[1]王春莉,梅虎,谢静,等.共培养下后交叉韧带成纤维细胞中MMPs的基因表达[J].陆军军医大学学报(原第三军医大学学报),2012,34(16):1600-1603.
 Wang Chunli,Mei Hu,Xie Jing,et al.Matrix metalloproteinase expression in posterior cruciate ligament fibroblasts co-cultured with synovial cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(16):1600-1603.
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第16期
页码:
1600-1603
栏目:
论著
出版日期:
2012-08-30

文章信息/Info

Title:
Matrix metalloproteinase expression in posterior cruciate ligament fibroblasts co-cultured with synovial cells
作者:
王春莉梅虎谢静蒋稼欢尹琳陈诚符纯锋KL Paul Sung
重庆大学生物工程学院国家“111计划”基地;美国加州大学圣迭亚哥分校骨科与生物工程系;重庆医科大学附属第一医院骨科
Author(s):
Wang Chunli Mei Hu Xie Jing Jiang Jiahuan Yin Lin Chen Cheng Fu Chunfeng KL Paul Sung
“111” Project Laboratory of Biomechanics and Tissue Repair, College of Bioengineering, Chongqing University, Chongqing, 400044, China; Department of Orthopedics and Bioengineering, University of California, San Diego, CA 9209320412, USA; Department of Orthopedics, First Affiliated Hospital, Chongqing Medical University, Chongqing, 400016, China
关键词:
后交叉韧带滑膜共培养基质金属蛋白酶
Keywords:
posterior cruciate ligament synovial co-culture matrix metalloproteinase
分类号:
R322.73;R329-33;R394.2
文献标志码:
A
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。
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.

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更新日期/Last Update: 2012-07-29