建立硬皮病小鼠模型的实验研究

Experimental study of establishing mouse model for scleroderma

目的 建立硬皮病小鼠模型。方法 取雌性BALB/C和C3H小鼠各 14只 ,平均分为造模组1和对照组。造模组1用 2 0 0 μg/ml的博来霉素 (bleomycin ,BLM ) 0 1ml注射于小鼠背部皮肤 ,每日 1次 ,共 3周 ;对照组用磷酸盐缓冲液 (PBS) 0 1ml作背部皮内注射 ,每日 1次 ,共 3周 ;观察两组皮肤和肺组织学改变 ,应用医学彩色病理图象分析系统测定真皮厚度 ;用光电比色法测定皮肤中羟脯氨酸和蛋白的含量来推断胶原含量 ;进行Ⅰ、Ⅲ型胶原和转化生长因子 (TGF) β1免疫组织化学染色 ,计算了免疫组织化学指数。用SPSS软件对两组的检测值作统计学分析。结果 造模组1小鼠的皮肤厚度较对照组明显增厚 (P <0 0 0 1)。皮肤胶原含量比对照组显著增高 (P <0 0 5 )。皮肤Ⅰ、Ⅲ型胶原免疫组织化学指数比对照组明显增加 (P <0 0 0 1)。皮肤TGF β1免疫组织化学指数比对照组亦有极明显增加 (P <0 0 0 1)。结论 2 0 0

Objective To establish a mouse model for scleroderma.Methods Forteen BALB C and 14 C3H female mice were averagely divided into model 1 and controls.Daily 0 1 ml BLM at a concentration of 200 μg/ml was injected intracutaneusly into the backs of model 1 mice for 3 weeks,and 0 1 ml solution of PBS were injected intracutaneusly into the backs of control mice for 3 weeks.Observing the histological change of skin and lungs was made and measuring the thickness of dermis was performed with a medical analysis system of the color picture,determined the collagenic quantity was done with photoelectric colorimetry,and calculating the immunohistochemical index of collagen type Ⅰ and Ⅲ and transforming growth factor β 1 (TGF β 1) in the skin lesions from the mouse model and control was done.SPSS was used to finish the statistical analysis of the detective value from model 1 and controls.Results In the skin of model mice,the thickness of dermis markedly thickened ( P <0 001),collagenic quantity obviously increased ( P <0 05),and immunohistochemical index of collagen type Ⅰ and Ⅲ and TGF β 1 all markedly increased ( P <0 001) more than those of controls.Conclusion The animal model of scleroderma can be successfully established in BALB C and C3H mice with BLM at a concentration of 200 μg/ml.