三转基因银屑病小鼠模型建立与表型分析

发布时间：2019-02-17 11:05

【摘要】：目的尿激酶型纤溶酶激活剂(PLAU)、尿激酶型纤溶酶激活剂受体(PLAUR)和信号传导与转录激活因子3(STAT3)是参与银屑病病理发生的重要基因。本文目的是制备皮肤特异性表达PLAU、PLAUR和STAT3三种基因的转基因小鼠,建立可进行性再现银屑病病理进程的小鼠模型。方法将PLAU、PLAUR和STAT3基因分别插入牛角蛋白5启动子(BK5)下游,构建转基因表达载体,通过显微注射法建立在皮肤组织同时高表达PLAU、PLAUR和STAT3三种基因的转基因C57BL/6J小鼠。利用PCR法鉴定转基因小鼠的基因型,Western blot检测基因表达水平,HE染色观察皮肤病理表型。结果 PLAU、PLAUR和STAT3三种基因在选定的转基因小鼠皮肤组织均有明显表达;转基因小鼠与同龄野生型小鼠相比表皮状态差,炎症明显;4月龄的转基因小鼠真皮变簿,表皮过度角化、棘层增厚,毛囊减少、发育异常、部分毛囊内未见毛干,角化不全区域内可见Munro氏小脓肿,真皮炎细胞浸润。结论建立了稳定传代的皮肤特异性表达PLAU、PLAUR和STAT3基因的转基因小鼠品系,转基因小鼠具有进行性的银屑病表型,可以作为多病因综合银屑病小鼠模型。
[Abstract]:Objective urokinase type plasminogen activator (PLAU),) urokinase type plasminogen activator receptor (PLAUR) and signal transduction and transcription activator 3 (STAT3) are important genes involved in the pathogenesis of psoriasis. The aim of this study was to prepare transgenic mice with skin specific expression of PLAU,PLAUR and STAT3 genes and to establish a mouse model that can reproduce the pathological process of psoriasis progressively. Methods PLAU,PLAUR and STAT3 genes were inserted into bovine keratin 5 promoter (BK5) downstream, and transgenic C57BL/6J mice were constructed by microinjection. PCR method was used to identify the gene expression level of genotypes of transgenic mice by, Western blot, and the skin pathological phenotypes were observed by HE staining. Results the three genes of PLAU,PLAUR and STAT3 were expressed in the skin of selected transgenic mice, and the epidermis of transgenic mice was worse than that of wild-type mice of the same age, and the inflammation of the transgenic mice was obvious compared with the wild-type mice of the same age. In 4 month old transgenic mice, the dermis became thin, the epidermis became hyperkeratosis, the spinous layer thickened, the hair follicles decreased, the development was abnormal, the hair follicles were not found in some hair follicles, the small abscess of Munro's and the infiltration of eudermatitis cells could be seen in the keratosis area. Conclusion transgenic mice with skin specific expression of PLAU,PLAUR and STAT3 genes have been established. The transgenic mice have progressive psoriasis phenotype, which can be used as a model of psoriasis mice with multiple etiological factors.
【作者单位】：北京协和医学院中国医学科学院医学实验动物研究所卫生部人类疾病比较医学重点实验室;
【基金】：国家科技支撑计划课题“基因工程大鼠模型的研发与示范”(2014BAI02B01)
【分类号】：R-332;R758.63

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