纳米氧化硅与聚丙烯酸甲酯-纳米氧化硅对大鼠急性肺脏毒性及其机制研究

Acute lung toxicity of nanosilica and polyacrylate-nanosilica and related mechanisms in rats

目的通过纳米氧化硅和聚丙烯酸甲酯-纳米氧化硅染毒大鼠模型,对纳米氧化硅和聚丙烯酸甲酯-纳米氧化硅的急性肺脏毒性及其相关机制进行研究。方法将健康雄性SPF级Wistar大鼠60只随机分为6组,每组10只,即24 h和72 h对照组(0.9%氯化钠溶液气管内滴注后24 h和72 h)、纳米氧化硅24 h组和72 h组(纳米氧化硅溶胶气管内滴注后24 h和72 h)、聚丙烯酸甲酯-纳米氧化硅复合乳液24 h组和72 h组(聚丙烯酸甲酯-纳米氧化硅复合乳液气管内滴注后24 h和72 h)。分别于染毒24 h和72 h后分离大鼠肺脏和胸膜进行HE染色,进行血管内皮生长因子C(VEGF-C)、血管内皮生长因子受体3(VEGFR-3)、转化生长因子β1(TGF-β1)兔多克隆抗体免疫组织化学染色和VEGF-C、VEGFR-3、淋巴管内皮透明质酸受体1(LYVE-1)兔多克隆抗体Western blot实验。结果纳米氧化硅组和聚丙烯酸甲酯-纳米氧化硅组大鼠肺泡间隔增宽、伴炎细胞聚集,胸膜间皮细胞增生,呈乳头状结构。VEGF-C、VEGFR-3和TGF-β1兔多克隆抗体免疫组织化学染色显示,纳米氧化硅组和聚丙烯酸甲酯-纳米氧化硅组大鼠肺脏可见大量组织细胞、炎细胞和淋巴管内皮细胞胞浆染色阳性,胸膜表面较多间皮细胞胞浆染色阳性;Western blot实验显示,纳米氧化硅组和聚丙烯酸甲酯-纳米氧化硅组大鼠VEGF-C、VEGFR-3和LYVE-1蛋白在肺脏中的表达明显增强,差异有统计学意义(P<0.05)。结论纳米氧化硅和聚丙烯酸甲酯-纳米氧化硅染毒可致大鼠出现肺脏损伤,可能与VEGF-C、VEGFR-3、TGF-β1和LYVE-1的高表达有关。

Objective To understand the acute lung toxicity and related mechanisms of nanosilica and polyacrylate-nanosilica in rats.Methods Healthy male Wistar rats(n=60)were randomly divided into six groups,ten in each,four exposure groups(24 h and 72 h)and two corresponding saline control groups.The animals were exposed to either nanosilica,or a polyacrylatenanosilica composite emulsion via intratracheal instillation respectively.After the treatments,the lung tissue and the pleural tissue were taken in each group for the pathological examination.The expression of VEGF-C,VEGFR-3 and TGF-β1 in lung and pleural were determined by using immunohistochemistry.The expression of VEGF-C,VEGFR-3 and LYVE-1 in lung were determined by using Western blot.Results The histopathological examination of lung and pleura in both exposure groups showed thickened alveolar septum,infiltration with inflammatory cells,pleural mesothelium cells hyperplasia.The immunohistochemistry examination of lung and pleura in both exposure groups showed VEGF-C,VEGFR-3 and TGF-β1 positive expression in tissue cells,inflammatory cells,lymphatic endothelial cells and pleural mesothelium cells.The Western blot examination of lung in both exposure groups showed VEGF-C,VEGFR-3 and LYVE-1 positive expression compared with control groups(P<0.05).Conclusion There are lung injuries in the rats after exposure to nanosilica and polyacrylate-nanosilica.The positive expression of VEGF-C,VEGFR-3,TGF-β1 and LYVE-1 is probably the related toxicity mechanisms of nanosilica and polyacrylate-nanosilica.