小鼠皮肤切创愈合过程中磷酸化 p38MAPK 表达及其与时间的相关性

目的探讨小鼠皮肤切创愈合过程中损伤区磷酸化p38MAPK(p-p38MAPK)的表达情况,以及不同损伤时间p-p38MAPK的变化规律。方法应用免疫组织化学和Westernblot的方法检测33例小鼠皮肤切创后各个时间段p-p38MAPK表达情况。结果正常组织中有少量的p-p38MAPK表达。伤后3～12h损伤区p-p38MAPK主要表达于中性粒细胞中,1～5dp-p38MAPK阳性表达主要为单核细胞和成纤维细胞。7～14dp-p38MAPK阳性表达主要以成纤维细胞为主。阳性细胞率3～12h逐渐升高,1d有所下降,3、5d阳性细胞率保持在高水平,7～14d阳性细胞率逐渐降低。Westernblot显示各个时间段均有p-p38MAPK的表达。其中12h和3d为2个p-p38MAPK含量的高峰。结论在小鼠切创愈合过程中p-p38MAPK在损伤区对诱导中性粒细胞、单核细胞和成纤维细胞的凋亡起重要的作用,同时p-p38MAPK的规律性表达可用于损伤时间的推断。

A study on the time-dependent expression of phospho-p38MAPK during the skin incised wound healing in mice

Objective To investigate the expressions of phospho-p38MAPK(p-p38MAPK) during the incised wound healing of the skin in mice and to explore the applicability of the time-dependent expressions of p-p38MAPK in the determination of wound age. Methods The expression of p-p38MAPK in incised skin wound was detected by immunohistochemistry and Western blot. Results There was a minimal baseline expression of p-p38MAPK in control mouse skin. Expression of p-p38MAPK was mainly detectable in neutrophils in the wound specimens from 3 hours to 12 hours after injury. Afterwards,the p-p38MAPK positive cells were mostly mononuclear cells and fibroblasts between post-injury day 1 and day 5,whereas the p-p38MAPK positive cells were mostly fibroblasts between post-injury day 7 and day 14. Morphometrically,the ratio of the p-p38MAPK positive cells to total number of the cells increased gradually in the wound specimens aged from 3 hours to 12 hours,and maximized in the wound specimens aged 12 hours with a slight decrease at 24 hours after injury. The ratio maintained at high level from post-injury day 3 to day 5,and then decreased gradually from post-injury day 7 to day 14. The expression of p-p38MAPK was observed throughout the wound healing stages by Western blot as well with a peak expression occurring between 12 hour and day 3 after injury. Conclusion Our data suggest that p-p38MAPK may play an important role in inducing apoptosis of neutrophils,mononuclear cells,and fibroblasts during skin wound healing and meanwhile,p-p38MAPK may be a potentially useful marker for wound age determination.