ALOX5敲除通过抑制NF-κB信号通路减轻骨关节炎的软骨损伤
ALOX5 knockdown alleviates cartilage damage in osteoarthritis by inhibiting the NF-κB signaling pathway.
摘要(中文译文)
本研究旨在阐明花生四烯酸5-脂氧合酶（ALOX5）在骨关节炎（OA）中的作用及其潜在机制，以寻找该疾病的新治疗靶点。收集5例正常软骨组织和5例OA软骨组织样本，随后进行转录组测序以分析OA中的差异表达基因（DEGs）。采用免疫组织化学（IHC）、蛋白质印迹和GEO数据库分析验证ALOX5在人类和大鼠OA软骨中的表达。分别采用IL-1β和内侧半月板失稳（DMM）建立体外和体内OA模型，以评估ALOX5在铁蓄积、ROS生成、炎性细胞因子释放、铁死亡和细胞外基质（ECM）降解中的作用。此外，进行KEGG-GSEA以确定ALOX5调控的关键下游通路。进一步采用蛋白质印迹评估ALOX5对NF-κB通路的影响，并向软骨细胞给予NF-κB激活剂桦木酸（BA）以验证ALOX5在该通路中的调控作用。ALOX5在OA软骨组织和IL-1β诱导的软骨细胞中表达显著上调。敲低ALOX5可减弱促炎细胞因子（包括IL-6和TNF-α）的产生，减少铁死亡相关标志物如ROS和Fe的蓄积，并抑制关键ECM降解相关因子（包括MMP3、MMP13和ADAMTS5）的表达。相反，沉默ALOX5导致铁死亡抑制剂GPX4和SLC7A11显著上调，并恢复主要软骨成分II型胶原的表达。在OA大鼠中，关节腔内注射AAV-shALOX5可显著减轻软骨损伤。此外，敲低ALOX5可显著降低p-p65/p65和p-IKKβ/IKKβ的表达比值。值得注意的是，BA处理可逆转ALOX5敲低所赋予的保护作用，恢复NF-κB通路激活，继而促进软骨细胞铁死亡和ECM降解。ALOX5在OA中表达上调。敲低ALOX5通过失活NF-κB信号通路，从而抑制软骨细胞铁死亡和ECM降解，减轻OA进展。
Abstract
This study aims to elucidate the role and underlying mechanisms of arachidonate 5-lipoxygenase (ALOX5) in osteoarthritis (OA) to identify novel therapeutic targets for this disease. Five samples of normal cartilage tissue and five samples of OA cartilage tissue were collected, followed by transcriptome sequencing was performed to analyze differentially expressed genes (DEGs) in OA. Immunohistochemistry (IHC), western blot, and analysis of the GEO database were used to verify ALOX5 expression in both humans and rat OA cartilage. IL-1β and destabilization of the medial meniscus (DMM) were employed to establish invitro and in vivo OA models, respectively, to assess the role of ALOX5 in Fe accumulation, ROS generation, inflammatory cytokines release, ferroptosis, and extracellular matrix (ECM) degradation. Additionally, KEGG-GSEA was conducted to identify key downstream pathways regulated by ALOX5. Furthermore, western blot was used to assess the effect of ALOX5 on the NF-κB pathway, and the NF-κB activator betulinic acid (BA) was administered to chondrocytes to validate the regulatory role of ALOX5 in this pathway. ALOX5 expression was significantly upregulated in OA cartilage tissues and in IL-1β-induced chondrocytes. Knockdown of ALOX5 attenuated the production of pro-inflammatory cytokines, including IL-6 and TNF-α; reduced the acculation of ferroptosis-associated markers such as ROS and Fe; and suppressed the ezpression of key ECM degradation-related factors, including MMP3, MMP13, and ADAMTS5. Conversely, ALOX5 silencing led to a significant upregulation of ferroptosis inhibitors GPX4 and SLC7A11, as well as trstored expression of collagen II, the major cartilage component. In OA rats, intra-articular injection of AAV-shALOX5 significantly alleviated cartilage damage. Moreover, ALOX5 knockdown significantly reduced the expression ratios of p-p65/p65 and p-IKKβ/IKKβ. Notably, treatment with BA reversed the protective effects conferred by ALOX5 knockdown, restoring NF-κB pathway activation and subsequently promoting ferroptosis and ECM degradation in chondrocytes. ALOX5 is upregulated in OA. Knockdown of ALOX5 alleviates OA progression by inactivating the NF-κB signaling pathway, thereby inhibiting ferroptosis and ECM degradation in chondrocytes.
如何引用
AMA
Gang Cheng, Kaiwen Chen, Zhigang Wang, Huihai Chen, Yixin Shen. ALOX5 knockdown alleviates cartilage damage in osteoarthritis by inhibiting the NF-κB signaling pathway.. Cytokine. 2026; doi:10.1016/j.cyto.2026.157150.
APA
Gang Cheng, Kaiwen Chen, Zhigang Wang, Huihai Chen, Yixin Shen (2026). ALOX5 knockdown alleviates cartilage damage in osteoarthritis by inhibiting the NF-κB signaling pathway.. Cytokine. https://doi.org/10.1016/j.cyto.2026.157150
GB/T 7714
Gang Cheng, Kaiwen Chen, Zhigang Wang, Huihai Chen, Yixin Shen. ALOX5 knockdown alleviates cartilage damage in osteoarthritis by inhibiting the NF-κB signaling pathway.[J]. Cytokine, 2026 doi:10.1016/j.cyto.2026.157150.
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