局部注射博来霉素建立硬皮病小鼠模型的实验研究

目的了解通过局部注射博来霉素(BLM)建立硬皮病小鼠模型的方法。方法选用C3H/He小鼠作为实验对象,对照组9只小鼠随机分成3组,分别在各组小鼠背部中央区皮下注射100μL0.01mol/L磷酸盐缓冲液(PBS)1次/d连续1周,2周和3周。动物模型组12只小鼠,随机分成4组,每组3只,分别背部皮下注射100μL600μg/mLBLM1次/d连续1周,2周和3周,进行皮肤组织病理学以及血清自身抗体检测验证。结果与对照组相比,连续注射3周BLM的小鼠真皮均匀硬化,皮损区表真皮厚度增加(P<0.05),羟脯氨酸含量显著增高(P<0.05),肥大细胞数目显著增加(P<0.05)伴有脱颗粒现象,TGF-β表达显著高于对照组(P<0.05)。BLM连续注射1周和2周的小鼠皮损区上述改变不明显。该小鼠模型肺泡间隔增厚,伴单一核细胞浸润,并可见肺组织小血管壁增厚。小鼠模型血清中ANA阳性,结合ENA抗体测定提示存在Scl-70抗体。硬皮病小鼠停止注射BLM后继续观察6周,皮肤硬化仍然存在。结论在C3H/He小鼠背部每日皮下注射600μg/mLBLM,连续3周可以成功建立硬皮病小鼠模型。

Establishment of Scleroderma Mouse Model Induced By Local Injection of Bleomycin

Objective To understand the mouse models of scleroderma established by local injections of Bleomycin(BLM).Methods 100 μL BLM at a concentration of 600 μg/mL was injected intracutaneusly,once per day,in the back of C3H/He mice for 1,2,3 w,respectively.The mouse models were verified by histological and pathological examination of skin specimens,lung specimens,and serum self-antibody.Control group were also set up by injection with 100 μL PBS at a concentration of 0.01 mol/L once per day for 1,2,3w,respectively.Results After 3 w local BLM injections,an intense dermal sclerosis was observed in the mouse model.Compared with that of the PBS treated mice,the dermal thickness in mice injected with BLM was increased remarkably(P<0.05) and the amount of hydroxyproline in skin was also markedly increased(P<0.05).Mast cells gradually increased in number,and showed marked degranulation(P<0.05) at the injection site.The expression level of TGF-β was much higher than that in controls(P<0.05).The specimens of the mouse lung showed thickened alveolar septa with infiltration of mononuclear cells and thickening of vascular walls.Anti-nuclear antibody were detected in serum of mouse model.Anti-Scl-70 was identified by immunoblots.The sclerotic changes of the dermis were sustained after ceasing BLM applications for at least 6 w.Conclusion The mouse models of scleroderma were successfully established in C3H/He mice by BLM administration for 3 w at concentration of 600 μg/mL.Mast cells gradually increased in number,and TGF-β is strongly expressed at the injection site of mouse models,which may be pathologically involved in fibrosis process.