【摘要】：目的： 通過(guò)脈沖染料激光（Pulsed Dye Laser，PDL）干預(yù)治療小鼠黃褐斑動(dòng)物實(shí)驗(yàn)?zāi)Ｐ停瑱z測(cè)模型皮損區(qū)皮下血管及色素變化情況，探討PDL對(duì)黃褐斑可能的治療意義及作用機(jī)制。 方法： 健康雌性昆明種小鼠40只，分為兩組，其中30只為模型組，10只為正常組。均剔除其背部毛發(fā)，面積約3*2cm2。模型組予以每日肌肉注射黃體酮（0.4%），5ml/kg體質(zhì)量，同時(shí)以波長(zhǎng)為320nm的中波紫外線照射小鼠裸露皮膚，一日1次,1次60分鐘，共45日，觀察其脫毛部位皮膚變化情況，制作小鼠黃褐斑動(dòng)物模型。建模完成后，取模型組及對(duì)照組小鼠背部皮膚，行組織學(xué)檢查及皮膚丙二醛（MDA）和超氧化物歧化酶（SOD）的檢測(cè)，發(fā)現(xiàn)模型組皮膚MDA升高和SOD降低，皮膚表皮基底層黑色素增加，驗(yàn)證小鼠黃褐斑模型建立成功。建模成功后，將建模后小鼠分成三組，A組：即對(duì)照組；B組：PDL治療一次組；C組：PDL治療二次組。以PDL干預(yù)治療其背部皮膚，取治療前A組、B組治療后一周、C組治療后一周及C組治療后一周的對(duì)照組背部皮膚，行組織病理學(xué)檢查，HE染色觀察皮膚的變化，ElivisionTMplus免疫組化方法用HMB45為一抗對(duì)皮膚黑素顆粒進(jìn)行標(biāo)記，CD34為一抗對(duì)皮膚皮下微血管進(jìn)行標(biāo)記。對(duì)比皮膚顏色變化、HE染色法、免疫組化法和圖像分析法觀測(cè)治療前后皮膚血管密度的變化和皮膚黑色素細(xì)胞數(shù)量及形態(tài)的改變。將資料輸入計(jì)算機(jī)以SPSSl3.0軟件進(jìn)行統(tǒng)計(jì)分析，以P0.05為有統(tǒng)計(jì)學(xué)意義。 結(jié)果： 1、30只小鼠造模后，27只存活。據(jù)所拍照片實(shí)驗(yàn)區(qū)域皮膚顏色逐漸加深，HMB45免疫組化染色示皮膚黑色素表達(dá)顯著增多；與造模前正常組相比，皮膚組織中SOD明顯降低，MDA升高。 2、以PDL對(duì)小鼠黃褐斑模型皮損區(qū)干預(yù)治療后一周，結(jié)果顯示：與對(duì)照組小鼠相比，小鼠皮下血管密度明顯減少（P 0.05）；與此同時(shí)，激光干預(yù)治療后其皮損區(qū)黑色素的表達(dá)水平也明顯下降（P 0.05）；PDL兩次治療組皮下微血管及黑色素的減少更加明顯。 結(jié)論： 1、雌激素注射并UVB照射可建立小鼠的黃褐斑模型。 2、黃褐斑小鼠皮損區(qū)皮下血管較正常皮下血管明顯增生。 3、PDL針對(duì)小鼠黃褐斑模型的皮下血管進(jìn)行干預(yù)治療，血管中的血紅蛋白受熱凝固，而吸收的熱可能擴(kuò)散至血管周圍的黑色素細(xì)胞，黑色素細(xì)胞受熱引起凋亡，，起到治療黃褐斑的作用。
[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.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R-332;R758.42

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