皮肤创伤愈合过程中TGF-β_1 mRNA表达与损伤时间关系的实验研究

Temporal relation of transforming growth factor-β_1 mRNA expression with injury in the healing process of mouse skin wounds

目的研究TGF-β1mRNA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-β1mRNA含量,并同时检测 GAPDH含量对结果进行均一化。结果TGF-β1mRNA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-β1mRNA的表达有增高的趋势,6 h则降至对照水平。结论TGF-β1mRNA 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(6h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。

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 mainrained 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 wound age estimation.