T140体外阻断SDF-1/CXCR4信号通路阻止人关节软骨Ⅱ型胶原蛋白降解的研究

In vitro blockade of SDF-1/CXCR4 signaling pathway by T140 to prevent degradation of type Ⅱ collagen in human articular cartilage

目的:探讨T140体外阻断基质细胞衍生因子1(stromal cell derived factor 1,SDF-1)/趋化因子受体4(chemokine receptor 4,CXCR4)信号通路对人关节软骨细胞降解Ⅱ型胶原蛋白的影响,明确T140的作用机制。方法:取144块膝关节置换OA患者软骨(OA软骨组)和144块创伤性截肢患者正常软骨(正常软骨组)组织,Mankin评分均为0或1,加入SDF-1(100 ng/mL)。每组再分为A、B、C三个亚组,分别加入浓度为1 000 nmol/L的T140、MAB310和SDF-1,分别于体外培养2、4 d后,采用RT-PCR检测软骨组织Ⅱ型胶原蛋白mRNA表达。结果:相同软骨组在相同时间点,A组Ⅱ型胶原蛋白mRNA表达均显著高于B组和C组(P<0.05)。相同时间点,OA软骨组同一亚组Ⅱ型胶原蛋白mRNA表达均显著低于正常软骨组(P<0.05)。结论:SDF-1通过SDF-1/CXCR4信号通路诱导人关节软骨降解Ⅱ型胶原蛋白;T140阻断SDF-1/CXCR4信号通路,缓解软骨细胞降解Ⅱ型胶原蛋白。

Objective To explore the impact of blockade of SDF-1/CXCR4 signaling pathway by T140 on degradation of type Ⅱ collagen in human articular cartilage and to define the mechanism of action of T140. Methods 144 pieces of cartilage （Mankin score of 0 or 1 ） were obtained from osteoarthritis patients undergoing total knee replacement （ OA cartilage group） and 144 pieces of cartilage （Mankin score of 0 or 1） were obtained from patients receiving traumatic amputation （normal cartilage group）. Each group was treated with SDF-1 of 100 ng/mL, then divided into three subgroups A, B, and C. The cartilage tissue in each group was cultured in the nutrient solution containing of T140, MAB310, or SDF-1 （ 1 000 nmol/L） for 48 or 96 hours. RT-PCR was used to detect expression of type Ⅱ collagen mRNA in the cartilage tissue. Results Level of type Ⅱ collagen mRNA was markedly higher in subgroup A than in subgroup B and subgroup C at the same group and the same time （P 〈 0.05）. The expression level of type Ⅱ collagen mRNA at the same time and in the same subgroup of OA cartilage group were lower than that in normal cartilage group （P 〈 0.05）. Conclusions SDF-1 induces degradation of type Ⅱ collagen in human articular cartilage thruogh the SDF-1/CXCR4 signaling pathway. T140 can block the SDF-1/ CXCR4 signaling pathway and reduce the degradation of type Ⅱ collagen.