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[小鼠吸入纳米级炭黑气溶胶所致肺部炎症效应的研究]

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

作者信息

Li Yuanyuan, Gao Feng, Xie Qiuyan, Niu Yong, Meng Tao, Zhang Rong, Chen Wen, Zheng Yuxin

机构信息

School of Public Health, Sun Yat-sen University, Guangzhou 510080, China.

Email:

出版信息

Zhonghua Yu Fang Yi Xue Za Zhi. 2014 Dec;48(12):1098-103.

Abstract

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/m(3) 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 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 (0.422 ± 0.065) exposure group were higher than control group in 14 d (0.188 ± 0.041) , F = 31.89, P < 0.01. TEM showed that the lung tissue vision was clear and organelle integrity in the control group. The particles appeared in lung tissue macrophage lysosomes in exposure group, the electron density was consistent with the carbon black particles.

CONCLUSION

The dynamic carbon black particles exposure can affect the lung and spleen coefficient, damage integrity of lung morphology and induce inflammation in mice.

摘要

目的

基于炭黑气溶胶动态吸入暴露模型,探讨炭黑对小鼠肺形态及促炎作用的影响。

方法

通过动态吸入装置产生的炭黑气溶胶导入暴露箱作用于小鼠。采用扫描电子显微镜(SEM)和透射电子显微镜(TEM)观察炭黑的特性。将60只9周龄雄性BALB/c小鼠随机分为两个对照组、7天暴露组和14天暴露组,每组动物数量均为15只。小鼠分别在吸入箱中以(29.33±9.10)mg/m³的浓度,每天暴露6小时,连续暴露7天和14天。在暴露7天和14天后,末次暴露24小时后处死小鼠,对照组小鼠在7天和14天时处死。分离气管、肺、肝、肾和脾组织并称重。采用苏木精-伊红(HE)染色,通过光学显微镜观察肺组织的病理变化。采用免疫组织化学法分析肺组织白细胞介素-8(IL-8)的表达。用透射电子显微镜观察肺组织的超微结构。

结果

炭黑暴露14天后,暴露组肺系数较对照组升高(0.61±0.03 vs 0.79±0.06,t = 6.26,P < 0.01)。脾系数高于对照组(0.39±0.04 vs 0.51±0.06,t = 4.23,P < 0.01)。炭黑组与对照组其他器官系数无显著差异。组织病理学显示,7天和14天组炭黑颗粒沉积于肺泡及肺支气管壁。14天组小鼠肺组织内有炭黑颗粒沉积。14天组出现纤毛损伤、肺泡壁严重损伤、炎性细胞浸润及更多充血。免疫组织化学显示,7天(0.272±0.011)和14天(0.422±0.065)暴露组IL-8水平高于14天对照组(0.188±0.041),F = 31.89,P < 0.01。透射电子显微镜显示,对照组肺组织视野清晰,细胞器完整。暴露组肺组织巨噬细胞溶酶体中出现颗粒,电子密度与炭黑颗粒一致。

结论

动态炭黑颗粒暴露可影响小鼠肺和脾系数,破坏肺形态完整性并诱导炎症反应。

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