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目的观察黄芪甲苷对Wnt/β-catenin信号通路活化的人滑膜细胞和人软骨细胞共培养体系相关因子的调控作用,初步阐释黄芪甲苷治疗骨关节炎的分子作用机制。方法通过β-catenin慢病毒载体转染正常人滑膜细胞,激活其Wnt/β-catenin信号通路。Wnt/β-catenin信号通路活化的人滑膜细胞与正常人软骨细胞置于Transwell小室中共培养。黄芪甲苷混悬液干预共培养体系,采用ELISA方法检测黄芪甲苷对滑膜细胞、软骨细胞培养上清液基质金属蛋白酶-7(MMP-7)、Ⅱ型胶原羧基端肽(CTX-Ⅱ)、软骨寡聚基质蛋白(COMP)表达的影响。结果黄芪甲苷干预后,ELISA法检测发现滑膜与软骨细胞上清液中MMP-7、CTX-Ⅱ、COMP表达与正常对照相比明显升高(P<0.01)。结论黄芪甲苷可以抑制滑膜细胞Wnt/β-catenin信号通路,下调MMP-7、CTX-Ⅱ、COMP的表达。黄芪甲苷可能通过抑制骨关节炎滑膜Wnt/β-catenin信号通路,改善滑膜炎症、调控滑膜-软骨共同体系微环境、达到抑制软骨降解,促进软骨细胞功能恢复而治疗骨关节炎。
Objective To observe the regulatory effect of astragaloside IV on the related factors of co-culture system of human synovial cells and human chondrocytes activated by Wnt / β-catenin signaling pathway and to elucidate the molecular mechanism of astragaloside treatment of osteoarthritis. Methods The normal human synovial cells were transfected with β-catenin lentiviral vector to activate the Wnt / β-catenin signaling pathway. Human synoviocytes activated by Wnt / β-catenin signaling pathway were co-cultured with normal human chondrocytes in Transwell chambers. Astragaloside suspension was used to interfere with co-culture system. ELISA was used to detect the expression of matrix metalloproteinase-7 (MMP-7) and type Ⅱ collagen carboxyterminal peptide (CTX-Ⅱ) in synovial cells and chondrocytes ), Cartilage oligomeric matrix protein (COMP) expression. Results After astragaloside intervention, the expressions of MMP-7, CTX-Ⅱ and COMP in synovium and chondrocyte supernatant were significantly higher than those in normal control (P <0.01). Conclusion Astragaloside IV can inhibit Wnt / β-catenin signaling pathway and down-regulate the expression of MMP-7, CTX-Ⅱ and COMP in synoviocytes. Astragaloside IV may treat osteoarthritis by inhibiting the synovial Wnt / β-catenin signaling pathway of osteoarthritis, improving synovial inflammation, regulating the microenvironment of the synovial-cartilage common system, inhibiting the cartilage degradation and promoting the functional recovery of chondrocytes.