BACKGROUND: Osteoarthritis is a common cause of pain and disability in the elderly. The underlying cause is the combination of inappropriate mechanical stress, inflammatory mediators and biochemical factors. Curcumin has been shown to have anti-oxidant, anti-inflammatory, anti-bacterial, anti-tumor, neuroprotective, and cardioprotective effects, radiation protection and therapeutic effects on osteoarthritis. Similarly, magnesium sulfate can relieve joint pain and inhibit joint destruction. OBJECTIVE: To investigate the mechanism by which the combined use of curcumin and magnesium sulfate exerts synergistic effect to promote inflammatory chondrocyte reverse differentiation.METHODS: Primary cultured inflammatory chondrocytes were subjected to monolayer culture in vivo. Cell proliferation assay (MTS) was used to detect the proliferation of inflammatory chondrocytes under in vitro monolayer culture conditions. The experimental cells were divided into four groups and underwent 3D induced reverse differentiation culture for 18 days: single culture of chondrocytes (chondrocyte group) , inflammatory chondrocyte cultured with curcumin (curcumin group) , inflammatory chondrocytes cultured with magnesium sulfate (magnesium sulfate group) , and inflammatory chondrocytes cultured with curcumin and magnesium sulfate (combination group). RESULTS AND CONCLUSION: MTS proliferation experiments showed that inflammatory chondrocytes at passage 3 had a higher rate of early proliferation and a lower degree of differentiation. Quantitative PCR results showed that the mRNA levels of type II collagen, proteoglycan and SOX9 were significantly higher in the combination group than in the curcumin group or magnesium sulfate group (P < 0.01). The size of the gross specimens and the positive area of chondrocyte reverse differentiation for alcian blue staining and safranin O staining in the combination group were significantly larger than those in the other three groups (P < 0.01). TUNEL staining results indicated that the positive area of apoptosis-specific staining in the combination group and magnesium sulfate group was significantly smaller than that in the other two groups (P < 0.01). Therefore, the combined use of curcumin and magnesium sulfate has the synergistic effect to promote the reverse differentiation of inflammatory chondrocytes.%背景:骨性关节炎是老年人疼痛和致残的常见原因, 其根本原因在于不适当的机械应力、炎症递质和生化因子的共同作用.姜黄素已被证明具有抗氧化、抗炎、抗菌、抗肿瘤、神经保护、心脏保护、辐射防护以及治疗骨关节炎等作用, 硫酸镁 (MgSO4) 同样具有缓解关节疼痛, 促进关节破坏的作用.目的:探究姜黄素和MgSO4联合应用协同促进炎性软骨细胞逆分化的作用机制.方法:原代培养炎性软骨细胞, 在体外单层培养条件下, 细胞增殖实验检测炎性软骨细胞的增殖情况.实验细胞分为4组, 进行体外3D诱导逆分化培养, 炎性软骨细胞组为炎性软骨细胞单独培养, 姜黄素组采用姜黄素干预培养, MgSO4组为采用MgSO4干预培养, 联合治疗组为姜黄素+MgSO4联合干预培养, 共培养18 d.结果与结论: (1) 细胞增殖实验显示, 第3代炎性软骨细胞早期增殖速率较高, 分化程度较低; (2) 定量PCR检测显示, 联合治疗组Ⅱ型胶原、蛋白聚糖和SOX9 mRNA表达量均高于姜黄素组或MgSO4组 (P <0.01) ; (3) 体外3D诱导分化培养中联合治疗组的大体标本的尺寸和软骨逆分化的阿尔新蓝染色、番红O染色的阳性区域均明显高于其他3组 (P <0.01) ; (4) TUNEL染色显示, 联合治疗组及MgSO4组凋亡特异性染色的阳性区域均明显小于另外2组 (P <0.01) ; (5) 结果证实, 联合应用姜黄素和MgSO4具有协同促进炎性软骨细胞逆分化的作用.
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