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

Effects of Ulinastatin on Pathological State of Lung Tissue and Aquaporin 1 and Matrix Metalloproteinase Expressions in Silicosis Rats

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(50×10~4、10×10~4、2×10~4u/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SiO_2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 m RNA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 m RNA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 m RNA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 control group,model group,ulinastatin high-dose,medium-dose,low-dose groups（50×10^4,10×10^4,2×10^4u/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 perfusion to induce silicosis model;1 h before modeling administration,corresponding doses of ulinastatin were intraperitoneally injected,for 3 d. Hematoxylin-eosin staining was used to observe silicon nodules in lung tissue of rats in each group;immunofluorescence 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 tissue in model group,ulinastatin groups showed macrophage foci visible dust,but pulmonary fibrosis was obviously reduced. Compared with normal control group,AQP-1 expression in lung tissue in other groups were reduced,MMP-2,MMP-9 mRNA and protein 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 pathological state of silicosis rats,increase AQP-1 expression and decrease MMP-2,MMP-9 expressions.