皮肤创伤愈合过程中TGF-β_1mRNA表达与损伤时间关系的实验研究.docVIP

皮肤创伤愈合过程中TGF-β1mRNA表达与损伤时间关系的实验研究 作者：李学锋1；王慧君1；罗红2　　(1南方医科大学南方医院病理科，广东??广州??510515；2中山大学，广东??广州??510215) 摘要：目的??研究TGF-β1 mRNA在皮肤创伤愈合中的表达变化规律及其在损伤时间推断中的价值。方法 用实时PCR技术检测不同造创时间(生前伤15 min~2 周，死后伤1~6 h)小鼠皮肤切创创缘组织TGF-β1 mRNA含量，并同时检测GAPDH含量对结果进行均一化。结果??TGF-β1 mRNA于伤后24 h显著增高(和对照组比较P<0.01)，48 h和72 h时还保持在较高水平。死后伤1 h和3 h时，TGF-β1 mRNA的表达有增高的趋势，6 h则降至对照水平。结论??TGF-β1 mRNA在小鼠皮肤创伤后有明显的规律性表达，能作为一种代表性细胞因子参与有一定存活时间(6 h以上)的生前伤损伤时间的推断。实时PCR技术能对mRNA的表达水平进行较为精确的检测，是目前损伤时间推断研究中用于细胞因子定性和定量研究的有效方法。 关键词：TGF-β1；创伤修复；损伤时间推断；实时PCR 中图分类号：R338??文献标识码：A??文章编号：1673-4254(2006)01-0059-03 Temporal relation of transforming growth factor-β1 mRNA expression with injury in the healing process of mouse skin woundsLI Xue-feng1；WANG Hui-jun1；LUO Hong21Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; 2Sun Yat-sen University, Guangzhou 510215, ChinaAbstract: Objective To explore the time-related expression of transforming growth factor (TGF)-β1 mRNA in the healing process of mouse skin wounds and its potential application in forensic wound age estimation. Methods Real-time PCR was used to quantify TGF-β1 mRNA expression in mouse skin wounds induced at different time points (15 min to 2 weeks before and 1 to 6 h after death). The expression of GAPDH was also detected for normalization of TGF-β1 mRNA level. Results TGF-β1 mRNA in the antemortem wounds was elevated significantly 24 h after the injury (P<0.01 by Dunnet test) and maintained the high level at 48 and 72 h. In postmortem wounds, TGF-β1 mRNA tended to increase 1 and 3 h after injury, and decreased to the control level at 6 h. Conclusions The expression of TGF-β1 mRNA is closely related to the time of wound and can therefore be used as a representative cytokine marker for dating the skin wounds induced over 6 h before death. Real-time PCR for detecting dynamic expression of TGF-β1 mRNA in wound healing process provides a reliable means for qualitative and quantitative cytokine analysis for forensic