GSK-3β/Wnt双信号调控对炎症环境关节软骨损伤修复的作用

Athletic Injuries or chronic inflammatory diseases can induce injury of articular cartilage, which has poor capacity for self-repair and regeneration in inflammatory environment. The balance between chondrogenic differentiations promoted by cytokine and depressed by inflammation should be focused. GSK-3βcould regulates cell proliferation, differentiation and apoptosis virtually. It was found that GSK-3β could regulate NF-κB pathway in tumor cells. We also verified cytokines of NF-κB pathway could recruit BMSCs migration from bone marrow to one body site where they became maturation. Wnt signaling plays a crucial role in chondrogenesis through Wnt/β-catenin pathway. Hence, We have the hypothesis that there is interaction with GSK-3β and Wnt signaling. It can not only control activity of inflammation and regulate its inflammatory effect to chondrogenesis process, but also can co-regulate BMSCs to migrate to cartilage injury site and induce cartilage restoration. This study would focus on investigating the role of GSK-3β and Wnt double signal regulation to chondrocytes and BMSCs proliferation, differentiation and apoptosis, verifying the inereaction and figure out the mechanism in cartilage restoration of this kind feedback effects by animal experiments. It may provide an alternative theoretical support to study the process of cartilage damage and restoration and its clinical treatment.

运动损伤或慢性炎症疾病常导致关节软骨损伤，但炎症环境下关节软骨损伤自我修复能力低下，如何调节细胞因子促软骨再生和炎症作用的平衡是研究的重点。GSK-3β广泛调节细胞的增殖、分化和凋亡，最新研究发现肿瘤细胞中GSK-3β可以调控NF-κB炎症通路活性，申请人参与的研究证实炎症信号可以诱导BMSCs从骨髓定向迁移至成熟分化的部位。Wnt可通过Wnt/β-catenin信号参与软骨增殖分化的调控。故提出猜想，GSK-3β和Wnt信号是否能协同作用既降低微环境中NF-κB信号对成软骨分化不利影响，又能发挥其促BMSCs募集定向迁移至软骨损伤部位进行软骨修复的作用？本课题研究GSK-3β和Wnt双信号对软骨细胞和BMSCs增殖、凋亡和成软骨分化的影响，探索二者之间是否存在交互反馈作用以及这种作用对炎症环境软骨损伤修复的影响机制，通过动物实验证实，为研究软骨损伤修复过程和治疗方案提供理论支持。

运动损伤或慢性炎症疾病常导致人体负重关节的关节面软骨损伤，但体内软骨损伤自我修复能力低下。研究炎症环境中一些信号因子对软骨再生和炎症的作用成为热点。GSK-3β广泛调节细胞的增殖、分化和凋亡，研究发现肿瘤细胞中GSK-3β可以调控NF-κB炎症通路活性，而Wnt可通过Wnt/β-catenin信号参与软骨增殖分化的调控。本研究认为GSK-3β和Wnt信号可能有协同作用，通过对软骨细胞和BMSCs分别调控上述信号，期待找到理想的条件，既能降低炎症环境NF-κB信号对成软骨分化的抑制，又能提高软骨细胞或BMSCs促进软骨损伤修复过程。.本项目是以软骨细胞和BMSCs为研究对象，分别上调和下调GSK-3β信号、上调和下调Wnt信号，研究软骨细胞相关细胞表型蛋白GAG/Aggrecan/Collagen II/Sox9的表达情况，检测内源性GSK-3β，磷酸化p-GSK-3β水平。检测NF-κB/p65因子在细胞内的总体水平、磷酸化入核水平。检测胞浆β-catenin 和磷酸化水平。检测凋亡相关基因如Caspase9、Caspase3、Bax、Bcl-2、Surivin 的表达情况。.本研究实验结果符合课题假设。总体趋势趋势是，在炎症环境下软骨细胞的表型受到抑制。在软骨细胞和BMSCs中，抑制GSK-3β信号能促进软骨细胞表型的表达，上调GSK-3β信号，则观察到相反的结果；在抑制Wnt信号的情况下，BMSCs成软骨分化过程受到抑制。这与同期其他研究结果的趋势基本一致。.本课题通过探讨GSK-3β和Wnt信号对软骨细胞表型相关蛋白的影响和对BMSCs成软骨分化过程的作用，旨在找出有利于软骨损伤修复和软骨细胞再生的有利条件，为下一步开展组织工程技术进行体内软骨损伤修复试验奠定理论依据。

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