Wu Nanjun, Davidson Jeffrey M
Department of Pathology, Vanderbilt University School of Medicine, Medical Center, Nashville, TN 37232-2561, USA.
Wound Repair Regen. 2004 May-Jun;12(3):327-36. doi: 10.1111/j.1067-1927.2004.012313.x.
Incisions made in mouse skin by scalpel or the free-electron laser heal at different rates. To identify genes that are differentially expressed in free-electron laser or scalpel wounds, we isolated total RNA from free-electron laser- or scalpel-produced incisions and normal skin at day 7 postwounding. cDNA microarray analysis identified 89 of 15,000 genes in a mouse microarray as having significantly different expression levels. Migration inhibitory factor-related protein (MRP) 14 was almost 30 times more highly expressed in scalpel wounds than in free-electron laser wounds. This result was confirmed by Northern blot analysis, which also showed that scalpel wounds expressed higher levels of MRP8, a related S100 protein that can heterodimerize with MRP14, at days 2, 7, and 14 postwounding. Free-electron laser wounds also showed elevated expression of MRP8 and MRP14 relative to normal skin. In situ hybridization showed that the patterns of MRP14 and MRP8 expression in free-electron laser and scalpel wound tissues were similar. MRP14 and MRP8 were expressed in the dermal wound margin, while a very low level of MRP14 and MRP8 expression was seen in the migrating epidermis. Dual immunofluorescence staining for MRP14 or MRP8 and macrophage (F4/80) showed that most of the wound macrophages simultaneously expressed MRP14 and MRP8. Some expression was also found in neutrophils, while neither antigen accumulated to a significant degree in the epidermis. Relatively lower MRP8 and 14 expression in free-electron laser wounds was correlated with a higher level of matrix metalloproteinase-13 expression and a reduced rate of wound healing. While the regulation of MRP8 expression in mouse may be different from human skin, we suggest that elevated expression of MRP8 and MRP14 may have a relevant therapeutic effect against inflammation in wound healing.
用手术刀或自由电子激光在小鼠皮肤上造成的切口愈合速度不同。为了鉴定在自由电子激光或手术刀造成的伤口中差异表达的基因,我们在受伤后第7天从自由电子激光或手术刀造成的切口以及正常皮肤中分离了总RNA。cDNA微阵列分析确定,在小鼠微阵列的15000个基因中,有89个基因的表达水平存在显著差异。迁移抑制因子相关蛋白(MRP)14在手术刀造成的伤口中的表达量几乎是自由电子激光伤口中的30倍。Northern印迹分析证实了这一结果,该分析还表明,在受伤后第2、7和14天,手术刀造成的伤口中MRP8(一种可与MRP14形成异二聚体的相关S100蛋白)的表达水平更高。相对于正常皮肤,自由电子激光伤口中MRP8和MRP14的表达也有所升高。原位杂交显示,自由电子激光和手术刀伤口组织中MRP14和MRP8的表达模式相似。MRP14和MRP8在真皮伤口边缘表达,而在迁移的表皮中MRP14和MRP8的表达水平非常低。对MRP14或MRP8与巨噬细胞(F4/80)进行双重免疫荧光染色显示,大多数伤口巨噬细胞同时表达MRP14和MRP8。在中性粒细胞中也发现了一些表达,而两种抗原在表皮中均未大量积累。自由电子激光伤口中相对较低的MRP8和14表达与基质金属蛋白酶-13的较高表达水平以及伤口愈合速度降低相关。虽然小鼠中MRP8表达的调控可能与人类皮肤不同,但我们认为MRP8和MRP14的升高表达可能对伤口愈合中的炎症具有相关治疗作用。