对硬皮病小鼠模型影响的实验研究

发布时间：2018-05-05 06:29

本文选题：硬皮病 + UVA_1　；参考：《第三军医大学》2008年硕士论文

【摘要】： 一、研究背景和目的硬皮病是一种病因不明、发病机制尚不明确、主要以皮肤或内脏器官纤维化、血管异常病变和免疫异常激活为主要特点的自身免疫性结缔组织病。其中纤维化是硬皮病发病过程中最突出的特点,也是严重影响病人生活质量、甚至危及病人生命的主要威胁。研究表明:UVA能够穿透表皮、真皮,甚至深达皮下,对局部照射部位的组织、细胞、血管以及DNA等产生影响,从而起到调节机体状态和治疗疾病的作用。已经证实UVA具有抗炎、免疫调节及改善局部微循环等作用,在皮肤科其他疾病的治疗上已经应用多年。近几年,多个研究已经表明应用UVA1治疗硬皮病患者取得了肯定疗效,但对UVA1治疗硬皮病的作用机制研究关注较少。本研究首次在建立博莱霉素硬皮病小鼠模型的基础上,观察UVA1照射前后硬皮病小鼠局部注射部位皮肤的外观变化、病理变化、羟脯氨酸含量及胶原含量变化,并测定TGFβ1、TGFβ2和decorin等与硬皮病发病机制密切相关的细胞因子的变化,试图探索UVA1治疗硬皮病的可能机制,为进一步理解硬皮病的发病机制和探索新的治疗手段提供重要的参考依据。二、方法用300μg/ml的博莱霉素每日注射BALB/c小鼠背部皮肤,连续28天,进行模型构建;之后,通过观察注射部位皮肤外观变化、皮损及肺部组织的组织病理变化、羟脯氨酸含量及胶原含量变化来验证模型构建是否成功。用波长365nm UVA1照射博来霉素诱导的硬皮病小鼠模型,观察照射前后局部皮肤变化,组织病理变化、羟脯氨酸含量及胶原含量变化,并用免疫组化法及RT-PCR测定decorin、TGFβ1及TGFβ2等因子在各组小鼠皮肤的变化。三、结果 1.从皮肤外观变化、皮损及肺部组织的组织病理表现、羟脯氨酸含量及胶原含量验证了硬皮病小鼠模型构建是成功的。 2. UVA1照射前后,照射组小鼠与模型组小鼠相比,局部硬化皮肤明显软化,与皮下粘连明显减轻;羟脯氨酸含量及胶原含量显著降低(P0.05);病理示:真皮层变薄,胶原纤维排列疏松,数量减少;从局部皮肤的外观变化、皮肤组织病理和羟脯氨酸含量与胶原含量的变化三个方面均显示出:UVA1照射可以有效改善硬皮病小鼠的硬化皮肤。 3.通过测定UVA1照射前后TGFβ1、TGFβ2和decorin的变化,显示出:在UVA1照射前的硬皮病小鼠皮损内,TGFβ1、TGFβ2呈阳性高表达,decorin则呈现低表达;在UVA1照射后的硬皮病小鼠皮损内,TGFβ1、TGFβ2呈较显著的下调表达,decorin则呈上调表达。四、结论 1.不仅从外观上,而且从组织病理、羟脯氨酸含量和胶原含量上均显示出:UVA1光疗可显著改善硬皮病小鼠的硬化皮肤,使其明显软化,弹性恢复,并使其羟脯氨酸含量和胶原含量有意义的下降,胶原纤维数目减少,排列疏松。 2.通过观察UVA1照射前后TGFβ1、TGFβ2和decorin的变化,推测UVA1对硬皮病小鼠模型影响的机制可能与UVA1照射后下调TGFβ1、TGFβ2和上调decorin有关。
[Abstract]:First, research background and purpose
Scleroderma is a kind of autoimmune connective tissue disease which is mainly characterized by fibrosis of the skin or viscera, abnormal vascular lesions and activation of immune abnormality, among which fibrosis is the most prominent characteristic of scleroderma, and it is also a serious influence on the quality of life of the patients and even endanger the disease. The main threat of human life. Studies have shown that UVA can penetrate the epidermis, dermis, even deep to the subcutaneous, and affect the tissues, cells, blood vessels and DNA of the local irradiated parts, which can regulate the state of the body and treat the disease. It has been proved that UVA has the effect of anti-inflammatory, immunoregulation and improvement of local microcirculation, in the Department of dermatology. The treatment of his disease has been used for many years. In recent years, many studies have shown that the application of UVA1 in the treatment of scleroderma has achieved positive results, but there is less attention in the study of the mechanism of UVA1 for scleroderma.
On the basis of the establishment of a mouse model of bleomycin scleroderma for the first time, the changes in the skin appearance, pathological changes, the changes of hydroxyproline content and the content of collagen in the local injection site of scleroderma before and after UVA1 irradiation were observed, and the changes of cytokines, such as TGF beta 1, TGF beta 2 and decorin, were measured, and the changes in the cell factors closely related to the pathogenesis of scleroderma were measured. The purpose of this study is to explore the possible mechanism of UVA1 in the treatment of scleroderma so as to provide an important reference for further understanding the pathogenesis of scleroderma and exploring new therapeutic methods.
Two, method
300 g/ml of bleomycin was injected into the back skin of BALB/c mice daily for 28 days and the model was constructed. After that, the skin appearance changes, skin lesions and pathological changes of the lung tissue were observed, the content of hydroxyproline and the content of collagen were observed to verify the success of the model construction. The wavelength of 365nm UVA1 was used to irradiate bleomycin. In the induced scleroderma mouse model, the changes of local skin and histopathology, the content of hydroxyproline and the content of collagen were observed before and after irradiation, and the changes of decorin, TGF beta 1 and TGF beta 2 were measured by immunohistochemistry and RT-PCR.
Three, the result
1. from the changes of skin appearance, histopathological findings of skin and lung tissue, hydroxyproline content and collagen content, it is proved that the construction of scleroderma mouse model is successful.
2. UVA1 irradiation, compared with the model group, the mice in the irradiated group were obviously softened and decreased with the subcutaneous adhesion, the content of hydroxyproline and the content of collagen decreased significantly (P0.05), and the pathology showed that the dermis became thinner, the collagen fibers were loosely arranged and the quantity decreased; the appearance of the skin, the histopathology of the skin and the hydroxyproline from the local skin. Three aspects of acid content and collagen content change showed that UVA1 irradiation could effectively improve scleroderma in scleroderma mice.
3. by measuring the changes of TGF beta 1, TGF beta 2 and decorin before and after UVA1 irradiation, it showed that in the skin lesions of scleroderma mice before UVA1 irradiation, TGF beta 1, TGF beta 2 showed positive high expression, decorin showed low expression, and TGF beta 1 and TGF beta 2 were significantly downregulated in the skin lesions of scleroderma after UVA1 irradiation, and the decorin was up expression.
Four. Conclusion
1. not only the appearance, but also the histopathology, the content of hydroxyproline and the content of collagen showed that UVA1 phototherapy could significantly improve the hardened skin of scleroderma mice, make it soften obviously, restore the elasticity, decrease the content of hydroxyproline and collagen, decrease the number of collagen fibers, and arrange the arrangement loosely.
2. by observing the changes of TGF beta 1, TGF beta 2 and decorin before and after UVA1 irradiation, it is presumed that the mechanism of UVA1 on scleroderma mouse model may be related to the downregulation of TGF beta 1, TGF beta 2 and up regulation of decorin after UVA1 irradiation.

【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R593.25;R-332

【引证文献】