共培养下后交叉韧带成纤维细胞中MMPs的基因表达  

Matrix metalloproteinase expression in posterior cruciate ligament fibroblasts co-cultured with synovial cells

作　　者：王春莉[1] 梅虎[1] 谢静[1] 蒋稼欢[1] 尹琳[1] 陈诚[3] 符纯锋[3] KL Paul Sung 

机构地区：[1]重庆大学生物工程学院国家"111计划"基地,400044 [2]美国加州大学圣迭亚哥分校骨科与生物工程系,美国圣迭亚哥9209320412 [3]重庆医科大学附属第一医院骨科,400016

出　　处：《第三军医大学学报》2012年第16期1600-1603,共4页Journal of Third Military Medical University

基　　金：高等学校学科创新引智计划(国家“111计划”)(B06023);重庆市科技攻关项目(CSTC2008AB5)~~

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。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 super- natants collected at 1,2 and 3 d after incubation. Results As compared with the monocuhures, the migration rates of the human PCL fibroblasts and synovial ceils 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 fi- broblasts co-cultured with synovial cells were lower than those in the PCL fibroblasts monocuhure. However, PCL fibroblasts-synovial cells interaction significantly increased the expression and activity of MMP-2 in the PCL fibroblasts （P 〈 0. 05 ）. Concausion 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.

关 键 词：后交叉韧带 滑膜 共培养 基质金属蛋白酶 

分 类 号：R322.73[医药卫生—人体解剖和组织胚胎学] R329.33[医药卫生—基础医学]