乌司他丁对矽肺大鼠肺组织病理状态与水通道蛋白1及基质金属蛋白酶表达的影响

摘要

目的:研究乌司他丁对矽肺大鼠肺组织病理状态与水通道蛋白1(AQP-1)及基质金属蛋白酶2(MMP-2)、MMP-9表达的影响.方法:取Wistar大鼠分为正常对照组、模型组和乌司他丁高、中、低剂量组(50×104、10×104、2×104 u/kg),每组20只.除正常对照组外,其余各组大鼠采用非暴露式气管内注入法灌注40 mg/mL的SiO2混悬液1 mL建立矽肺模型;建模给药前1 h ip相应剂量的乌司他丁,连续给药3 d.采用苏木精-伊红染色观察各组大鼠肺组织矽结节病理情况;免疫荧光法检测肺组织中AQP-1的表达;实时荧光定量聚合酶链式反应法检测肺组织中MMP-2、MMP-9 mRNA的表达;Western blot法检测肺组织中MMP-2、MMP-9蛋白的表达.结果:模型组大鼠肺组织可见明显矽结节出现,肺组织结构破坏严重;乌司他丁各剂量组大鼠可见含尘巨噬细胞灶,但肺纤维化程度较模型组明显减轻.与正常对照组比较,其余各组大鼠肺组织中AQP-1表达均降低,MMP-2、MMP-9 mRNA及蛋白表达均增强(P<0.05);与模型组比较,乌司他丁各剂量组大鼠肺组织中AQP-1表达均增加,MMP-2、MMP-9 mRNA及蛋白表达均减弱(P<0.05),其中高、中剂量组改善效果较低剂量组明显(P<0.05).结论:乌司他丁能缓解矽肺模型大鼠病理状态,可上调肺组织中AQP-1表达和下调MMP-2、MMP-9表达.%OBJECTIVE:To study the effects of ulinastatin on pathological state of lung tissue and aquaporin 1(AQP-1)and matrix metalloproteinases (MMP-2),MMP-9 expressions in silicosis rats. METHODS:Wistar rats were divided into normal con-trol group,model group,ulinastatin high-dose,medium-dose,low-dose groups(50×104,10×104,2×104 u/kg),20 in each group. Except for normal control group,rats in other groups were given 40 mg/mL SiO2 suspension 1 mL by non-exposed endotracheal per-fusion to induce silicosis model;1 h before modeling administration,corresponding doses of ulinastatin were intraperitoneally in-jected,for 3 d. Hematoxylin-eosin staining was used to observe silicon nodules in lung tissue of rats in each group;immunofluores-cence method was used to detect AQP-1 expression in lung tissue;real-time fluorescence quantitative polymerase chain reaction method was used to detect MMP-2,MMP-9 mRNA expressions in lung tissue;Western blot method was used to detect MMP-2, MMP-9 protein expressions in lung tissue. RESULTS:There were obvious silicon nodules and serious structural damage in lung tis-sue in model group,ulinastatin groups showed macrophage foci visible dust,but pulmonary fibrosis was obviously reduced. Com-pared with normal control group,AQP-1 expression in lung tissue in other groups were reduced,MMP-2,MMP-9 mRNA and pro-tein expressions were enhanced (P<0.05). Compared with model group,AQP-1 expression in lung tissue in ulinastatin groups were increased,MMP-2,MMP-9 mRNA and protein expressions were decreased (P<0.05),in which improvement effects in high-dose,medium-dose groups were superior to low-dose group(P<0.05). CONCLUSIONS:Ulinastatin can reduce the patholog-ical state of silicosis rats,increase AQP-1 expression and decrease MMP-2,MMP-9 expressions.