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目的:探讨p63基因编码的蛋白亚型△Np63α在创面愈合表皮再生中的表达及其生物学意义。方法:①在3周龄小鼠背部用直径1.6cm的环钻压切制备出一个2.0cm~2全层皮肤缺损的圆形创面,并将切除的皮肤原位缝合以覆盖创面。②将18只昆明小鼠随机分成两组,实验组12只小鼠均在背部用环钻制备圆形创面;而对照组6只小鼠不致伤。分别于1d、2d、3d、7d、14d、21d在实验组创缘和对照组小鼠背部相同部位切取全层皮肤组织标本,用抗鼠多克隆抗体P40进行抗△Np63免疫组化染色。结果:△Np63α在正常皮肤中的特征性表达模式是强阳性染色限于生发层的角质形成细胞核。伤后1~2d,表皮开始迁移,与对照组正常皮肤相比,迁移表皮舌基底层细胞△Np63α表达下调。伤后3d,△Np63α有较强的表达出现在创缘和迁移的表皮舌中,阳性染色限于基底层和其上方1~2层的角质形成细胞,而创面尚未被上皮覆盖处则没有阳性细胞。伤后5~7d,△Np63a有更强的表达出现在创缘和迁移表皮舌的基底层角质形成细胞。伤后14~21d创面愈合,△Np63α在覆盖创伤区表皮的基底层和棘层的细胞中高表达,并且其表达在表皮中长期和持续的存在。结论:△Np63α可能通过调控表皮修复中的细胞增殖和细胞迁移而参与正常皮肤的创面愈合过程。
Objective: To investigate the expression of p63 gene-encoded protein Np63α in wound healing epidermis and its biological significance. Methods: ① A circular wound of 2.0 cm ~ 2 full thickness skin defect was prepared on the back of a 3-week-old mouse with a diameter of 1.6 cm and the excised skin was sutured in situ to cover the wound. ② 18 Kunming mice were randomly divided into two groups. Twelve mice in the experimental group were prepared circular wound on the back with trephine; while six mice in the control group were not injured. Full-thickness skin specimens were cut at the same site on the back of the experimental group and the control group on day 1, day 2, day 3, day 7, day 14, and day 21 respectively. The anti-△ Np63 immunohistochemical staining was performed using the anti-mouse polyclonal antibody P40. Results: The characteristic expression pattern of △ Np63α in normal skin is that the strong positive staining is limited to the keratinocyte nucleus of the germinal layer. 1 ~ 2 days after injury, the epidermis began to migrate. Compared with the normal skin of the control group, the expression of △ Np63α was down-regulated in the basal lamina of migrating epidermis. On the 3rd day after injury, △ Np63α had a stronger expression in the tongue of the epidermis and the migrating epidermis. The positive staining was limited to the basal layer and the keratinocytes above the basal layer and the layer above it, while the wound surface was not covered by the epithelium and had no positive cells . From 5 to 7 days after injury, △ Np63a had a stronger expression in basal layer of keratinocytes that located on the wound edge and migrate the epidermis tongue. The wound healed 14 to 21 days after injury, and △ Np63α was highly expressed in the basal layer and the stratum spinous layer covering the epidermis of the wound area, and the expression was persistent in the epidermis for a long time. Conclusion: △ Np63α may participate in the wound healing process of normal skin by regulating cell proliferation and cell migration during epidermal repair.