小鼠皮肤切创愈合过程中iNOS和eNOS表达的免疫组织化学研究

目的观察小鼠皮肤切创愈合过程中,iNOS和eNOS在损伤区及损伤周边区内的表达及其变化规律。方法小鼠背部制作全层切创,应用免疫组织化学技术观察伤后切创组织中iNOS和eNOS的表达,并和无切创的小鼠作为对照。结果损伤区及损伤周边区细胞,伤后3h的损伤皮肤组织中可见少量的多型核细胞表达iNOS和eNOS,伤后6～24h,大部分浸润的中性粒细胞和单核细胞为iNOS和eNOS阳性。随着时间的延长,iNOS和eNOS阳性细胞以单核细胞和成纤维细胞为主。伤后3hiNOS的阳性细胞比率较低,6h～1d持续性增加并于1d达到最高峰,3～7d维持在一个相对稳定的水平直至伤后10d再次达高峰,10～14d开始下降。eNOS的阳性细胞率在伤后1～3h表达较低,6h～3d持续性增高,并在3d达到最高峰,在其后的5d内保持稳定表达,之后开始下降。而且损伤区及损伤周边区、特别是肉芽组织内的新生血管可见iNOS和eNOS不同强度的表达。结论小鼠皮肤切创愈合过程中,iNOS和eNOS在损伤周边区内多型核细胞、单核细胞和成纤维细胞中表达,其时序性变化可望用于皮肤损伤时间的推断。

Immunohistochemical study on expression of iNOS and eNOS during skin incised wound healing in mice

] Objective To investigate the expressions of iNOS and eNOS during skin incised wound healing and theapplicability of time-dependent expressions of iNOS and eNOS to timing of wound. Methods An incised woundwas made in the dorsal skin of mouse . By using S-P method, the expression of iNOS and eNOS were studied in cu-taneous incised wound as well as normal skin in control mice. Results iNOS and eNOS expressed in epidermis,hair follicle, sebaceous gland, and the eNOS were also found in vascular endothelium in control group. Three hoursafter injuryed, the expression of iNOS and eNOS were detected in polymorphonuclear cells (PMNs) in the woundand peripheral region. From 6h to 24h of wound interval, the expression of NOS isoforms were identified in a largenumber of infiltrating PMNs and part of mononuclear cells (MNCs), afterwards the MNCs and fibroblastic cells(FBCs) accounted for the most part of the iNOS and eNOS positive cells. In addition, during wound healing, the ex-pressions of iNOS and eNOS were noticed in different level in vascular epithelial cells, especially neonatal capil-lary vessel in granulation tissue. Conclusion The results suggest that iNOS and eNOS express in polymorphonu-clear cells, macrophages and fibroblasts during healing process of skin incised wound in mice. Furthermore, iNOSand eNOS may play pivotal roles in all the events relating to wound healing, and the time-dependent expressions ofiNOS and eNOS may possibly be used as newmarkers for the wound age determination.