Soo C, Shaw W W, Zhang X, Longaker M T, Howard E W, Ting K
Department of Surgery, University of California at Los Angeles, 90095, USA.
Plast Reconstr Surg. 2000 Feb;105(2):638-47. doi: 10.1097/00006534-200002000-00024.
Wound extracellular matrix is a key regulator of cell adhesion, migration, proliferation, and differentiation during cutaneous repair. The amount and organization of normal wound extracellular matrix are determined by a dynamic balance among overall matrix synthesis, deposition, and degradation. Matrix metalloproteinases (MMPs) are one family of structurally related enzymes that have the collective ability to degrade nearly all extracellular matrix components. The MMPs are broadly categorized into collagenases, gelatinases, stromelysins, and membrane-type MMPs by their substrate specificity. The aim of this study was to characterize the temporal changes in mRNA profiles for rat collagenase [matrix metalloproteinase-1 (MMP-1)], gelatinase A (MMP-2), matrilysin (MMP-7), gelatinase B (MMP-9), and membrane type 1-MMP (MT1-MMP), as well as tissue inhibitor of metalloproteinases-1 (TIMP-1), TIMP-2, and TIMP-3 during the inflammatory, granulation, and early remodeling phases of excisional skin repair. Eight full-thickness skin wounds were made on the backs of each rat (7-mm2 wounds; 16 rats; n = 128 wounds). Two animals at a time were reanesthetized, and all eight wounds on each animal were excised at 12 and 24 hours and at 2, 3, 5, 7, 10, and 14 days after injury. Six wounds from each animal were excised for RNA isolation, whereas two wounds were excised for histology. Controls consisted of nonwounded skin from identical locations in four animals. Total RNA from each time point was isolated and relative mRNA quantitation performed by using reduced-cycle reverse transcription-polymerase chain reaction. Correct polymerase chain reaction product amplification was confirmed by probing the blotted polymerase chain reaction product with a 32P-labeled oligonucleotide specific for a given MMP or TIMP. We demonstrated that the majority of MMP and TIMP mRNA induction and peak expression coincided temporally with the well-characterized inflammatory and granulation stages of repair. In conclusion, there is a distinct pattern of MMP and TIMP expression during normal excisional wound repair.
伤口细胞外基质是皮肤修复过程中细胞黏附、迁移、增殖和分化的关键调节因子。正常伤口细胞外基质的数量和组织由整体基质合成、沉积和降解之间的动态平衡决定。基质金属蛋白酶（MMPs）是一类结构相关的酶，它们共同具有降解几乎所有细胞外基质成分的能力。根据底物特异性，MMPs大致分为胶原酶、明胶酶、基质溶解素和膜型MMPs。本研究的目的是表征大鼠胶原酶[基质金属蛋白酶-1（MMP-1）]、明胶酶A（MMP-2）、基质溶素（MMP-7）、明胶酶B（MMP-9）和膜型1-MMP（MT1-MMP）以及金属蛋白酶组织抑制剂-1（TIMP-1）、TIMP-2和TIMP-3在切除性皮肤修复的炎症、肉芽组织形成和早期重塑阶段的mRNA谱的时间变化。在每只大鼠的背部制作8个全层皮肤伤口（7平方毫米伤口；16只大鼠；n = 128个伤口）。每次对两只动物重新麻醉，并在受伤后12小时和24小时以及2、3、5、7、10和14天切除每只动物身上的所有8个伤口。从每只动物身上切除6个伤口用于RNA分离，而切除2个伤口用于组织学检查。对照组由来自4只动物相同位置的未受伤皮肤组成。从每个时间点分离总RNA，并使用减少循环逆转录-聚合酶链反应进行相对mRNA定量。通过用针对给定MMP或TIMP的32P标记寡核苷酸探测印迹的聚合酶链反应产物来确认正确的聚合酶链反应产物扩增。我们证明，大多数MMP和TIMP mRNA的诱导和峰值表达在时间上与特征明确的修复炎症和肉芽组织形成阶段一致。总之，在正常切除性伤口修复过程中，MMP和TIMP表达存在明显模式。
