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

研究伤口愈合过程中细胞因子 mRNA 表达以确定伤口年龄。

基本信息

批准号：
12670402
负责人：
AOKI Yasuhiro
金额：
$ 1.73万
依托单位：
Iwate Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670402/
关键词：
injury wound aging wound healing bFGF VEGF fibronectin quantitative PCR in situ hybridization サイトカイン遺伝子発現 fibronectin 免疫組織化学損傷創傷治癒法医学

项目摘要

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.

本研究通过动物实验研究了细胞因子mRNA表达作为创伤老化标志物的可行性。在5 ~ 6周龄雄性DDY小鼠背部皮肤上切出2cm全厚切口，切口后0.5、1、3、8、24、72、144或240小时处死。用特异性抗体免疫组化法检测碱性成纤维细胞生长因子（bFGF）和血管内皮生长因子（VEGF）的表达。对于伤口年龄的测定，bFGF，这是在早期0.5- 1小时阶段和晚期24- 144小时阶段检测到的，似乎是上级VEGF，因为它的早期表达，因为它更持久的表达在真皮成纤维细胞增加死后间隔。因此，随后使用定量PCR和原位杂交技术分析伤口愈合过程中bFGF mRNA的表达。除皮肤创伤外，还检测了皮肤、脑、肝和肾组织的mRNA动态。bFGF mRNA在损伤皮肤中的表达在1小时达到高峰，在表皮细胞、成纤维细胞、内皮细胞和中性粒细胞中检测到。脑损伤后1小时开始升高，48小时达高峰。星形胶质细胞是vFGFmRNA的主要来源。然而，在肝和肾损伤中，bFGF mRNA表达在24小时时在内皮细胞、肝的中性粒细胞和肾的肾小球中略有增加。这些结果表明，来自外胚层的皮肤和脑的mRNA动态与肝脏和肾脏的mRNA动态有很大不同。换言之，bFGF mRNA的时间依赖性表达可作为外胚层起源器官的有用的创伤老化标志物。