【摘要】：目的：1、建立慢性束縛應(yīng)激（Chronic restraint stress，CRS）模型，觀(guān)察心理應(yīng)激對(duì)小鼠毛囊周期的影響。2、研究P物質(zhì)（substance P, SP），氧化應(yīng)激（Oxidativestress），肥大細(xì)胞（Mast cell，MC）在心理應(yīng)激引起毛囊周期改變這一過(guò)程中的作用。3、探討脫發(fā)的發(fā)病機(jī)制，為其預(yù)防、治療及新藥開(kāi)發(fā)提供新思路和更可靠的實(shí)驗(yàn)依據(jù)。 方法：1、采用C57BL/6雄鼠建立CRS模型，與Control組比較，從肉眼和組織形態(tài)學(xué)觀(guān)察動(dòng)物毛囊周期的改變。2、在原有Control，CRS兩組基礎(chǔ)上，加入腹腔注射抗氧化劑和SP受體阻斷劑兩組，，所有小鼠隨機(jī)分為四組：Control組、CRS組、CRS+抗氧化劑組（Tempol組）、CRS+SP受體阻斷劑組（RP67580組）。使用脫毛術(shù)誘導(dǎo)小鼠背部毛囊進(jìn)入生長(zhǎng)期，選取脫毛后第9天和第19天作為時(shí)間點(diǎn)采集標(biāo)本，觀(guān)察體重、行為、皮膚顏色和毛囊組織形態(tài)的改變并對(duì)毛囊進(jìn)行評(píng)分，檢測(cè)血漿皮質(zhì)醇水平，皮膚組織中硫代巴比妥酸反應(yīng)產(chǎn)物（TBARS）水平，超氧化物歧化酶（SOD）、谷胱甘肽過(guò)氧化物酶（GSH-Px）活力；甲苯胺藍(lán)染色切片觀(guān)察皮膚肥大細(xì)胞脫顆粒比例；免疫熒光法觀(guān)察皮膚SP免疫反應(yīng)陽(yáng)性（SP+）神經(jīng)纖維數(shù)量。 結(jié)果：1、脫毛后第9天，與Control組相比，CRS組皮膚呈粉色，灰度率顯著升高（P0.05），毛囊平均分顯著降低（P0.05），生長(zhǎng)期延遲出現(xiàn)；脫毛后第19天，與Control組相比，CRS組皮膚皮膚呈黑色，灰度率和毛囊平均分均顯著降低（P0.05），退行期延遲出現(xiàn)。2、（1）與Control組相比，CRS組、Tempol組、RP67580組小鼠體重和曠場(chǎng)實(shí)驗(yàn)中3min內(nèi)運(yùn)動(dòng)距離均顯著降低（P0.05），血漿皮質(zhì)酮水平顯著升高（P0.05），提示慢性束縛應(yīng)激模型造模成功；（2）與Control組相比，CRS組皮膚組織TBARS水平顯著升高（P0.05），SOD、GSH-Px活力顯著降低（P0.05）；同CRS組相比，Tempol組和RP67580組皮膚TBARS水平顯著降低（P 0.05）、GSH-Px活力顯著升高（P0.05）；（3）脫毛后第9天，與CRS組相比，Tempol組和RP67580組皮膚呈灰色，灰度率顯著降低（P0.05），毛囊平均分顯著升高（P0.05）；脫毛后第19天，與CRS組相比，Tempol組和RP67580組皮膚呈灰色，灰度率和毛囊平均分顯著升高（P0.05），毛囊周期未見(jiàn)明顯延遲；（4）與Control組相比，CRS組皮膚SP+神經(jīng)纖維數(shù)量明顯升高（P0.05）；與CRS組相比，Tempol組和RP67580組皮膚SP+神經(jīng)纖維數(shù)量明顯降低（P0.05）。（5）同C組相比，CRS組皮膚肥大細(xì)胞脫顆粒比例明顯升高（P0.05）；與CRS組相比，Tempol組和RP67580組皮膚肥大細(xì)胞脫顆粒比例明顯降低（P0.05）。 結(jié)論：1、慢性束縛應(yīng)激可以導(dǎo)致毛囊周期的異常，表現(xiàn)為生長(zhǎng)期延遲出現(xiàn)。2、慢性束縛應(yīng)激可以引起皮膚組織氧化應(yīng)激損傷。3、慢性束縛應(yīng)激可以激活SP-MC通路，導(dǎo)致SP+神經(jīng)纖維數(shù)量上升，肥大細(xì)胞脫顆粒。4、在皮膚組織中氧化應(yīng)激與SP-MC通路之間存在交互作用。5、抗氧化劑和SP受體阻斷劑可以部分的改善慢性束縛應(yīng)激對(duì)毛囊的負(fù)性作用。6、慢性束縛應(yīng)激是研究心理應(yīng)激與毛囊之間關(guān)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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R395

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