AMD3100体外阻断基质细胞衍生因子1/趋化因子受体4信号通路对人关节软骨细胞分泌基质金属蛋白酶3、9、13水平的影响

目的探讨趋化因子受体4(chemokine receptor 4,CXCR4)特异性拮抗剂AMD3100体外阻断基质细胞衍生因子1(stromal cell derived factor 1,SDF-1)/CXCR4信号通路对人关节软骨细胞分泌基质金属蛋白酶(matrixmetalloproteinase,MMP)3、9、13水平的影响,明确AMD3100的作用机制。方法取12例骨关节炎(osteoarthritis,OA)患者144块软骨组织(OA软骨组)和12例创伤性截肢患者144块正常软骨组织(正常软骨组)(Mankin评分均为0或1),根据添加培养液不同,每组再分为A、B、C 3个亚组。A亚组含1 000 nmol/L AMD3100及100 ng/mL SDF-1的DMEM液,B亚组含1 000 nmol/L MAB310及100 ng/mL SDF-1的DMEM液,C亚组含100 ng/mL SDF-1的DMEM液。体外培养2、4 d后,ELISA法测定培养液内MMP-3、9、13含量,RT-PCR检测软骨组织中MMP-3、9、13 mRNA表达。结果 ELISA法及RT-PCR检测示:同组相同时间点A亚组MMP-3、9、13含量及mRNA表达均显著低于B、C亚组(P<0.05)。同一时间点相同亚组OA软骨组MMP-3、9、13含量及mRNA表达均显著高于正常软骨组(P<0.05)。结论 SDF-1通过SDF-1/CXCR4信号通路可诱导人关节软骨中MMP-3、9、13的表达和释放;AMD3100可阻断SDF-1/CXCR4信号通路,使软骨细胞MMP-3、9、13 mRNA表达水平及分泌量降低,但AMD3100不能使退变的OA软骨分泌MMP-3、9、13恢复至正常软骨水平。

Influence on matrix metalloproteinases 3, 9, and 13 levels after blocking stromal cell derived factor 1/chemokine receptor 4 signaling pathway with AMD3100

Objective To investigate the in uence on matrix metalloproteinases(MMP) 3,9,and 13 levels of human articular cartilage cells after blocking stromal cell derived factor 1(SDF-1)/ chemokine receptor 4(CXCR4) signaling pathway with AMD3100 and to de ne the function mechanism of AMD3100.Methods A total of 144 cartilage blocks from 12 osteoarthritis(OA) patients undergoing total knee arthroplasty(OA cartilage group) and 144 normal cartilage blocks(Mankin score of 0 or 1) from 12 patients undergoing traumatic amputation(normal cartilage group).OA cartilage group was further divided into subgroups A1,B1,and C1,and normal cartilage group into subgroups A2,B2,and C2.The cartilage tissues were cultured in DMEM solution containing 100 ng/mL SDF-1 and 1 000 nmol/L AMD3100 in subgroup A,100 ng/mL SDF-1 and 1 000 nmol/L MAB310 in subgroup B,and 100 ng/mL SDF-1 in subgroup C,respectively.The levels of MMP-3,9,and 13 were measured by ELISA;the expressions of MMP-3,9,and 13mRNA were tested by RT-PCR.Results ELISA and RT-PCR results showed that the levels of MMP-3,9,and 13 and the expressions of MMP-3,9,and 13 mRNA were signi cantly lower in subgroup A than in subgroups B and C at the same time points(P < 0.05);the levels of MMP-3,9,and 13 and the expressions of MMP-3,9,and 13 mRNA were signi cantly higher in OA cartilage group than in normal cartilage group at the same time points(P < 0.05).Conclusion SDF-1 could induce overexpression and release of MMP-3,9,and 13 in the articular cartilage through the SDF-1/CXCR4 signaling pathway;AMD3100 could reduce the mRNA expressions and secretion of MMP-3,9,and 13 in OA cartilage by blocking the SDF-1/CXCR4 signaling pathway;but AMD3100 could not make the secretion of MMP-3,9,and 13 return to normal levels in OA cartilage.