通络生骨胶囊对软骨氧化损伤的保护作用及其机制探讨

Antioxidation and Mechanism of Tongluo Shenggu Capsule on Oxidative Damage of Cartilage

目的:通过H2O2诱导(SW1353)软骨细胞损伤Model,研究通络生骨胶囊对软骨细胞的保护作用,并探索其可能的作用机制。方法:不同浓度的通络生骨胶囊(1、5、10、50和100μg/mL)预处理SW1353细胞72 h,再经H 2 O 2(300μmol/L)诱导损伤1 h,检测细胞活率、Nrf2、HO-1、Wnt4α和β-catenin的mRNA表达,并检测β-catenin、Nrf2、Keap-1和HO-1的蛋白表达水平。结果:通络生骨胶囊可有效促进软骨细胞增殖;在H 2 O 2损伤Model中,通络生骨胶囊对软骨细胞具有明显的抗氧化保护作用,同时增强Nrf2、HO-1、Wnt4α和β-catenin的mRNA表达水平。通络生骨胶囊还可有效抑制Keap-1蛋白表达,上调Nrf2和HO-1的蛋白表达。结论:通络生骨胶囊可能通过抑制Wnt/β-catenin信号通路从而激活Nrf2/HO-1抗氧化途径,降低氧自由基对软骨细胞损伤作用,从而达到治疗骨性关节炎的目的。

Objective:To evaluate the antioxidative actions of Tongluo Shenggu Capsule on chondrocytes(SW1353)and its feasible mechanisms.Methods:The chondrocytes were injured by H 2O 2(300μmol/L)for 1 h after being pre-treated by various concentrations of Tongluo Shenggu Capsule(1μg/mL,5μg/mL,10μg/mL,50μg/mL and 100μg/mL)for 72 h,then cell viability,the mRNA expressions of Nrf2,HO-1,Wnt4αandβ-catenin,as well as the protein expressions ofβ-catenin,Nrf2,Keap-1 and HO-1 were detected.Results:Tongluo Shenggu Capsule could effectively promote the proliferation of chondrocytes.In the H 2O 2-damage model,Tongluo Shenggu Capsule had obviously antioxidant action,and enhanced the mRNA expressions of Nrf2,HO-1,Wnt4αandβ-catenin.It could also effectively inhibit Keap-1 protein expression and up-regulate protein expressions of Nrf2 and HO-1.Conclusion:Tongluo Shenggu Capsule could reduce the damage of oxygen free radicals to chondrocytes,and activate Nrf2/HO-1 pathway by inhibiting Wnt/β-catenin signaling pathway,so as to achieve the purpose of treating osteoarthritis.