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目的探讨小鼠皮肤切创愈合过程中损伤区M3受体的表达及其随时间的变化规律。方法应用免疫组织化学染色和Western印迹技术检测小鼠皮肤切创后不同时间段M3受体的变化情况。结果正常皮肤组织的表皮、毛囊、皮脂腺、汗腺、皮肌层等均表达M3受体。伤后6～12h,损伤区M3受体阳性细胞以多形核粒细胞为主;1～3d,M3受体阳性细胞以单个核细胞和成纤维细胞为主;5～14d,M3受体阳性细胞主要为成纤维细胞。阳性细胞率在伤后6～12h逐渐增高,12h达高峰,1～5d略有下降,但保持较高水平,7d阳性细胞率再次达到峰值,随后逐渐降低。通过Western印迹法检测显示,各个时间段均有M3受体阳性条带,其中12 h和7d为M3受体两个表达高峰。结论多形核粒细胞、单个核细胞和成纤维细胞均表达M3受体,提示其可能在皮肤切创愈合过程中起重要的作用;M3受体表达的规律性变化可用于损伤时间的推断。
Objective To investigate the expression of M3 receptor in the lesion area during incised wound healing in mice and its change with time. Methods Immunohistochemical staining and Western blotting were used to detect the changes of M3 receptors at different time points after skin incision in mice. Results The normal skin tissue of the epidermis, hair follicles, sebaceous glands, sweat glands, muscularis and so on all express M3 receptor. From 6 to 12 hours after injury, the majority of M3 receptor positive cells in the injured area were polymorphonuclear neutrophils. In the first to third days, the majority of M3 receptor positive cells were mononuclear cells and fibroblasts. From 5 to 14 days, M3 receptor positive The cells are mainly fibroblasts. The rate of positive cells increased gradually from 6 to 12 hours after injury, reached the peak at 12 hours and decreased slightly from 1 to 5 days. However, the rate of positive cells peaked again on the 7th day and then decreased gradually. Western blotting showed that there were M3 receptor positive bands in each time period, of which 12 h and 7 d were the peak expression of M3 receptor. Conclusion All of the polymorphonuclear neutrophils, mononuclear cells and fibroblasts express M3 receptor, suggesting that they may play an important role in incised wound healing. The regular change of M3 receptor expression can be used to infer the damage time.