Zhu Jian-Wu, Lai Xi-Nan, Wanc Zheng-Guo, Huang Hui, Wang Li-Li, Ning Jia-Lin
Institute of Field Surgery, Daping Hospital, Third Military Medical University, Chongqing 400042, PR China.
Zhonghua Shao Shang Za Zhi. 2007 Jun;23(3):204-7.
To investigate the effect of substance P (SP) on the migration and differentiation of epidermal stem cells (ESCs) to hair follicle, and its mechanism.
ESCs were cultured in vitro, and confirmed by positive staining of K19 and integrin beta1 with immunohistochemistry. SP was added into the culture of ESCs which were labelled with 5-BrdU, and the cell cultures were divided into control, 10(-5) mol/L SP, 10(-6) mol/L SP, and 10(-7) mol/L SP groups according to the different doses of SP addition. Cell suspension (0.3 ml) containing SP was injected into the dermis in the back of nude mice. Repeated injection of the equal amount of cell suspension in the same place was carried out on 4, 7, 10 and 14 days after first injection. The cells in control group received the same treatment but without SP. The skin specimens in the area of cell culture injection and the normal skin remote from cell injection were harvested for the histological examination and hair follicle counting by immunohistochemistry and electronmicroscope 28 days after injections.
Hair follicles in scattered distribution were observed in 10(-5) mol/L SP group,but some of them were defective in development. Hypoplasic hair follicle and a few hair follicles with distinct structure were observed in 10(-5) mol/L SP group. Large amounts of hair follicles with distinct structure in deep dermis and subcutaneous tissue were observed in 10(-6) mol/L SP, 10(-7) mol/L SP groups, and some of them showed positive staining of brown BrdU in the hair root, and most of them showed positive staining of brown beta-catenin, but a few of them showed developmental defect. In contrast, hypoplasia of hair follicle underneath epidermis and deep layer of dermis with positive staining of brown BrdU and beta-catenin in epidermis were observed in control group. The number of hair follicles in 10(-6)mol/L SP, 10(-7) mol/L SP groups [(1.9 +/- 1.2 ), (1.3 +/- 0.8)] was obviously less than that in control group [(10. 5 +/- 1.2), P < 0.01].
SP can induce ESCs to migrate from the basal layer into hair follicle, and this effect is dependent on the SP concentration. SP can also elevate the expression of beta-catenin in ESCs,which induces its differentiation to hair follicles.
探讨P物质（SP）对表皮干细胞（ESCs）向毛囊迁移和分化的影响及其机制。
体外培养ESCs，通过免疫组化K19和整合素β1阳性染色进行鉴定。将5-溴脱氧尿嘧啶核苷（5-BrdU）标记的ESCs培养物中加入SP，根据SP添加剂量不同将细胞培养物分为对照组、10⁻⁵mol/L SP组、10⁻⁶mol/L SP组和10⁻⁷mol/L SP组。将含SP的细胞悬液（0.3 ml）注射到裸鼠背部真皮内。首次注射后第4、7、10和14天在同一部位重复注射等量细胞悬液。对照组细胞接受相同处理但不添加SP。注射28天后，取细胞培养注射部位及远离细胞注射的正常皮肤标本，通过免疫组化和电子显微镜进行组织学检查和毛囊计数。
10⁻⁵mol/L SP组可见散在分布的毛囊，但部分毛囊发育不良。10⁻⁶mol/L SP组可见毛囊发育不全，少数毛囊结构清晰。10⁻⁶mol/L SP组、10⁻⁷mol/L SP组在真皮深层和皮下组织可见大量结构清晰的毛囊，部分毛囊毛根呈棕色BrdU阳性染色，多数毛囊β-连环蛋白呈棕色阳性染色，但少数毛囊有发育缺陷。相比之下，对照组表皮下和真皮深层毛囊发育不全，表皮有棕色BrdU和β-连环蛋白阳性染色。10⁻⁶mol/L SP组、10⁻⁷mol/L SP组毛囊数[（1.9±1.2），（1.3±0.8）]明显少于对照组[（10.5±morphology was observed in 10(-6) mol/L SP group, and a few hair follicles with distinct structure were observed in 10(-5) mol/L SP group. Large amounts of hair follicles with distinct structure in deep dermis and subcutaneous tissue were observed in 10(-6) mol/L SP, 10(-7) mol/L SP groups, and some of them showed positive staining of brown BrdU in the hair root, and most of them showed positive staining of brown beta-catenin, but a few of them showed developmental defect. In contrast, hypoplasia of hair follicle underneath epidermis and deep layer of dermis with positive staining of brown BrdU and beta-catenin in epidermis were observed in control group. The number of hair follicles in 10(-6)mol/L SP, 10(-7) mol/L SP groups [(1.9 +/- 1.2 ), (1.3 +/- 0.8)] was obviously less than that in control group [(10. 5 +/- 1.2), P < 0.01].
SP can induce ESCs to migrate from the basal layer into hair follicle, and this effect is dependent on the SP concentration. SP can also elevate the expression of beta-catenin in ESCs,which induces its differentiation to hair follicles.1.2），P<0.01]。
SP可诱导ESCs从基底层迁移至毛囊，且该作用依赖于SP浓度。SP还可提高ESCs中β-连环蛋白的表达，诱导其向毛囊分化。
