小鼠皮肤切创愈合过程中p-JNK变化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JNK（p-JNK）的变化及不同损伤时间p-JNK的变化规律。方法 应用免疫组织化学和Westernblot方法检测小鼠皮肤切创后各个时间段p-JNK的变化情况。结果 正常组织中有少量p-JNK阳性着色。伤后3～12h损伤区p-JNK阳性着色主要在中性粒细胞,1～5dp-JNK阳性着色主要为单核细胞和成纤维细胞,7～14dp-JNK阳性着色以成纤维细胞为主。阳性细胞率3h～1d逐渐增高,1d达到峰值,3～5d有所下降,7d阳性细胞率达第二峰值。10～14d逐渐降低。Westernblot显示各个时间段均有p-JNK的阳性条带,其中12h和3d为p-JNK含量的2个高峰。结论 小鼠皮肤切创愈合过程中p-JNK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非常重要的作用。同时P-JNK的规律性变化可用于损伤时间推断。

The time-dependent changes of phospho-JNK expression during the skin incised wound healing in mice

Objective To investigate the changes of phospho-JNK(p-JNK) during the incised wound healing of the skin in mice and to explore the rule of the time-dependent change of p-JNK in wound age determination.Methods The changes of p-JNK expression in incised skin wound were detected by immunohistochemistry and Western blot.Results There was a minimal baseline staining of p-JNK in control mouse skin.Changes of p-JNK expression were mainly detectable in neutrophils in the wound specimens from 3 hours to 12 hours after injury.Afterwards,the p-JNK positive cells were mostly mononuclear cells and fibroblasts between post-injury day 1 and day 5,whereas the p-JNK positive cells were mostly fibroblasts between post-injury day 7 and day 14.Morphometrically,the ratio of the p-JNK positive cells to the total increased gradually in the wound specimens from 3 hours to day 1,and maximized at day 1 with a slight decrease from post-injury day 3 to day 5.The ratio showed a second peak in the specimens of day 7,and then decreased gradually from post-injury day 10 to day 14.The changes of p-JNK expression were 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 p-JNK may play a pivotal role in inducing apoptosis of neutrophils,mononuclear cells,and fibroblasts during skin wound healing and meanwhile,p-JNK may be a potentially useful marker for wound age determination.