摘要
目的探讨急性肺损伤大鼠体内基质金属蛋白酶2,9(MMP-2,MMP-9)活性的变化以及大豆胰蛋白酶抑制剂(soybean trypsin protease inhibitor,SBTI)的干预效应。方法90只SD大鼠随机分为假手术组(Sham组,n=30)、内毒素组(LPS组,n=30)、大豆胰蛋白酶抑制剂组(SBTI组,n=30)。向LPS和STBI组大鼠气管内一次性注入0.3ml含LPS(6mg.kg-1)的生理盐水复制ALI模型,假手术组大鼠则在气管内一次性注入等量生理盐水。SBTI组于造模前1天腹腔注射给予100mg.kg-1.d-1SBTI(溶于0.5ml生理盐水),假手术组和LPS组大鼠腹腔注射等量的生理盐水,3组动物同步于气管内LPS灌注后d1、d3、d7分别随机处死10只。收集肺泡灌洗液,冰浴匀浆肺组织制备匀浆液,-80℃冻存。Bradford法测定血浆蛋白含量和肺泡灌洗液中蛋白含量,计算肺通透指数;酶谱法测肺组织匀浆液和支气管肺泡灌洗液上清中MMP-2、MMP-9酶活力及免疫组织化学方法检测肺组织MMP-2、MMP-9蛋白表达情况;并进行肺组织形态学观察。结果酶谱法显示:与LPS组相比,SBTI组大鼠肺组织、支气管肺泡灌洗液中MMP-2、MMP-9活性明显下降,在d7活性变化明显,但MMP-9变化较MMP-2更明显。免疫组化显示MMP-2、MMP-9在LPS组肺组织细胞高表达。SBTI组大鼠肺组织MMP-2、MMP-9表达明显减弱。肺组织病理提示SBTI治疗组肺损伤病变局限且程度较轻,肺泡内渗出明显小于LPS组,PMN和红细胞渗出较少。结论MMP-2、MMP-9在LPS诱导大鼠急性肺损伤中发挥重要的作用,SBTI通过抑制MMPs的分泌及激活,降低基底膜的降解,减轻肺水肿及炎症反应,发挥肺保护的作用。
Aim To observe the changes of matrix metallproteinases（MMPs） activities in mice with LPS induced acute lung injury and the intervention of soybean trypsin protease inhibitor. Methods 90 male rats were randomly divided into sham group （n = 30 ） , LIPS group （n =30） and SBTI group（n =30）. LPS group and SBTI group rats were injected with a single intratracheal dose of 0. 3 ml LPS （6 mg·kg^-1 ） saline, sham group rats were injected with the same dose of sham saline. SBTI group rats were given a single dose of SBTI （ 100 mg·kg^-1·d ^-1 ） by intraperitoneal injection, the sham group and the LPS group were given the same dose of saline in the same way. 10 rats were randomly killed on day 1,3and 7 following LPS or saline instillation. The activity of MMP-2 and MMP-9 in bronchoalveolar lavage fluids （BALF） and lung homogenate was studied by zymography; the expression of MMP-2 and MMP-9 in lung tissues was studied by immunohistochemistry. LPI was measured by mean of Bradford and pathological changes of lung tissue were observed. Results Compared with LPS group, the activity of MMP-2 and MMP- 9 in BALF and lung homogenate of SBTI group decreased significantly, especially MMP-9 in day 7. The result of immunohistochemistry showed that acute lung injury caused by LPS was accompanied with increased expression of MMP-2 and MMP-9 in lung tissues. The expression of MMP-2 and MMP-9 decreased significantly in SBTI gruoup. Accordingly, lung penetrate index （LPI） was reduced and the histological changes in the lung tissues were alleviated significantly. Conclusions SBTI might take effect by means of inhibiting the expression and activity of matrix metalloproteinases in lung tissues and BALF, and sequentially, it attenuated basement membrane degradation and lung edema to protect lung from LPS induced acute lung injury.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2008年第9期1170-1173,共4页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No306727575)
长江学者和创新团队发展计划资助项目(NoIRT0413)
科技部国际合作计划资助项目(No2006DFA31230,20073813)
高等学校创新引智计划资助项目(NoB07007)
关键词
大豆胰蛋白酶抑制剂
急性肺损伤
基质金属蛋白酶
soybean trypsin protease inhibitor
acute lung injury
matrix metalloproteinases
