Department of Health Management, Changzhi Medical College, Changzhi, Shanxi Province, China.
Department of Sports Medicine, Military Sports Training Center of the Training Management Department of the Central Military Commission, Beijing, China;
Allergol Immunopathol (Madr). 2022 Nov 1;50(6):169-175. doi: 10.15586/aei.v50i6.744. eCollection 2022.
To reveal the possible effects of death-associated protein kinase 1 (DAPK1) on the progression of osteoarthritis (OA) and the potential underlying mechanism.
The expression of DAPK1 in OA and normal samples and interleukin (IL)-1β-stimulated chondrocytes was analyzed by quantitative real-time polymerase chain reaction and Immunoblot assay. Cell viability, proliferation, and apoptosis in DAPK1-knockdown cells stimulated with IL-1β were detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution, 5-ethynyl-2β-deoxyuridine staining and flow cytometry. The chondrocyte degradation and inflammatory response in IL-1β-induced chondrocytes were investigated by Immunoblot analysis and enzyme-linked-immunosorbent serologic assay. In addition, the effect of DAPK1 on p38 mitogen-activated protein kinase (MAPK) activation was analyzed by immunoblot assay.
This study revealed that DAPK1 was highly expressed in OA patients and IL-1β-induced chondrocytes. Down-regulation of DAPK1 enhanced IL-1β-induced chondrocyte proliferation. DAPK1 knockdown inhibited IL-1β-induced chondrocyte degradation. In addition, DAPK1 depletion inhibited IL-1β-induced chondrocyte inflammation. Mechanically, it was revealed that down--regulation of DAPK1 could inhibit the p38 MAPK pathway, and therefore affected progression of OA.
DAPK1 knockdown attenuates IL-1β-induced extracellular matrix degradation and inflammatory response in OA chondrocytes by regulating the p38 MAPK pathway.
揭示死亡相关蛋白激酶 1（DAPK1）对骨关节炎（OA）进展的可能影响及其潜在机制。
通过定量实时聚合酶链反应和免疫印迹法分析 OA 和正常样本以及白细胞介素（IL）-1β刺激的软骨细胞中 DAPK1 的表达。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）溶液、5-乙炔基-2β-脱氧尿苷染色和流式细胞术检测 DAPK1 敲低细胞在 IL-1β刺激下的细胞活力、增殖和凋亡。通过免疫印迹分析和酶联免疫吸附血清测定法研究 IL-1β诱导的软骨细胞中的软骨细胞降解和炎症反应。此外，通过免疫印迹法分析 DAPK1 对 p38 丝裂原活化蛋白激酶（MAPK）激活的影响。
本研究表明 DAPK1 在 OA 患者和 IL-1β诱导的软骨细胞中高表达。下调 DAPK1 增强了 IL-1β诱导的软骨细胞增殖。DAPK1 敲低抑制了 IL-1β诱导的软骨细胞降解。此外，DAPK1 耗竭抑制了 IL-1β诱导的软骨细胞炎症。在机制上，下调 DAPK1 可以抑制 p38 MAPK 通路，从而影响 OA 的进展。
通过调节 p38 MAPK 通路，DAPK1 敲低可减轻 IL-1β诱导的 OA 软骨细胞细胞外基质降解和炎症反应。
