目的:研究姜黄素对博来霉素诱导肺纤维化C57BL/6小鼠Cathepsin K表达的影响。方法:博来霉素气管注射造小鼠肺纤维化模型,随机分为对照组(气管内注入0.1mL生理盐水)、模型组(气管内注入0.025U博来霉素)、姜黄素组(气管内注入0.025U博来霉素);造模后第2d,对照组和模型组给予0.5%羧甲基纤维素钠生理盐水灌胃,姜黄素组给予200 mg.kg-1.d-1姜黄素灌胃,分别于造模后第7、14、28d取材。采用H&E及Mallory染色观察肺组织病理学变化;应用免疫组学Cathepsin K蛋白的表达,经图像分析各组之间的差异。结果:姜黄素组、模型组与对照组比较都有不同程度的炎症及纤维化病灶,但是姜黄素组与对照组比较炎症及纤维化程度明显减轻;姜黄素组、模型组与对照组比较Cathepsin K表达都有不同程度增加,但是姜黄素组与对照组比较Cathep-sin K表达显著增加。结论:姜黄素可能是通过增加Cathepsin K的表达从而降解胶原而实现抗纤维化作用。

This study aimed to investigate the effect of curcumin on the expression of cathepsin K in bleomycin-induced pulmonary fibrosis C57BL/6 mice.Intratracheal application of bleomycin in C57BL/6 mice was conducted.Briefly,tracheas of anaesthetized mice were surgically exposed and 0.025 U of bleomycin diluted in sterile saline or saline only was slowly instilled.The mice were divided into three groups randomly,which were the control group,the bleomycin group,and the bleomycin plus curcumin(200 mg·kg-1 per day) group.From the second day after instillations,mice were challenged with saline or curcumin(200 mg·kg-1 per day) in 0.5% carboxymethyl cellulose via oral gavage.The mice were sacrificed at 7,14 and 28 days post-treatment after pulmonary fibrosis determination by estimating lung collagen content and lung histopathology.In addition,changes in cathepsin K protein levels in the lung were determined by immunohistochemistry,real time RT-PCR and western blotting.The results showed that cathepsin K protein levels were strongly increased in the bleomycin plus curcumin group treated mice compared to only bleomycin treated mice.Moreover,bleomycin plus curcumin group treated mice responded with a lower collagen deposition and inflammation in the lungs,which was accompanied by a significantly higher lung resistance compared to bleomycin-treated mice.In addition,mice in the bleomycin plus curcumin group were responded with a lower degree of lung fibrosis than the bleomycin-treated mice.It is concluded that curcumin via upregulation of cathepsin K expression and reduced lung collagen deposition in response to bleomycin challenge.Thereby,curcumin may partial protect against bleomycin-induced lung fibrosis.

科技部国际科技合作计划资助项目(2008DFA31970):组织蛋白酶与肺纤维化,负责人:牛建昭;教育部国家外专局高等学科创新引智计划资助项目(1307007):中药防治器官纤维化,负责人:牛建昭;北京中医药大学自主选题项目(JYBZZ-JS023):姜黄素对肺纤维化过程中组织蛋白酶的影响,负责人:黄春芳