摘要
目的 探讨乌司他丁对光气所致大鼠急性肺损伤的保护作用及与基质金属蛋白酶-9（MMP-9）的关系.方法 选取健康成年SD雄性大鼠64只,随机分为染毒组[光气染毒组（B1组）、生理盐水染毒组（B2组）、地塞米松治疗组（B3组）、乌司他丁治疗组（B4组）]和非染毒组[空气对照组（A1组）、生理盐水对照组（A2组）、地塞米松对照组（A3组）、乌司他丁对照组（A4组）],每组8只.各组大鼠均在注射相应的药物（地塞米松、生理盐水或乌司他丁）1 h后,非染毒组吸入新鲜空气5 min,染毒组吸人纯度为100%的 8.33 mg/L光气5 min.6 h后取各组大鼠的肺组织行HE染色,计算肺湿/干重量比值（W/D）,检测肺泡灌洗液（BALF）中总蛋白含量及白细胞计数,利用免疫组织化学法检测肺组织中MMP-9蛋白表达,酶联免疫吸附法检测血清中MMP-9活力以及明胶酶谱法检测BALF中MMP-9酶原含量.结果 与A1、A2、A3、A4组比较,B1和B2组大鼠肺W/D、BALF中蛋白含量和白细胞计数均明显升高,差异有统计学意义（P〈0.01）.与B1和B2组比较,B3和B4组大鼠肺W/D、BALF中蛋白含量及白细胞计数明显下降,差异有统计学意义（P〈0.01）.B1和B2组大鼠肺泡壁明显充血、增厚,肺泡壁和肺间质内可见较多白细胞浸润以及肺泡结构的破坏.B3和B4组大鼠肺泡结构较为清晰,肺泡壁稍增厚,伴少量炎性细胞浸润.非染毒组大鼠肺、支气管组织中MMP-9蛋白表达均呈弱阳性,B1组和B2组MMP-9蛋白表达呈强阳性,B3组和B4组肺组织中MMP-9蛋白表达明显减弱,恢复至正常肺组织的弱阳性表达水平.与A1、A2、A3、A4组比较,B1和B2组大鼠血清中MMP-9活力升高,差异有统计学意义（P〈0.01）,与B1和B2组比较,B3和B4组血清中MMP-9活力均明显下降,差异有统计学意义（P〈0.01）.B1、B2组BALF中MMP-9酶原含量明显高于B3、B4组,差异有统计学意义（P〈0.05）.结论 乌司他丁可以抑制MMP-9蛋白表达的上调,对于光气所致的ALI具有保护作用.
Objective To observe the protective mechanism of ulinastatin on mice with acute lung injury induced by exposure to phosgene and its relationship to the expressions of matrix metauoproteinase-9 （MMP-9） in the lung tissues. Methods Sixty-four healthy male SD rats were randomly divided into two groups： the experimental group and the control group.32 rats in the experiment group were randomly subdivided into four groups： rats with phosgene exposure group, rats with phosgene exposure after saline injected group, rats with phosgene exposure after dexamethasone injected group. 32 rats in the control group were randomly subdivided into four groups： rats with air exposure group, pretreated with ulinastatin before air exposure group, pre-treated with saline before air exposure group, pretreated with dexamethasone before air exposure group, 8 animals in each group. After pretreated with the same dose of ulinastatin, saline, dexamethasone respectively, 32 rats in the control groups were exposed to the air on the same condition respectively for 5 min. While after pretreated with the same dose of ulinastatin, saline, dexamethasone respectively, 32 rats in the experiment groups were exposed to the phosgene which the concentration was 8.33 mg/L and with 100% purity for 5 min. The lung wet/dry （W/D） weight ratio was calculated, and total protein content and BALF leukocyte count were detected. The immunohistochemistry was used to detect lung tissue protein expression MMP-9 while enzyme-linked immunosorbent method was employed to detect MMP-9 in serum levels and enzyme original gelatinases spectrum method to detect BALF MMP-9 enzyme original content. Results Compared with A1, A2, A3, A4 group, the lung W/D, BALF of protein content and WBC count in B1 and B2 group rats were significantly increased, and the difference was statistically significant （P〈0.01）. There was statistically significant difference in lung W/D, BALF of protein content and white blood cell count between B1,B2 group and the B3 and B4 rats （P〈0.01）. Histological experimental results showed marked hyperemia of alveolar walls, thickening in the lungs, alveolar walls and stroma cells infiltrating and more visible alveolar structure damage in B1 and B2 rats while the alveolar structure, the alveolar walls were clear and slightly thickened with inflammatory cells in B3 and B4 rats. Immunohistochemical result showed that the individual rats, lung and bronchus organization MMP-9 protein were weakly positive, B1 and B2 group MMP-9 protein expression was strongly positive,B3 group and the group MMP B4 lung tissue protein expression-9 weakens, restored to the normal lung tissue of weakly positive expression level. ELISA and gelatinases spectrum testing showed B1 and B2 rats, serum MMP-9 enzyme activity and content increased compared with A1, A2, A3, A4 group, the differences were statistically significant （P〈 0.01）, and B1, B2 group compared with the B3 and serum B4 group MMP-9 enzyme activity and the differences were obviously decreased, with statistically significant difference（P〈0.01）. Conclusions Ulinastatin has protective effect on phosgene-induced ALI. Ulinastatin can inhibit the up-regulation of expression of MMP-9.
出处
《中华劳动卫生职业病杂志》
CAS
CSCD
北大核心
2010年第7期498-504,共7页
Chinese Journal of Industrial Hygiene and Occupational Diseases
基金
江苏省教育厅开放课题资助项目（KJS050004）
上海市科委科研项目支助（54119610）
上海市公共卫生重点学科建设项目（08gwzx0404）
关键词
光气
急性肺损伤
基质金属蛋白酶
Phosgene
Acute lung injury
Matrix metalloproteinases
作者简介
通讯作者：申捷，E-mail：j1999sh@yahoo．com．cn
