液氮浅低温冷冻对家兔皮肤黑色素细胞影响的组织学和超微结构研究

发布时间：2018-06-29 22:55

  本文选题：液氮浅低温冷冻 + 黑色素细胞　； 参考：《滨州医学院》2010年硕士论文

【摘要】：目的: 观察液氮浅低温冷冻对家兔皮肤黑色素细胞及黑色素的组织学和超微结构的影响,以期为治疗白癜风等色素脱失性疾病提供基础理论依据。 方法: 1、实验组采用棉签蘸液氮法在家兔背部及臀部剃毛标记区反复涂擦三次(约1s),使局部结一层薄霜,待其自然融化后,再冻融2次为一周期,间隔一周再冷冻一周期,共3个周期。对照组和自身对照区同样剪毛标记,不予冷冻。 2、冷冻后每天肉眼观察皮肤组织的变化情况,包括色素的变化及毛发的生长情况等。 3、第3周和第5周取冷冻组和对照组标记区的皮肤组织,行HE染色和Masson-Fontana银染色。利用Mias病理工作站4.0拍片观察分析表皮和毛囊黑色素细胞数目及形态学变化、计数毛囊及真皮毛细血管数目;同时制备电镜切片:观察黑色素细胞及黑色素的超微结构变化。 4、统计学分析:应用SPSS13.0统计软件,分别对第3周和第5周的数据进行t检验。 结果: 1、肉眼观察:实验区自第2周起部分区域出现片状黑褐色色素沉着,并有灰黑色兔毛长出,较正常兔毛粗而黑,到第5周时色素沉着渐遍布整个冷冻标记区,同时新生毛发长至正常长度。对照组:除2处标记区出现小片状色素沉着斑及少许细软、色淡的毳毛外,余标记区皮肤均未出现色素及新生兔毛。 2、HE和Masson-Fontana银染色:实验组表皮基底层和毛囊外根鞘处黑色素细胞数目和黑色素颗粒均明显增加(P0.01),棘细胞层至颗粒层亦有黑色素颗粒沉积,真皮毛细血管数目增多(P0.01),管径扩张显著;第3周和第5周实验组各相应项目指标行成组t检验,两组四项相应指标的比较均无明显统计学差异(P0.01)。 3、电镜切片:实验组:黑色素细胞胞质内线粒体较密集、粗面内质网和游离核糖体明显增多,核切迹较少,核周有较多处于不同成熟阶段的黑素小体,以Ⅱ和Ⅲ期为主,见少量Ⅳ期的黑素体,表皮细胞内含有大量的黑色素颗粒,以基底细胞及棘细胞为显著。对照组:黑色素细胞胞核切迹较多,胞质内散在分布的内质网、核糖体、高尔基体及线粒体,胞核周围有稀疏的处于不同成熟阶段的黑色素体,以Ⅱ期为主;表皮各层中的黑色素颗粒较少,部分靠近棘层黑色素细胞的胞质内有成熟的黑色素颗粒。 结论: 1、液氮浅低温冷冻能刺激表皮和毛囊的黑色素细胞使之数目增多、功能增强及促进多巴阴性的黑色素细胞合成黑色素,因而合成大量的黑色素;同时冷冻后真皮毛细血管数目增多、管径扩张,促进了局部的血液循环和物质代谢,为新生毛发提供了营养基础,因而新生毛发黑而粗,生长旺盛。 2、本实验为治疗白癜风等色素脱失性疾病提供了理论依据,而且液氮浅低温冷冻疗法有效率高,副作用少,值得临床上广泛应用。
[Abstract]:Objective: to observe the effect of liquid nitrogen cryopreservation on the histology and ultrastructure of melanocyte and melanin in rabbit skin in order to provide basic theoretical basis for the treatment of pigment loss diseases such as vitiligo. Methods: 1. In the experimental group, three times (about 1 s) were repeatedly applied to the rabbit's back and buttocks shaving marked area by using the cotton swabs dipping liquid nitrogen method, so that a thin layer of frost was formed locally, and then frozen and thawed twice for a period after the natural thawing. Once a week and then one cycle, a total of 3 cycles. The control group and the self-control area were also marked with haircut, without freezing. 2. After freezing, the changes of skin tissue were observed with the naked eye every day after freezing. The skin tissues of the frozen group and the control group were collected at the 3rd and 5th week and stained with HE and Masson-Fontana silver staining. The number and morphology of melanocytes in epidermis and hair follicle were observed and analyzed by Mias pathological workstation 4.0. The number of hair follicles and dermal capillaries were counted. The ultrastructural changes of melanocyte and melanin were observed. 4. Statistical analysis: using SPSS 13.0 statistical software, the data of the 3rd week and the 5th week were tested by t test. Results: 1. Naked eye observation: from the second week, some areas of the experimental area appeared patch dark brown pigmentation, and the gray and black hair grew out, which was thicker and darker than the normal rabbit hair. At the 5th week, the pigmentation gradually spread over the whole freezing labeling area. At the same time, the new hair grows to normal length. In the control group, except for 2 marked areas with small pigmented spots and a few soft, pale vellus hair, The number of melanocytes and melanin granules in the basal layer of epidermis and the outer root sheath of hair follicles in the experimental group were significantly increased (P0.01), and the layer of spinous cells to granules. The layer is also deposited with melanin particles, The number of dermal capillaries increased (P0.01). There was no significant difference in the four corresponding indexes between the two groups (P0.01). 3. Electron microscope section: in experimental group: mitochondria in melanocyte cytoplasm were denser, rough endoplasmic reticulum and free ribosome were increased obviously, and nuclear notch was less. There were many melanosomes in different maturation stages around the nucleus, mainly in stage 鈪，

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