小鼠皮肤切创黏着斑激酶和磷酸化黏着斑激酶的表达及其变化规律

目的研究小鼠皮肤切创愈合过程中,黏着斑激酶（focal adhesion kinase,FAK）和磷酸化黏着斑激酶（phospho—FAK,p—FAK）在损伤区及损伤周边区内的表达及其变化规律。方法应用免疫组织化学染色技术和免疫印迹（Western blot）方法观察小鼠背部切创后损伤区及损伤周边区FAK及p-FAK的表达情况。结果伤后3h,FAK和p-FAK主要表达于多核粒细胞;伤后6～24h,主要表达于大部分浸润的多核粒细胞和单核细胞;伤后3～14d。主要表达于单核细胞和成纤维细胞。伤后FAK的阳性细胞率逐渐增高．于3d达到高峰．随后迅速下降;伤后p-FAK的阳性细胞率也逐渐增加,于12h达到高峰,之后逐渐下降。Westernblot结果显示,正常皮肤中FAK及P—FAK均有表达。FAK表达强度变化不是十分明显,但仍可见在3d时含量达到最高。p-FAK蛋白含量伤后逐渐增加．12h表达最强,随后逐渐下降。结论在小鼠皮肤切创愈合过程中,FAK和p-FAK阳性细胞率呈时间规律性变化．可用于皮肤切创损伤时间的推断。p-FAK呈现出明显的时间规律性变化,说明作为皮肤损伤时间的推断指标要优于FAK。

Expressions and time-dependent changes of FAK and phospho-FAK during skin incised wound healing in mouse

Objective To study the expressions of focal adhesion kinase(FAK) and phospho-FAK(p-FAK) during skin incised wound healing and the applicability of time-dependent expressions of FAK and p-FAK.Methods The expression of FAK and p-FAK in cutaneous incised wound in mouse were investigated by immunohistochmeistry and Western blotting.Results FAK and p-FAK expression were detected in polymorphonuclear cells(PMNs) in the wound and adjacent regions 3 hours post-injury.The expressions of FAK and p-FAK were detected in a large number of infiltrating PMNs and some of mononuclear cells(MNCs) from 6 to 24 hours after injury.The MNCs and fibroblastic cells(FBCs) accounted for most part of the FAK and p-FAK positive cells from 3 to 14 days after injury.The numbers of FAK-positive cells increased continuously,reaching a peak at post-injury day 3,and then started to decrease from post-injury day 5 to 14.The changes of p-FAK-positive cells were similar to that of the FAKs,and reached a peak at 12 hours after injury.Conclusion Both FAK and p-FAK displayed a time-dependent expression during skin incised wound healing in mouse,with p-FAK being superior to FAK.Both FAK and p-FAK may potentially be used as new markers for determination of the wound interval.