吸入染毒纳米级炭黑气溶胶致小鼠肺炎性损伤的研究  

Study on the pulmonary inflammatory effects induced by inhalation exposure to nanoscale carbon black aerosol in mice

作　　者：李园园[1] 高峰[1] 解秋艳[2] 牛勇[2] 孟涛[2] 张荣[3] 陈雯[1] 郑玉新[2] 

机构地区：[1]中山大学公共卫生学院预防医学系, 广州510080 [2]中国疾病预防控制中心职业卫生与中毒控制所 中国疾病预防控制中心化学与健康安全重点实验室 [3]中国疾病预防控制中心职业卫生与中毒控制所 中国疾病预防控制中心化学与健康安全重点实验室 河北医科大学公共卫生学院毒理教研室

出　　处：《中华预防医学杂志》2014年第12期1098-1103,共6页Chinese Journal of Preventive Medicine

基　　金：国家自然科学基金（81130050）;中国博士后基金（2013M540124）

摘　　要：目的：通过动式吸入纳米级炭黑气溶胶建立小鼠经呼吸道染毒模型，探讨吸入炭黑气溶胶引起小鼠肺组织结构改变和早期炎性反应。方法使用动式吸入染毒装置发生炭黑气溶胶，将其导入染毒室进行吸入毒性染毒。使用扫描电子显微镜和透射电子显微镜表征炭黑颗粒的粒径。将60只9周龄雄性BALB／c小鼠按体重用随机数字表法分为4组，即7 d染毒组和对照组、14 d染毒组和对照组，每组各15只。染毒组经动式吸入染毒柜使小鼠暴露在浓度约为（29．33±9．10） mg／m3炭黑气溶胶中6h／d，分别连续染毒7d和14d。7d染毒组和14d染毒组小鼠在末次染毒后24h处死，同时处死对照组小鼠。解剖收集小鼠气管、肺脏、肝脏、肾脏、脾脏。肺组织采用苏木素－伊红（ HE）染色，光镜下观察肺组织的病理学改变；通过免疫组织化学方法分析肺组织中细胞因子白细胞介素－8（ IL－8）的表达水平；运用透射电子显微镜观察肺组织的超微结构。结果14 d染毒组和对照组肺脏器系数分别为0．79±0．06、0．61±0．03（t＝6．26，P＜0．01），脾脏器系数分别是0．51±0．06、0．39±0．04（t＝4．23，P＜0．01），其余各组各脏器系数的差异无统计学意义（P＞0．05）。组织病理学显示染毒7 d、14 d小鼠肺支气管壁和肺泡内均有炭黑颗粒物沉积，染毒14 d小鼠肺组织内黑色炭黑颗粒沉积明显增多，肺组织结构表现为纤毛破坏、肺泡壁破坏严重、不完整、炎细胞浸润多、充血等。免疫组织化学检查发现肺组织中炎性因子 IL－8强度7 d 染毒组（0．272±0．011）、14 d 染毒组（0．422±0．065）高于14 d对照组（0．188±0．041）（F＝31．89，P＜0．01）。电镜结果显示对照组小鼠肺组织视野较清晰，细胞器完整；染毒组肺组织巨噬细胞溶酶体中出现吞噬颗粒，其电子密度与炭黑颗粒一致。结论吸入染Objective To explore the carbon black induced effects of lung morphology and pro-inflammation in mice, based on the carbon black aerosol dynamic inhalation exposure model.Methods The carbon black aerosol generated by dynamic inhalation device was imported exposure chamber to mice. Scanning electron microscopy （ SEM） and transmission electron microscopy （ TEM） were used to observe the characters of carbon black.Sixty 9-week-old male BALB/c mice were randomly divided into two control groups, 7 d exposure group and 14 d exposure group.The numbers of four groups of animals were 15, respectively.Mice were exposed to carbon black in the inhalation chamber at （ 29.33 ±9.10 ） mg/m3 for 6 h/d for continuous exposure 7 d and 14 d, respectively.After 7 d and 14 d exposure, the mice were sacrificed after the last exposure for 24 h.Control mice were killed at 7 d and 14 d.The trachea, lungs, liver, kidneys, and spleen tissues were separated and weighted.Hematoxylin and eosin （HE） staining was used to observe pathological changes of lung by light microscopy.Pulmonary interleukin-8 （ IL-8） expression 〈br〉 was analyzed by immunohistochemistry.Transmission electron microscopy was used to observe the ultra structure of lung tissue.Results After 14 d exposure carbon black, the lung coefficient was increased in exposure group compared with control （0.61 ±0.03 vs 0.79 ±0.06, t =6.26, P 〈0.01）.The spleen coefficient were higher than control （ 0.39 ±0.04 vs 0.51 ±0.06, t =4.23, P 〈0.01 ） .Other organ coefficients were no significant difference between CB group and control group.Histopathology displayed carbon black particles were deposited in the alveoli and lung bronchial wall in 7 d and 14 d groups.The black carbon particles were deposited within the lung tissue of mice in 14 d group.There were cilia damage, serious damage to the alveolar wall, inflammatory cell infiltration and more hyperemia in 14 d group. Immunohistochemistry showed the level of IL-8 in 7 d（0.272 ±0.011） and 14 d （0.

关 键 词：炭黑 纳米粒 动式吸入染毒 病理学 

分 类 号：R114[医药卫生—卫生毒理学]