miRNA-335-5p relieves chondrocyte inflammation by activating autophagy in osteoarthritis
miRNA-335-5p relieves chondrocyte inflammation by activating autophagy in osteoarthritis
复制标题
miRNA-335-5p 通过激活骨关节炎中的自噬减轻软骨细胞炎症
DOI：
10.1016/j.lfs.2019.03.071
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发表时间：
2019-06-01
期刊：
影响因子：
6.1
通讯作者：
Yao, Jun
中科院分区：
文献类型：
--
作者：
Zhong, Gang;Long, Huiping;Yao, Jun
目的：骨关节炎（Osteoarthritis，OA）是一种以关节软骨肥大和反应性增生为特征的老年慢性退行性关节疾病。自噬已被报道抑制炎症和减少OA中的软骨细胞凋亡。由于microRNA（miRNA）-335-5p与炎症和自噬有关，本研究旨在探讨其在OA发病机制中对自噬的调控作用。主要方法：采用实时定量PCR（qRT-PCR）检测正常和OA软骨细胞中miRNA-335- 5 p的表达。在用双链miRNA-335- 5 p模拟物/抑制剂转染人OA软骨细胞后，使用qRT-PCR、蛋白质印迹和免疫荧光来确定炎性介质IL-1 β、IL-6和TNF-α以及自噬标志物Beclin-1、自噬相关蛋白5（ATG 5）和ATG 7的表达水平。自噬抑制剂3-甲基腺嘌呤（3-MA）用于将miRNA-335- 5 p的抗炎作用与自噬联系起来。关键发现：OA软骨细胞中miRNA-335- 5 p的表达显著低于正常软骨细胞。用miRNA-335- 5 p模拟物转染人OA软骨细胞导致存活率显著增加，自噬相关因子显著增加，炎症介质减少。重要的是，用自噬抑制剂3-MA处理过表达miRNA-335 - 5 p的OA软骨细胞恢复了炎症介质的表达。因此，miRNA-335- 5 p在OA的临床诊断和治疗中具有潜在的应用前景。
Aims: Osteoarthritis (OA) is a chronic and degenerative joint disease prevalent in the elderly, which is characterized by hypertrophy and reactive hyperplasia of articular cartilage. Autophagy has been reported to inhibit inflammation and reduce chondrocyte apoptosis in OA. As the microRNA (miRNA)-335-5p has been linked to both inflammation and autophagy, this study aimed to investigate its potential role in regulating autophagy during the pathogenesis of OA.Main methods: Quantitative real-time PCR (qRT-PCR) was used to detect miRNA-335-5p expression in normal and OA human chondrocytes. Following transfection of human OA chondrocytes with double-stranded miRNA-335- 5p mimic/inhibitor, qRT-PCR, western blotting, and immunofluorescence were used to determine expression levels of the inflammatory mediators IL-1 beta, IL-6, and TNF-alpha, and the autophagic markers Beclin-1, autophagy-related protein 5 (ATG5), and ATG7. The autophagy inhibitor 3-methyladenine (3-MA) was used to link the anti-inflammatory effects of miRNA-335-5p to autophagy.Key findings: The expression of miRNA-335-5p was significantly lower in OA chondrocytes than in normal chondrocytes. Transfection of human OA chondrocytes with the miRNA-335-5p mimic led to a remarkable increase in viability, a significant increase in autophagy-related factors, and a reduction in inflammatory mediators. Importantly, treatment of miRNA-335-5p-overexpressing OA chondrocytes with the autophagy inhibitor 3-MA restored the expression of inflammatory mediators.Significance: We conclude that miRNA-335-5p can significantly alleviate inflammation in human OA chondrocytes by activating autophagy. Therefore, miRNA-335-5p has potential for future use in the clinical diagnosis and treatment of OA.