慢性心理应激导致皮肤氧化应激损伤及毛囊生长期延迟

发布时间：2018-09-12 12:14

【摘要】：目的：1、建立慢性束缚应激（Chronic restraint stress，CRS）模型，观察心理应激对小鼠毛囊周期的影响。2、研究P物质（substance P, SP），氧化应激（Oxidativestress），肥大细胞（Mast cell，MC）在心理应激引起毛囊周期改变这一过程中的作用。3、探讨脱发的发病机制，为其预防、治疗及新药开发提供新思路和更可靠的实验依据。方法：1、采用C57BL/6雄鼠建立CRS模型，与Control组比较，从肉眼和组织形态学观察动物毛囊周期的改变。2、在原有Control，CRS两组基础上，加入腹腔注射抗氧化剂和SP受体阻断剂两组，，所有小鼠随机分为四组：Control组、CRS组、CRS+抗氧化剂组（Tempol组）、CRS+SP受体阻断剂组（RP67580组）。使用脱毛术诱导小鼠背部毛囊进入生长期，选取脱毛后第9天和第19天作为时间点采集标本，观察体重、行为、皮肤颜色和毛囊组织形态的改变并对毛囊进行评分，检测血浆皮质醇水平，皮肤组织中硫代巴比妥酸反应产物（TBARS）水平，超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）活力；甲苯胺蓝染色切片观察皮肤肥大细胞脱颗粒比例；免疫荧光法观察皮肤SP免疫反应阳性（SP+）神经纤维数量。结果：1、脱毛后第9天，与Control组相比，CRS组皮肤呈粉色，灰度率显著升高（P0.05），毛囊平均分显著降低（P0.05），生长期延迟出现；脱毛后第19天，与Control组相比，CRS组皮肤皮肤呈黑色，灰度率和毛囊平均分均显著降低（P0.05），退行期延迟出现。2、（1）与Control组相比，CRS组、Tempol组、RP67580组小鼠体重和旷场实验中3min内运动距离均显著降低（P0.05），血浆皮质酮水平显著升高（P0.05），提示慢性束缚应激模型造模成功；（2）与Control组相比，CRS组皮肤组织TBARS水平显著升高（P0.05），SOD、GSH-Px活力显著降低（P0.05）；同CRS组相比，Tempol组和RP67580组皮肤TBARS水平显著降低（P 0.05）、GSH-Px活力显著升高（P0.05）；（3）脱毛后第9天，与CRS组相比，Tempol组和RP67580组皮肤呈灰色，灰度率显著降低（P0.05），毛囊平均分显著升高（P0.05）；脱毛后第19天，与CRS组相比，Tempol组和RP67580组皮肤呈灰色，灰度率和毛囊平均分显著升高（P0.05），毛囊周期未见明显延迟；（4）与Control组相比，CRS组皮肤SP+神经纤维数量明显升高（P0.05）；与CRS组相比，Tempol组和RP67580组皮肤SP+神经纤维数量明显降低（P0.05）。（5）同C组相比，CRS组皮肤肥大细胞脱颗粒比例明显升高（P0.05）；与CRS组相比，Tempol组和RP67580组皮肤肥大细胞脱颗粒比例明显降低（P0.05）。结论：1、慢性束缚应激可以导致毛囊周期的异常，表现为生长期延迟出现。2、慢性束缚应激可以引起皮肤组织氧化应激损伤。3、慢性束缚应激可以激活SP-MC通路，导致SP+神经纤维数量上升，肥大细胞脱颗粒。4、在皮肤组织中氧化应激与SP-MC通路之间存在交互作用。5、抗氧化剂和SP受体阻断剂可以部分的改善慢性束缚应激对毛囊的负性作用。6、慢性束缚应激是研究心理应激与毛囊之间关系的理想模型。
[Abstract]:Objective to establish (Chronic restraint stress,CRS model of chronic restraint stress. To observe the effect of psychological stress on hair follicle cycle in mice, to study the role of substance P (substance P, SP), oxidative stress (Oxidativestress), mast cells (Mast cell,MC) in the process of hair follicle cycle change induced by psychological stress, to explore the pathogenesis of alopecia and to prevent it. Treatment and new drug development provide new ideas and more reliable experimental basis. Methods C57BL/6 male rats were used to establish CRS model. Compared with Control group, the changes of hair follicle cycle were observed by naked eye and histomorphology. On the basis of the original Control,CRS group, the rats were given intraperitoneal injection of antioxidants and SP receptor blockers. All mice were randomly divided into four groups: CRS antioxidant group (Tempol group) and CRS SP receptor blocker group (RP67580 group). The hair follicles of mice were induced to enter the growth stage by depilation. The specimens were collected on the 9th and 19th day after hair removal. The changes of body weight, behavior, skin color and hair follicle tissue morphology were observed and the hair follicles were scored. Plasma cortisol level, thiobarbituric acid reaction product (TBARS) level, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activity and toluidine blue staining section were detected. The number of SP immunoreactive (SP) nerve fibers in skin was observed by immunofluorescence method. Results on the 9th day after hair removal, the skin of Control group was pink, the gray scale was significantly higher (P0.05), the average score of hair follicle was significantly lower (P0.05), and the growth period was delayed, and on the 19th day after hair removal, the skin of Control group was black compared with that of Control group. The gray level and the average score of hair follicles were significantly decreased (P0.05), and the delay of receding phase appeared 0.2. (1) compared with the Control group, the body weight and the distance of movement in 3min in the Tempol group were significantly lower than those in the Control group (P0.05), and the plasma corticosterone level was significantly increased (P0.05). The model of chronic restraint stress was established successfully. (2) compared with Control group, the level of TBARS in skin tissue was significantly increased (P0.05), the activity of GSH-Px in skin was significantly decreased (P0.05), the level of GSH-Px in skin of CRS group and RP67580 group was significantly lower than that of CRS group (P 0.05), and the activity of GSH-Px in skin tissue was significantly increased (P0.05) on the 9th day after depilation (P0.05), and that in RP67580 group was significantly lower than that in CRS group (P0.05). Compared with CRS group, the skin of Tempol group and RP67580 group were gray, gray scale was significantly decreased (P0.05), the average score of hair follicle was significantly increased (P0.05), the skin of Tempol group and RP67580 group was gray compared with CRS group on the 19th day after hair removal. The gray scale and the average score of hair follicles were significantly increased (P0.05), but the hair follicle cycle was not significantly delayed. (4) compared with Control group, the number of SP nerve fibers in skin was significantly increased (P0.05). Compared with CRS group and RP67580 group, the number of skin SP nerve fibers decreased significantly (P0.05). (5). Compared with C group, the percentage of mast cell degranulation increased significantly (P0.05); compared with CRS group and RP67580 group, the degranulation ratio of skin mast cells decreased significantly (P0.05). Conclusion: 1. Chronic restraint stress can lead to abnormal hair follicle cycle, which is characterized by delayed appearance of .2in growth period. Chronic restraint stress can induce oxidative stress damage of skin tissue. Chronic restraint stress can activate SP-MC pathway. Leading to an increase in the number of SP nerve fibers, Mast cell degranulation. 4, there is interaction between oxidative stress and SP-MC pathway in skin tissue. 5. Antioxidants and SP receptor blockers can partially improve the negative effect of chronic restraint stress on hair follicles. 6. Chronic restraint stress. It is an ideal model to study the relationship between psychological stress and hair follicles.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R395

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