ATP清除剂三磷酸腺苷双磷酸酶对实验性矽肺的干预作用  

Intervention effects of apyrase on silica-induced pulmonary fibrosis in mice

作　　者：苏程程 向国安[1] 马永强[2] 周欣 彭守春[1] 魏路清[1] 姬文婕[1] 

机构地区：[1]中国人民武装警察部队后勤学院附属平津医院呼吸与重症医学科 [2]天津市心血管重塑与靶器官损伤重点实验室,天津300162

出　　处：《中国病理生理杂志》2016年第5期792-797,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81102088;No.81441101);天津市自然科学基金资助项目(No.13JCYBJC22000;No.15JCZDJC35000);武警后勤学院附属医院重点项目(No.FYZ201510;No.FYZ201605);武警后勤学院附属医院种子基金资助项目(No.FYM201538)

摘　　要：目的:观察三磷酸腺苷双磷酸酶(apyrase,Apy)对实验性矽肺小鼠肺纤维化的干预作用及可能机制。方法:160只雄性C57BL/6小鼠,随机分为对照组、矽肺组、Apy组及溶剂组,采用经口咽吸入法给予Si O2悬浊液(50 mg/kg)建立小鼠实验性矽肺模型,对照组给予等量生理盐水。Apy组在造模的同时及造模后4 h采用经口咽吸入法给予Apy(40 mg/kg),溶剂组给予等体积的无菌生理盐水。分别于术后3 h、7 d、14 d和28 d处死小鼠,计算各组小鼠肺指数,采用生物发光法检测小鼠肺组织的ATP含量,采用HE染色和苦味酸-天狼星红染色观察肺组织的病理学变化,采用real-time PCR法检测各组肺组织中Ⅰ型胶原(ColⅠ)、Ⅲ型胶原(ColⅢ)和转化生长因子β1(TGF-β1)的mRNA表达水平,采用ELISA法测定肺泡灌洗液TGF-β1的蛋白水平。结果:与对照组相比,模型组和溶剂对照组小鼠的肺指数和胶原含量明显升高,证明矽肺模型制备成功。Apy组与溶剂组相比,肺组织的ATP含量下降,肺组织炎症表现明显减轻,炎症评分显著下降;同时肺指数、胶原容积分数及ColⅠ和ColⅢ的mRNA表达水平均显著下降。Apy可以明显下调肺组织中TGF-β1的mRNA表达水平和肺泡灌洗液中TGF-β1蛋白表达水平。结论:Apy可以明显减轻实验性矽肺小鼠肺组织的炎症反应和纤维化,该作用可能与Apy减少肺组织ATP的浓度、下调TGF-β1的表达有关。AIM： To investigate the effect of apyrase on the experimental silicosis. METHODS： C57BL/6male mice were randomly divided into control group,silica treatment group,silica ＋ apyrase group and silica ＋ NS group. A mouse model of lung fibrosis was induced by crystalline silica particles（ 50 mg / kg,via oropharyngeal instillation）,and were sacrificed at 3 h,7 d,14 d and 28 d. Apyrase was delivered by oropharyngeal aspiration at the same time and 4 h after silica challenge. The lung indexes were calculated and the concentration of ATP was detected by bioluminescent assay.The mRNA expression levels of collagen type Ⅰ（ Col Ⅰ）,collagen type Ⅲ（ Col Ⅲ） and transforming growth factor β1（ TGF-β1） were examined by real-time PCR. The protein levels of TGF-β1 in bronchoalveolar lavage fluid were measured by ELISA. RESULTS： The elevated lung index and collagen levels showed that silicosis model was established successfully. Compared with silica group,apyrase treatment significantly alleviated silica-induced inflammation,reduced inflammation score on day 7,and decreased the lung index,collagen volume fraction and the mRNA expression of Col Ⅰand Col Ⅲon day 28. Treatment with apyrase effectively down-regulated the mRNA levels of TGF-β1 in the lung tissues and TGF-β1protein levels in bronchoalveolar lavage fluid on day 7. CONCLUSION： Apyrase attenuates the pulmonary inflammation and fibrosis of silicosis,which may be related with down-regulation of ATP and TGF-β1 in the lung tissues.

关 键 词：矽肺 三磷酸腺苷双磷酸酶 转化生长因子Β1 

分 类 号：R135.2[医药卫生—劳动卫生] R363[医药卫生—公共卫生与预防医学]