Libo Yuan, Ling Yao, Xianzhen Ren, Xusheng Chen, Kaiqiang Kang, Yongqing Xu, Tao Jin
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引用次数: 0
摘要
热休克是骨关节炎(OA)发病过程中的一种新型细胞死亡方式,但其潜在机制尚未完全明了。本研究旨在研究Runx2在软骨细胞(CH)热噬中的作用,并探讨其对Caspase-1表达的影响。研究人员收集了无 OA 病史的骨折患者的人体膝关节组织。用 siRNA 和 CADD522 阻断 Runx2 的功能。用 MTT 检测细胞活力。通过 RT-PCR、免疫荧光或 Western blot 检测 Runx2、Caspase1/4/5/11、GSDMD、胶原蛋白-II、凝集素、IL-1β、IL-8 和 MMP3/9 的表达水平。此外,DNA免疫沉淀和荧光素酶报告实验也证实了Runx2和Caspase-1之间的转录关联。Runx2 和 Caspase-1 在 LPS 处理的 CHs 中表达增加。Runx2与Caspase-1的启动子结合并激活其表达。此外,沉默Runx2或破坏Runx2的DNA结合能力可减轻LPS诱导的嗜热表型,包括Caspase-1激活、胶原蛋白II和凝集素降解、活力抑制、IL-1β和IL-8上调。阻断Runx2的表达或功能能以Caspase-1的方式缓解LPS引起的CHs热脓毒症,这表明人们对OA的病理有了新的认识。
Blocking Runx2 Inhibits the Caspase-1 Dependent Pyroptosis in Lipopolysaccharide-Treated Chondrocyte
Pyroptosis is a new type of cell death in the development of osteoarthritis (OA), but the underlying mechanism is not fully understood. This study aimed to investigate the role of Runx2 in the pyroptosis of chondrocyte (CH) and explore its effect on Caspase-1 expression. Human knee
tissues from the fracture patients without OA history were collected. Human CHs isolated from the tissue were treated by lipopolysaccharide (LPS) to establish the model of OA. siRNA and CADD522 were used to block the function of Runx2. The cell viability was tested by MTT. The expression levels
of Runx2, Caspase1/4/5/11, GSDMD, collagen-II, aggrecan, IL-1β, IL-8, and MMP3/9 were detected by RT-PCR, immunofluorescence, or western blot. Besides, the transcriptional association between Runx2 and Caspase-1 was confirmed by DNA immunoprecipitation and luciferase reporter assay.
Runx2 and Caspase-1 expression were increased in LPS-treated CHs. Runx2 bound to the promoter of Caspase-1 and activated its expression. Moreover, silencing Runx2 or disrupting the DNA-binding ability of Runx2 attenuated the LPS-induced pyroptotic phenotype, containing Caspase-1 activation,
collagen-II and aggrecan degradation, viability suppression, IL-1β and IL-8 upregulation. Blocking the expression or function of Runx2 alleviated the LPS-caused pyroptosis in CHs in the Caspase-1 manner, indicating a novel understanding of the pathology of OA.