【摘要】 目的研究后交叉韧带(posterior cruciate ligament,PCL)成纤维细胞与滑膜细胞共培养下,2种细胞之间的交流对PCL成纤维细胞中基质金属蛋白酶MMP-1、2、3表达及活性的影响。方法分为共培养组和单培养组:①用显微镜观察共培养24 h后细胞的活性和迁移率。②分别培养6 h后,提取总RNA,逆转录PCR;实时荧光定量PCR对人后交叉韧带成纤维细胞MMP-1、2、3基因的表达进行半定量和定量分析。③共培养处理1、2、3 d后收集培养上清液,用明胶酶谱法检测MMP-2的活性。结果共培养使PCL细胞和滑膜细胞较单层细胞培养迁移率分别增高了(43.1±0.1)%和(76.2±0.2)%(P<0.01)。与单培养相比,共培养组明显增加了MMP-2的基因表达,但降低了MMP-1、3的基因表达。酶谱结果表明共培养增加了MMP-2活性(P<0.05)。结论共培养能够促进细胞的迁移以及调节细胞中MMP-1、2、3的表达情况。更多还原

【Abstract】 Objective To investigate the expression of matrix metalloproteinase(MMP)-1,MMP-2 and MMP-3 in posterior cruciate ligament(PCL) fibroblasts co-cultured with synovial cells.Methods Human PCL fibroblasts were co-cultured with synovial cells and the monocultures of human PCL fibroblasts and synovial cells were assigned as control.Cell viability and migration were observed by microscopy after 24 h incubation.Total RNA was isolated after 6 h incubation and the expression of MMPs was analyzed by semi-quantitative PCR and quantitative real-time PCR.Gelatin zymography was applied to detect the MMP-2 activity in culture supernatants collected at 1,2 and 3 d after incubation.Results As compared with the monocultures,the migration rates of the human PCL fibroblasts and synovial cells in the co-culture increased by(43.1±0.1)% and(76.2±0.2)%,respectively(P<0.01).The mRNA expression levels of MMP-1 and MMP-3 in the PCL fibroblasts co-cultured with synovial cells were lower than those in the PCL fibroblasts monoculture.However,PCL fibroblasts-synovial cells interaction significantly increased the expression and activity of MMP-2 in the PCL fibroblasts(P<0.05).Conclusion The differential expression of MMP-1,MMP-2 and MMP-3 in human PCL fibroblasts co-cultured with synovial cells indicates that cell-cell interaction and cross-talking are associated with PCL fibroblasts wound healing and have potential clinical value for curing injured PCL.更多还原