染矽尘大鼠克拉拉细胞蛋白和表面活性蛋白D表达变化  

Dynamic changes in expression of clara cell protein and surfactant protein-D expressions in lung tissues and bronchoalveolar lavage fluid of silica-treated rats

作　　者：张海鹏[1] 王瑞[1] 王辉[1] 张玮[1] 

机构地区：[1]山东省职业卫生与职业病防治研究院,济南250062

出　　处：《中华劳动卫生职业病杂志》2014年第3期168-172,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金（30872094）;山东省科技攻关项目（2013GSF11828）

摘　　要：【摘要】目的探讨染矽尘大鼠早期肺组织以及支气管肺泡灌洗液（BALF）中克拉拉细胞蛋白（CC16）和表面活性蛋白D（SP-D）的表达变化。方法将84只Wistar大鼠随机分成对照组与染尘组，每组42只。染尘组又分为染尘3、7、14、21、28和60d组。采用一次性气管内注入矽尘悬浮液法（100mg／m1）对大鼠进行染尘，染尘后于3、7、14、21、28和60d分别处死大鼠，取其肺组织并收集BALF。采用免疫组化法检测不同时间点肺组织中SP-D和CC16的表达；采用酶联免疫吸附法（ELISA）检测BALF中SP-D和CC16的浓度。结果免疫组化法结果显示，CC16和SP．D均在肺部细胞内阳性表达。ELISA结果显示，染尘7、14、21、28、60d组大鼠BALF中CC16含量分别为（8．14±0．70）（7．15±0．66）（7．00±0．69）（6．34±0．59）（5．27±0．49）ng／L，随着染矽尘时间的不断延长，CC16表达量也随着减少，呈负相关关系（ra=-0．953，P〈0．01）。与对照组比较，各染尘组CC16表达明显减少，差异有统计学意义（P〈0．05）。染尘7d组大鼠BALF中SP—D的含量表达明显升高，染尘14d组SP-D含量达到最高峰，之后其含量逐渐降低。染尘7、14、21d组大鼠BALF中SP-D含量分别为（12．20±1．57）（14．41±0．65）（12．18±0．74ng／L），明显高于对照组，差异均有统计学意义（P〈0．05）。结论矽尘能够导致大鼠肺组织和BALF中CC16和SP-D蛋白表达发生改变，随染尘时间的不断延长，CC16的含量逐渐减少，SP-D的含量先增加后降低。Objective To investigate the dynamic changes in the expression of clara cell protein （CC16） and surfactant protein D （SP-D） in the lung tissues and bronchoalveolar lavage fluid （BALF） of silicatreated rats. Methods Eighty-four Wistar rats were randomly divided into control group （n=42） and silica group （n=42）. The silica group was subsequently divided into 3, 7, 14, 21, 28, and 60 d subgroups. The silicotic model was made by instilling silica suspension directly through the trachea into rat lungs.At 3, 7, 14, 21, 28, and 60 d after silica instillation, 8 rats in each group were sacrificed and their lung tissues and BALF were collected. The expression of SP-D and CCI6 in lung tissues was detected by immunohistochemistry. Enzyme-linked immunosorbent assay （ELISA） was used to measure the levels of SP-D and CC16 in BALF. Results The immunohistochemical assay indicated that CC16 and SP-D were expressed in lung cells. The ELISA found that in 7, 14, 21, 28, and 60 d silica subgroups, the content of CC16 in rat BALF was 8.14±0.70, 7.15±0.66, 7.00± 0.69, 6.34 ±0.59, and 5.27 ±0.49 ng/L, respectively; CC16 expression decreased gradually with the silica exposure time prolonged, indicating a negative correlation （ra=-0.953, P〈0.01 ）. Compared with the control group, all silica subgroups had significantly decreased CC16 levels （P〈0.05）. The content of SP-D in BALF was 12.20 ±1.57, 14.41 ±0.65, and 12.18 ±0.74 ng/L, respectively, in the 7, 14, and 21 d silica subgroups, significantly higher than that in the control group （P〈0.05）. Conclusion The dynamic changes in SP-D and CC16 protein levels in the lung tissues and BALF of rats could be induced by silica exposure and are related to silica exposure time. With the extension of silica exposure, CC16 levels are gradually reduced, while the SP-D levels first increase and then fall.

关 键 词：二氧化硅 支气管肺泡灌洗液 克拉拉细胞蛋白 表面活性蛋白D 

分 类 号：R135.2[医药卫生—劳动卫生]