臭氧通过激活 PPARγ/mTOR 诱导 IL-1β 处理的大鼠软骨细胞自噬。
Ozone induces autophagy by activating PPARγ/mTOR in rat chondrocytes treated with IL-1β.
机构信息
Department of Pain Management, The Second Hospital of Shandong University, Jinan, 250033, People's Republic of China.
Department of Pain Management, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, 250021, People's Republic of China.
出版信息
J Orthop Surg Res. 2022 Jul 16;17(1):351. doi: 10.1186/s13018-022-03233-y.
BACKGROUND
Osteoarthritis (OA) is the main cause of older pain and disability. Intra-articular injections of ozone (O) commonly have been found to have antioxidative and anti-inflammatory effects to reduce pain and improve function in knee osteoarthritis. It has been reported that reduced autophagy in chondrocytes plays an important role in the development of OA. This study aimed to probe the role of O on the autophagy in chondrocytes treated with IL-1β.
METHODS
Primary chondrocytes were isolated from Wistar rats cartilage within 3 days. The OA chondrocytes model was induced via treatment with IL-1β for 24 h. Then the cells were treated with O and GW9662, the inhibitor of PPARγ. Cell viability was assessed by CCK-8. Further, the cells subjected to Western blot analysis, qRT-PCR and immunofluorescence assay. The numbers of autophagosomes were observed via transmission electron microscopy.
RESULTS
30 μg/ml O improved the viability of chondrocytes treated with IL-1β. The decreased level of autophagy proteins and the numbers of autophagosomes improved in IL-1β-treated chondrocytes with O via activating PPARγ/mTOR. In addition, the qRT-PCR results showed that O decreased the levels of IL-6, TNF-α and MMP-3, MMP-13 in chondrocytes treated with IL-1β.
CONCLUSIONS
30 μg/ml O improved autophagy via activating PPARγ/mTOR signaling and suppressing inflammation in chondrocytes treated with IL-1β.
背景
骨关节炎（OA）是老年人疼痛和残疾的主要原因。关节内注射臭氧（O）通常被发现具有抗氧化和抗炎作用，可减轻膝关节骨关节炎的疼痛并改善功能。据报道，软骨细胞中的自噬减少在 OA 的发展中起重要作用。本研究旨在探讨 O 对 IL-1β 处理的软骨细胞自噬的作用。
方法
从 Wistar 大鼠软骨中分离出原代软骨细胞，在 3 天内。通过用 IL-1β 处理 24 小时来诱导 OA 软骨细胞模型。然后用 O 和 GW9662（PPARγ 的抑制剂）处理细胞。通过 CCK-8 评估细胞活力。进一步进行 Western blot 分析、qRT-PCR 和免疫荧光测定。通过透射电子显微镜观察自噬体的数量。
结果
30μg/ml O 提高了 IL-1β 处理的软骨细胞的活力。通过激活 PPARγ/mTOR，O 改善了 IL-1β 处理的软骨细胞中自噬蛋白水平和自噬体数量的降低。此外，qRT-PCR 结果表明，O 降低了 IL-1β 处理的软骨细胞中 IL-6、TNF-α 和 MMP-3、MMP-13 的水平。
结论
30μg/ml O 通过激活 PPARγ/mTOR 信号通路改善自噬，并抑制 IL-1β 处理的软骨细胞中的炎症。
Zotero 插件