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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。
OBJECTIVE: To observe the expression of iNOS and eNOS in the wound healing and peri-injured areas during incised wound healing in mice. Methods Full thickness incision was made on the back of mice. Immunohistochemistry was used to observe the expression of iNOS and eNOS in incisional wounds. The mice were used as controls. RESULTS: A small amount of iNOS and eNOS were seen in damaged and damaged peripheral skin cells at 3 h after injury. Most of the infiltrated neutrophils and monocytes expressed iNOS and eNOS at 6 to 24 h after injury, eNOS positive. Over time, iNOS and eNOS positive cells to monocytes and fibroblasts. The ratio of positive cells in 3hiNOS after injury was low, increased continuously from 6h to 1d and reached its peak at 1d, maintained at a relatively stable level from 3 to 7d and peaked again at 10d, and then decreased from 10 to 14d. The positive cell rate of eNOS was lower at 1 ~ 3h after injury and persistently increased at 6h ~ 3d, and reached the peak at 3d, then remained stable within 5d and then began to decrease. Moreover, the expression of iNOS and eNOS with different intensity was observed in the injured area and the injured peripheral area, especially the neovascularization in granulation tissue. Conclusions During incised wound healing, iNOS and eNOS are expressed in the peripheral blood of polymorphonuclear cells, monocytes and fibroblasts. The time-dependent changes of iNOS and eNOS are expected to be used to infer the time of skin injury.