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目的建立硬皮病小鼠模型,并了解其免疫功能特性。方法造模组用200μg/mL的BLM0.1mL注射于小鼠背部皮肤,1次/d,共3周;对照组用磷酸盐缓冲液(PBS)0.1mL同样方式处理;观察两组背部注射区皮肤的大体变化并进行组织HE染色了解组织学的变化;间接免疫荧光检测抗核抗体,免疫印迹法检测循环硬皮病相关自身抗体;对皮肤进行Masson染色,医学彩色病理图象分析系统计算免疫组织化学指数,样本碱水解法分析胶原含量。结果造模组小鼠注射部位皮肤出现皮肤增厚变硬、弹性差、毛发停止生长,皮肤和肺组织病理符合硬皮病表现,ANA阳性率50.00%(4/8),仅1例出现抗Scl-70抗体阳性。且注射区皮肤胶原染色和胶原含量均明显较对照组增多(P<0.05)。结论采用200μg/mL的BLM连续皮下注射BALB/C小鼠可建立硬皮病小鼠模型,其皮肤和肺损害具典型硬皮病组织特征,但不伴发肾脏损害,循环自身抗体的阳性率低。
Objective To establish a mouse model of scleroderma and to understand its immune function. Methods The model group was injected with 200μg / mL BLM0.1mL on the back of the mice once a day for 3 weeks. The control group was treated with 0.1mL phosphate buffered saline (PBS) in the same way. The back injection zone The general changes of the skin and tissue HE staining to understand the histological changes; indirect immunofluorescence detection of anti-nuclear antibody, Western blot detection of circulating scleroderma-related autoantibodies; the skin Masson staining, medical color pathology image analysis system to calculate the immune Histochemical index, sample alkaline hydrolysis analysis of collagen content. Results The skin of the injection site of the model group showed thickening, stiffening and poor elasticity. The hair growth stopped. The pathological appearance of the skin and lung was consistent with the scleroderma. The positive rate of ANA was 50.00% (4/8) Scl-70 antibody positive. Collagen staining and collagen content in the skin of the injected area were significantly increased compared with the control group (P <0.05). Conclusions The scleroderma mouse model can be established by continuous subcutaneous injection of 200μg / mL BLM into BALB / C mice. The skin and lung lesions are characterized by typical scleroderma but not with renal damage. The positive rates of circulating autoantibodies low.