Department of Osteoarthrosis, Luoyang Orthopedic‑Traumatological Hospital of Henan Province (Henan Provincial Orthopedic Hospital), Zhengzhou, Henan 450000, P.R. China.
Health Management Center, Luoyang Orthopedic‑Traumatological Hospital of Henan Province (Henan Provincial Orthopedic Hospital), Zhengzhou, Henan 450000, P.R. China.
Mol Med Rep. 2021 Jun;23(6). doi: 10.3892/mmr.2021.12063. Epub 2021 Apr 21.
Chondrocyte apoptosis is an important factor in the development and progression of osteoarthritis (OA). Cryptotanshinone (CTS) can inhibit chondrocyte apoptosis, but the specific mechanism remains unknown. The aim of the present study was to explore how CTS may affect chondrocyte apoptosis. Reverse transcription‑quantitative PCR and western blotting were used to validate microRNA (miR)‑574‑5p, YY1‑associated factor 2 (YAF2), Bcl‑2 and Bax expression levels. H&E, Safranin O and TUNEL staining assays were used to evaluate the apoptosis of arthritic chondrocytes . A Cell Counting Kit‑8 assay and flow cytometry were performed to detect cell proliferation and apoptosis of chondrocytes . The methylation level of the miR‑574‑5p promoter was measured via methylation specific PCR. The degree of chondrocyte apoptosis and the expression levels of YAF2 and Bcl‑2 were decreased in the mice with OA, and were increased in the OA + CTS mice, while the expression levels of miR‑574‑5p and Bax showed opposite changes. Furthermore, the degree of chondrocyte apoptosis and the expression levels of the aforementioned key factors in chondrocytes were consistent with those observed . The methylation degree of the miR‑574‑5p promoter was increased by the addition of CTS, and was reduced after the addition of a methylation inhibitor, 5‑aza‑CdR, indicating that CTS could regulate the methylation of miR‑574‑5p promoter. The present study suggested that CTS could downregulate the expression of miR‑574‑5p by regulating its methylation, and thus, could improve YAF2 expression and affect chondrocyte apoptosis.
软骨细胞凋亡是骨关节炎 (OA) 发展和进展的一个重要因素。隐丹参酮 (CTS) 可抑制软骨细胞凋亡,但具体机制尚不清楚。本研究旨在探讨 CTS 如何影响软骨细胞凋亡。采用逆转录-定量 PCR 和 Western blot 验证微小 RNA (miR)-574-5p、YY1 相关因子 2 (YAF2)、Bcl-2 和 Bax 表达水平。H&E、番红 O 和 TUNEL 染色检测关节炎软骨细胞凋亡。通过细胞计数试剂盒-8 检测和流式细胞术检测软骨细胞增殖和凋亡。采用甲基化特异性 PCR 检测 miR-574-5p 启动子的甲基化水平。OA 小鼠软骨细胞中 YAF2 和 Bcl-2 的表达水平降低,OA+CTS 小鼠中表达水平升高,miR-574-5p 和 Bax 的表达水平则相反。此外,软骨细胞中软骨细胞凋亡程度和上述关键因子的表达水平与观察结果一致。加入 CTS 后 miR-574-5p 启动子的甲基化程度增加,加入甲基化抑制剂 5-aza-CdR 后降低,表明 CTS 可调节 miR-574-5p 启动子的甲基化。本研究表明,CTS 可通过调节 miR-574-5p 的甲基化下调其表达,从而提高 YAF2 的表达,影响软骨细胞凋亡。