脉冲染料激光干预小鼠黄褐斑模型的实验研究

发布时间：2018-08-27 18:28

【摘要】：目的：通过脉冲染料激光（Pulsed Dye Laser，PDL）干预治疗小鼠黄褐斑动物实验模型，检测模型皮损区皮下血管及色素变化情况，探讨PDL对黄褐斑可能的治疗意义及作用机制。方法：健康雌性昆明种小鼠40只，分为两组，其中30只为模型组，10只为正常组。均剔除其背部毛发，面积约3*2cm2。模型组予以每日肌肉注射黄体酮（0.4%），5ml/kg体质量，同时以波长为320nm的中波紫外线照射小鼠裸露皮肤，一日1次,1次60分钟，共45日，观察其脱毛部位皮肤变化情况，制作小鼠黄褐斑动物模型。建模完成后，取模型组及对照组小鼠背部皮肤，行组织学检查及皮肤丙二醛（MDA）和超氧化物歧化酶（SOD）的检测，发现模型组皮肤MDA升高和SOD降低，皮肤表皮基底层黑色素增加，验证小鼠黄褐斑模型建立成功。建模成功后，将建模后小鼠分成三组，A组：即对照组；B组：PDL治疗一次组；C组：PDL治疗二次组。以PDL干预治疗其背部皮肤，取治疗前A组、B组治疗后一周、C组治疗后一周及C组治疗后一周的对照组背部皮肤，行组织病理学检查，HE染色观察皮肤的变化，ElivisionTMplus免疫组化方法用HMB45为一抗对皮肤黑素颗粒进行标记，CD34为一抗对皮肤皮下微血管进行标记。对比皮肤颜色变化、HE染色法、免疫组化法和图像分析法观测治疗前后皮肤血管密度的变化和皮肤黑色素细胞数量及形态的改变。将资料输入计算机以SPSSl3.0软件进行统计分析，以P0.05为有统计学意义。结果： 1、30只小鼠造模后，27只存活。据所拍照片实验区域皮肤颜色逐渐加深，HMB45免疫组化染色示皮肤黑色素表达显著增多；与造模前正常组相比，皮肤组织中SOD明显降低，MDA升高。 2、以PDL对小鼠黄褐斑模型皮损区干预治疗后一周，结果显示：与对照组小鼠相比，小鼠皮下血管密度明显减少（P 0.05）；与此同时，激光干预治疗后其皮损区黑色素的表达水平也明显下降（P 0.05）；PDL两次治疗组皮下微血管及黑色素的减少更加明显。结论： 1、雌激素注射并UVB照射可建立小鼠的黄褐斑模型。 2、黄褐斑小鼠皮损区皮下血管较正常皮下血管明显增生。 3、PDL针对小鼠黄褐斑模型的皮下血管进行干预治疗，血管中的血红蛋白受热凝固，而吸收的热可能扩散至血管周围的黑色素细胞，黑色素细胞受热引起凋亡，，起到治疗黄褐斑的作用。
[Abstract]:Objective: to investigate the therapeutic significance and mechanism of PDL on chloasma in mice by using pulsed dye laser (Pulsed Dye Laser,PDL) to treat the animal model of chloasma, and to detect the changes of subcutaneous blood vessels and pigments in the lesions of the model. Methods: forty healthy female Kunming mice were divided into two groups, 30 of them were model group and 10 were normal group. The back hair was removed and the area was about 3 ~ 2 cm ~ (2). Rats in the model group were given intramuscular injection of progesterone (0.4%) with 5 ml / kg body mass, and exposed skin was irradiated with ultraviolet light (UV) at wavelength of 320nm for 60 minutes once a day for 45 days. The animal model of chloasma in mice was made. After modeling, the back skin of the model group and the control group were taken for histological examination and the detection of malondialdehyde (MDA) and superoxide dismutase (SOD) in the skin. It was found that the skin MDA increased and SOD decreased, and the melanin in the basal layer of the skin increased in the model group. The model of chloasma in mice was established successfully. After modeling successfully, the mice were divided into three groups: control group B, group 1: PDL, group C, treatment group 2. The back skin of group A was treated with PDL intervention, and the back skin of group C and group C were removed one week after treatment and one week after treatment in group C and group C respectively. Histopathological examination and HE staining were used to observe the changes of skin. The skin melanin granules were labeled with HMB45 as the first antibody and CD34 as the first antibody to mark the subcutaneous microvessels. The changes of skin vascular density and the number and morphology of melanocytes were observed before and after treatment with HE staining, immunohistochemistry and image analysis. Input the data into the computer to SPSSl3.0 software for statistical analysis, with P0.05 as statistical significance. Results: 1 27 mice survived after modeling. According to the photos taken, the skin color of the experimental area gradually deepened and the HMB45 immunohistochemical staining showed that the expression of melanin in the skin was significantly increased compared with that in the normal group before modeling. SOD in skin tissue significantly decreased the increase of MDA. 2. One week after the intervention of PDL in the lesion area of chloasma model in mice, the results showed that compared with the control group, the subcutaneous vascular density of the mice decreased significantly (P 0.05), and at the same time, the subcutaneous vascular density of the mice was significantly decreased compared with the control group (P 0.05). After laser intervention, the expression of melanin in lesions was also significantly decreased (P 0.05). The decrease of subcutaneous microvascular and melanin in PDL treatment group was more obvious than that in PDL group. Conclusion: 1. The model of chloasma in mice can be established by estrogen injection and UVB irradiation. 2. The subcutaneous vessels in the lesion area of chloasma mice are obviously proliferated compared with the normal subcutaneous vessels. The subcutaneous vessels of the plaque model were treated with intervention. The hemoglobin in the blood vessels is heated and coagulated, and the absorbed heat may spread to the melanocytes around the blood vessels. The heat of the melanocytes can induce apoptosis and play a role in the treatment of chloasma.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332;R758.42

【参考文献】