骨桥蛋白在新生大鼠背部皮肤创伤愈合过程中的表达

Expression and Distribution of Osteopontin in Cutaneous Wound Healing

目的:检测骨桥蛋白(osteopontin,OPN)在新生大鼠皮肤及创伤愈合过程中的表达,探讨OPN参与皮肤创伤愈合的可能机制。方法:在7 d龄SD大鼠背部皮肤做直径为1 cm的圆形全层皮肤切口,运用免疫组织化学方法检测OPN阳性细胞在皮肤及创伤愈合过程中的分布情况,并分别运用RT-PCR及Western blot检测OPN在创伤愈合不同时间点(1、3、5及7 d)的表达情况。结果:在大鼠正常皮肤,免疫组织化学结果显示OPN阳性细胞分布在毛囊、皮脂腺及汗腺,表皮则无表达;RT-PCR及Western blot均未检测到OPN在正常皮肤中的表达。从创伤后第1天起,OPN阳性细胞开始集中分布在创伤边缘的真皮部位,且在第3天达到高峰,第5天表达开始减弱。RT-PCR及Western blot结果一致,OPN相对表达量在第3天达到高峰。结论:研究结果表明OPN参与了皮肤创伤愈合,尤其在创伤修复的炎症期及细胞外基质沉积期。

Objective： The aim of this study was to investigate the localization of osteopontin （OPN） in normal rat skin as well as its expression variation during the process of wound healing. Methods： Circular full-thickness skin wound （lcm diameter） was made in the dorsal aspect of Sprague-Dawley （SD） rat. After 1, 3, 5 and 7 days, skin wound sample were collected and examined by immunohistochemistry, reverse-transcription polymerase chain reaction （RT-PCR） and immunoblotting. Results： In normal skin, positive expression of OPN only revealed in the hair follicles, sebaceous glands and sweat glands, in the cutaneous tissue proper, OPN expression appeared to be negative. The expression of OPN was undetected in SD rat skin by RT-PCR and Western blot techniques. However during the course of cutaneous wound healing, OPN expression showed evident, which peaked at the third day. Conclusion： The results of this study suggest that OPN may participate in skin wound healing, particularly in inflammation and proliferation phases, as a chemotactic cytokine.