MMP-2 mediates local degradation and remodeling of collagen by annulus fibrosus cells of the intervertebral disc.
MMP-2 mediates local degradation and remodeling of collagen by annulus fibrosus cells of the intervertebral disc.
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DOI：
10.1186/ar4224
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发表时间：
2013-04-27
影响因子：
4.9
通讯作者：
Hsieh AH
中科院分区：
文献类型：
--
作者：
Rastogi A;Kim H;Twomey JD;Hsieh AH
椎间盘退行性变(IVD)的特征是纤维环(AF)的显著降解和重建。尽管已发现几种基质金属蛋白酶(MMPs)在退变的椎间盘中更为普遍，但它们在疾病过程中的协调和功能仍不清楚。在这项研究中，我们试图确定基质金属蛋白酶-2是否与房颤的退行性改变有关，并确定房颤细胞使用基质金属蛋白酶-2的方式。对两种已建立的椎间盘退变动物模型--静态压迫和跨环针穿刺鼠尾盘模型进行了检测，以检测基质金属蛋白酶-2免疫阳性反应。通过慢病毒转导shRNA表达盒，我们筛选和鉴定了一个有效的shRNA序列，以产生稳定的RNA干扰来沉默原代大鼠房颤细胞中基质金属蛋白酶-2的表达。明胶薄膜被用来比较转导细胞和未感染和无意义shRNA对照细胞之间的明胶酶活性和降解的空间模式。通过评估细胞重塑胶原胶的能力来确定基质金属蛋白酶-2的功能意义。静态加压和18g环状穿刺法均可刺激房颤中基质金属蛋白酶-2活性增加，同时伴有板层破坏，而22g和26g针刺伤则不能。为了研究基质金属蛋白酶-2的功能作用，我们建立了慢病毒介导的RNAi，在10天内诱导转录水平稳定下调88%，蛋白质水平稳定下调95%。在明胶膜上培养转导细胞证实了基质金属蛋白酶-2是房颤细胞的主要功能明胶酶，并且在房颤细胞周围的区域局部使用了基质金属蛋白酶-2。在胶原凝胶中，转导细胞表现出不能重塑胶原基质。我们的研究表明，在人类退变的椎间盘中观察到的基质金属蛋白酶-2的增加反映在实验诱导的啮齿动物模型的退行性变化中。房颤细胞似乎以一种非常直接的方式使用基质金属蛋白酶-2来进行局部基质降解和胶原重塑。这表明，基质金属蛋白酶-2在退行性腰椎间盘疾病的发病机制中可能起着重要的作用，并可能成为潜在的治疗靶点。
Degeneration of the intervertebral disc (IVD) is characterized by marked degradation and restructuring of the annulus fibrosus (AF). Although several matrix metalloproteinases (MMPs) have been found to be more prevalent in degenerate discs, their coordination and function within the context of the disease process are still not well understood. In this study, we sought to determine whether MMP-2 is associated with degenerative changes in the AF and to identify the manner by which AF cells use MMP-2. Two established animal models of disc degeneration, static compression and transannular needle puncture of rodent caudal discs, were examined for MMP-2 immunopositivity. With lentiviral transduction of an shRNA expression cassette, we screened and identified an effective shRNA sequence for generating stable RNA interference to silence MMP-2 expression in primary rat AF cells. Gelatin films were used to compare gelatinase activity and spatial patterns of degradation between transduced cells, and both noninfected and nonsense shRNA controls. The functional significance of MMP-2 was determined by assessing the ability for cells to remodel collagen gels. Both static compression and 18-g annular puncture of rodent caudal discs stimulated an increase in MMP-2 activity with concurrent lamellar disorganization in the AF, whereas 22-g and 26-g needle injuries did not. To investigate the functional role of MMP-2, we established lentivirus-mediated RNAi to induce stable knockdown of transcript levels by as much as 88%, and protein levels by as much as 95% over a 10-day period. Culturing transduced cells on gelatin films confirmed that MMP-2 is the primary functional gelatinase in AF cells, and that MMP-2 is used locally in regions immediately around AF cells. In collagen gels, transduced cells demonstrated an inability to remodel collagen matrices. Our study indicates that increases in MMP-2 observed in human degenerate discs are mirrored in experimentally induced degenerative changes in rodent animal models. AF cells appear to use MMP-2 in a very directed fashion for local matrix degradation and collagen remodeling. This suggests that MMP-2 may have a functionally significant role in the etiology of degenerative disc disease and could be a potential therapeutic target.
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影响因子：
4.3
作者：
Lieu S;Hansen E;Dedini R;Behonick D;Werb Z;Miclau T;Marcucio R;Colnot C
通讯作者：
Colnot C
影响因子：
--
作者：
Aigner, Achim
通讯作者：
Aigner, Achim
影响因子：
4.8
作者：
AIMES, RT;QUIGLEY, JP
通讯作者：
QUIGLEY, JP
DOI：
10.1083/jcb.136.6.1363
发表时间：
1997-03-24
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lelongt B;Trugnan G;Murphy G;Ronco PM
通讯作者：
Ronco PM
影响因子：
3.7
作者：
Deryugina, EI;Ratnikov, B;Strongin, AY
通讯作者：
Strongin, AY