TGF-beta1 induces the different expressions of lysyl oxidases and matrix metalloproteinases in anterior cruciate ligament and medial collateral ligament fibroblasts after mechanical injury.
TGF-beta1 induces the different expressions of lysyl oxidases and matrix metalloproteinases in anterior cruciate ligament and medial collateral ligament fibroblasts after mechanical injury.
复制标题
DOI：
10.1016/j.jbiomech.2012.12.019
复制
发表时间：
2013-03
影响因子：
2.4
通讯作者：
Jing Xie;Chunli Wang;Dongyue Huang;Yanyan Zhang;Jianwen Xu;S. Kolesnikov;K. Sung;Hu-cheng Zhao
中科院分区：
文献类型：
--
作者：
Jing Xie;Chunli Wang;Dongyue Huang;Yanyan Zhang;Jianwen Xu;S. Kolesnikov;K. Sung;Hu-cheng Zhao
众所周知，前交叉韧带（ACL）具有较差的自愈能力。相比之下，内侧副韧带（MCL）可以较好地愈合并恢复关节功能。转化生长因子-β1 （TGF-β1）被认为是韧带创面愈合的重要化学介质。而TGF-β1诱导的赖氨酸氧化酶（LOXs）和基质金属蛋白酶（MMPs）的表达，分别促进细胞外基质（ECM）的修复和降解，其作用尚不清楚。本研究采用等双轴拉伸室模拟ACL和MCL成纤维细胞的机械损伤，旨在通过表征LOXs和MMPs在TGF-β1作用下的机械损伤后表达差异来确定ACL和MCL的内在差异。通过半定量PCR、实时定量PCR、western blot和酶谱分析，我们发现TGF-β1诱导损伤的MCL比损伤的ACL表达更多的LOXs （LOX为1.85倍，LOXL-1为2.21倍，LOXL-2为1.71倍，LOXL-3为2.52倍，LOXL-4为3.32倍）。同时，TGF-β1诱导损伤的前交叉韧带比损伤的MCL成纤维细胞表达更多的MMPs （MMP-1表达2.33倍，MMP-2表达2.45倍，MMP-3表达1.89倍，MMP-12表达1.50倍）。进一步的蛋白表达结果与上述基因表达一致。LOXs和MMPs的不同表达推断了ACL和MCL之间的内在差异，而内在差异可以帮助解释它们不同的愈合能力。
The anterior cruciate ligament (ACL) is known to have a poor self-healing ability. In contrast, the medial collateral ligament (MCL) can heal relatively well and restore the joint function. Transforming growth factor-beta1 (TGF-β1) is considered to be an important chemical mediator in the wound healing of the ligaments. While the role of TGF-β1-induced expressions of the lysyl oxidases (LOXs) and matrix metalloproteinases (MMPs), which respectively facilitate the extracellular matrix (ECM) repair and degradation, is poorly understood. In this study, we used equibiaxial stretch chamber to mimic mechanical injury of ACL and MCL fibroblasts, and aimed to determine the intrinsic differences between ACL and MCL by characterizing the differential expressions of LOXs and MMPs in response to TGF-β1after mechanical injury. By using semi-quantitative PCR, quantitative real-time PCR, western blot and zymography, we found TGF-β1induced injured MCL to express more LOXs than injured ACL (up to 1.85-fold in LOX, 2.21-fold in LOXL-1, 1.71-fold in LOXL-2, 2.52-fold in LOXL-3 and 3.32-fold in LOXL-4). Meanwhile, TGF-β1induced injured ACL to express more MMPs than injured MCL fibroblasts (up to 2.33-fold in MMP-1, 2.45-fold in MMP-2, 1.89-fold in MMP-3 and 1.50-fold in MMP-12). The further protein results were coincident with the gene expressions above. The different expressions of LOXs and MMPs inferred the intrinsic differences between ACL and MCL, and the intrinsic differences could help to explain their differential healing abilities.