Department of Orthopaedics, Luoyang Orthopedic Hospital of Henan Province (Orthopedic Hospital of Henan Province), No. 82 South Qi Ming Road, Luoyang, 471002, Henan, China.
Inflamm Res. 2020 Nov;69(11):1123-1132. doi: 10.1007/s00011-020-01392-4. Epub 2020 Aug 18.
Osteoarthritis (OA) impacts the quality of life in middle-aged and elderly people by inducing immobility. The severe inflammation in chondrocytes is reported to be related to the development and process of OA. The present study aims to investigate the protective effects of Apremilast on injured chondrocytes induced by interleukin-1α (IL-1α) and the underlying mechanism.
10 ng/mL IL-1α was used to induce the in vitro injured chondrocytes. QRT-PCR was used to evaluate the expression level of Sry-type high-mobility-group box 9 (SOX-9), collagen type II alpha-1 gene (COL2A1), Aggrecan (ACAN) and collagen type X alpha 1 chain (COL10A1). SiRNA technology was utilized to knock down the expression of SOX-9 in the chondrocytes. The expression of SOX-9 was determined by Western Blot assay and/or immunofluorescence assay. Western Blot was used to evaluate the expression level of phosphorylated cyclic AMP response element binding (CREB).
SOX9, Col2a1 and Acan were significantly up-regulated and Col10a1 was significantly down-regulated in the chondrocytes by Apremilast in a dose-dependent manner. IL-1α induced the injured chondrocytes by decreasing the expression of SOX9, Col2a1, Acan and increasing the expression of Col10a1, which were greatly reversed by Apremilast. By silencing SOX-9, the effects of Apremilast on SOX9 and marker genes were abolished. Phosphorylated CREB was up-regulated by Apremilast in a time-dependent manner. The up-regulated SOX-9 by Apremilast was reversed by the protein kinase A (PKA)/CREB pathway inhibitor H89.
Apremilast may protect chondrocytes from inflammation by up-regulating SOX9.
骨关节炎（OA）通过导致活动受限而影响中老年人的生活质量。据报道，软骨细胞中的严重炎症与 OA 的发展和进程有关。本研究旨在探讨阿普米司特对白细胞介素-1α（IL-1α）诱导的受损软骨细胞的保护作用及其潜在机制。
采用 10ng/mlIL-1α诱导体外损伤的软骨细胞。采用 QRT-PCR 检测 Sry 型高迁移率族盒 9（SOX-9）、Ⅱ型胶原α-1 基因（COL2A1）、聚集蛋白聚糖（ACAN）和 X 型胶原α-1 链（COL10A1）的表达水平。利用 siRNA 技术敲低软骨细胞中 SOX-9 的表达。通过 Western Blot 检测和/或免疫荧光检测确定 SOX-9 的表达。Western Blot 用于评估磷酸化环磷腺苷反应元件结合蛋白（CREB）的表达水平。
阿普米司特呈剂量依赖性地上调软骨细胞中 SOX9、Col2a1 和 Acan 的表达，下调 Col10a1 的表达。IL-1α 通过降低 SOX9、Col2a1、Acan 的表达和增加 Col10a1 的表达诱导损伤的软骨细胞，而阿普米司特则能显著逆转这些作用。沉默 SOX-9 后，阿普米司特对 SOX9 和标记基因的作用被消除。阿普米司特呈时间依赖性地上调磷酸化 CREB。阿普米司特上调的 SOX-9 被蛋白激酶 A（PKA）/CREB 通路抑制剂 H89 逆转。
阿普米司特可能通过上调 SOX9 来保护软骨细胞免受炎症的侵害。
