Studies on mRNA Expressions of Cytokines during Wound Healing for Determination of Wound Age.

为探讨细胞因子基因表达作为创伤老化标志物的可行性，进行了动物实验。5~6周龄雄性DDY小鼠背部皮肤作2 cm全厚切口，分别于术后0.5、1、3、8、24、72、144、240h处死。免疫组织化学方法检测碱性成纤维细胞生长因子和血管内皮生长因子的表达。在伤后0.5~1小时和24~144小时的早期阶段检测到的碱性成纤维细胞生长因子似乎优于血管内皮生长因子，因为它的早期表达和在真皮成纤维细胞中的持续表达随着死后时间的延长而延长。因此，随后利用定量聚合酶链式反应和原位杂交技术分析了伤口愈合过程中碱性成纤维细胞生长因子基因的表达。除真皮创面外，还观察了真皮、脑、肝、肾等组织的mRNA动态变化。伤后1h皮肤组织中bFGFm RNA表达达高峰，在表皮细胞、成纤维细胞、内皮细胞和中性粒细胞中均有表达。损伤大脑1h开始升高，48h达高峰。星形胶质细胞是损伤脑内vFGFm RNA供应的主要来源。然而，在肝和肾损伤中，在24小时内内皮细胞、肝脏的中性粒细胞和肾脏的肾小球中bFGFmRNA的表达略有增加。这些结果表明，外胚层来源的皮肤和脑与肝脏和肾脏的mRNA动态有很大的不同。也就是说，bFGFm RNA的时间依赖性表达可能是外胚层起源器官的有用的创伤老化标志物。

An animal experimental study was carried out to investigate availability of mRNA expressions of cytekines as markers for wound aging. A 2 cm full thickness incision was made on the dorsal skin of male DDY mice of 5 to 6 weeks old, and they were killed 0.5, 1, 3, 8, 24, 72, 144, or 240 hours after the incision. The expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) were examined immunohistochemically using specific antibodies. For wound age determination, bFGF, which was detected in the early 0.5- to 1-hour phases and the late 24- to 144-hour phases, seemed to be superior to VEGF because of its earlier expression and because of its more persistent expression in dermal fibroblasts with increasing postomortem interval. Therefore, expressions of bFGF mRNA during wound healing were subsequently analyzed using quantitative PCR and in situ hybridization techniques. Beside the dermal wound, mRNA dynamics of dermal, cerebral, hepatic and renal ones were also examined. bFGF mRNA expressions in the injured skin peaked at 1 hour, and they were detected in epidermal cells, fibroblasts, endothelial cells and neutrophils. Those of the injured cerebrum increased from 1 hour and peaked at 48 hours. The astrocytes were main source of vFGF mRNA supply in the injured brain. In hepatic and renal injuries, however, bFGF mRNA expressions increased slightly at 24 hours in endothelial cells, in neutrophils of the liver, and in glomeruli of the kidney. These results suggested that the mRNA dynamics of skin and brain derived from ectoderm were quite different from those of liver and kidney. In other words, time-dependent expressions of bFGF mRNA could be useful wound aging marker for the organs of ectodermal origin.