测Caspase-3活性以评估细胞凋亡，和蛋白质印迹进行蛋白质标记定量。
结果：我们的结果显示，在OA软骨和IL-1β刺激的细胞中，miR-98-5p水平较低。增加miR-98-5p导致促炎细胞因子减少，caspase-3活性降低，和提高细胞活力。此外，miR-98-5p过表达阻碍IL-1β诱导的ECM降解，从MMP-13和β-catenin水平的下降可以明显看出，和COL2A1表达增加。MiR-98-5p对CASP3mRNA的影响直接影响其表达。模仿miR-98-5p的效果，CASP3敲低也抑制IL-1β诱导的炎症，凋亡，和ECM降解。相比之下,CASP3过表达否定了miR-98-5p的抑制作用。
结论：结论：我们的数据共同表明,miR-98-5p通过靶向CASP3对IL-1β诱导的软骨细胞损伤具有保护作用,突出了其作为OA治疗靶点的潜力.
