TMF通过上调SOCS3表达抑制骨关节炎软骨细胞凋亡和胞外基质降解

TMF protects against chondrocyte apoptosis and extracellular matrix degradation by activating SOCS3 expression in osteoarthritis

摘要

目的 骨关节炎（Osteoarthritis， OA）是一种以慢性炎症和软骨破坏为特征的骨病。软骨细胞是软骨中唯一的一种细胞类型，维持细胞外基质（Extracellular matrix， ECM）的稳态。软骨细胞功能障碍可破坏ECM稳态，诱导软骨降解，促进OA发展。细胞因子信号传导抑制剂3（Suppressor of cytokine signaling 3， SOCS3）在炎症中起着关键作用。本研究旨在探讨5，7，3'，4'-四甲氧基黄酮（5，7，3'，4'-tetramethoxyflavone， TMF）是否通过激活SOCS3表达来抑制软骨细胞凋亡和ECM降解。 方法 白介素-1β（Interleukin-1β， IL-1β）用于刺激C28/I2细胞建立OA样软骨细胞模型。采用流式细胞术检测软骨细胞凋亡。采用慢病毒感染敲减SOCS3表达。使用蛋白质印迹和免疫荧光分析来检测蛋白质表达。 结果 IL-1β可诱导软骨细胞凋亡，上调基质金属蛋白酶-13（Matrix metalloproteinase-13， MMP-13）表达，下调Ⅱ型胶原蛋白表达。此外，IL-1β可下调软骨细胞中SOCS3的表达。TMF能有效逆转IL-1β对软骨细胞的作用。SOCS3敲除可以消除TMF对IL-1β处理的软骨细胞的影响。 结论 TMF可通过激活SOCS3表达来抑制软骨细胞凋亡和ECM降解。

Abstract

Objective : Osteoarthritis （OA） is characterized by chronic inflammation and cartilage destruction. Chondrocyte， the unique cell type in the cartilage， is responsible for the maintenance of extracellular matrix （ECM） homeostasis. Chondrocyte dysfunction may disrupt ECM homeostasis， induce cartilage destruction， and promote OA development. Suppressor of cytokine signaling 3 （SOCS3） negatively plays a critical role in inflammation. This study aims to investigate whether 5，7，3'，4'-tetramethoxyflavone（TMF） inhibited chondrocyte apoptosis and ECM degradation by activating SOCS3 expression. Methods C28/I2 cells were stimulated by interleukin-1β （IL-1β） to establish OA-like chondrocyte models. Flow cytometry was used to detect chondrocyte apoptosis. Lentivirus infection was employed to knockdown SOCS3 expression. Western blotting and immunofluorescent assays were used to determine the protein expression. Results IL-1β treatment induced chondrocyte apoptosis， up-regulated matrix metalloproteinase-13 （MMP-13） expression， and down-regulated Collagen Ⅱ expression. In addition， the expression of SOCS3 was attenuated in IL-1β-treated chondrocytes. TMF could effectively reverse the effects of IL-1β on chondrocytes. SOCS3 knockdown could abolish the effects of TMF on IL-1β-treated chondrocytes. Conclusion TMF could inhibit OA chondrocyte apoptosis and suppress ECM degradation by activating SOCS3 expression.

关键词

骨关节炎 / 5，7，3'，4'-四甲氧基黄酮 / 软骨细胞凋亡 / 细胞外基质降解 / 细胞因子信号传导抑制剂3

Key words

Osteoarthritis / 5，7，3'，4'-tetramethoxyflavone / Chondrocyte apoptosis / Extracellular matrix degradation / Suppressor of cytokine signaling 3

基金资助

Funding： Project Supported by Jiangxi Provincial Natural Science Foundation基金项目(20212ACB206002)