矽尘致肺泡巨噬细胞产生O_2^的原位显示技术

IN SITU DEMONSTRATION OF O_2~ ON ALVEOLAR MACROPHAGES INDUCED BY SILICIOUS DUSTS

建立无机有毒粉尘颗粒致肺泡巨噬细胞 (alveolar macrophages,AMs)产生超氧阴离子 (O- ·2 )的原位显示技术。以硝基四氮唑蓝 (Nitro Blue Tetrazolium,NBT)为捕捉剂 ,于产生 O- ·2 的原位形成紫蓝色的双甲月替沉淀。O- ·2 的阳性反应呈紫蓝色沉淀 ,分布于 AMs质膜及与吞噬颗粒交界处。结果表明 ,无机有毒粉尘颗粒致 AMs产生的 O- ·2 能够以细胞化学方法于原位显示 ;以 O- ·2 反应阳性为标志 ,AMs为一个异质性群体。这一技术比自由基总体测定技术更能直观具体的显示 AMs与粉尘的作用关系 ,是一项实用的研究技术。

The aim of this study is to set up a morphological technique for in situ demonstration of superoxide anion (O -· 2) in alveolar macrophages (AMs) induced by silicious dusts Using nitro blue tetrozolium (NBT) as a captor, a diformazan precipitate in situ of O -· 2 was formed The positive reaction of O -· 2 was expressed as purple bluished deposits of diformazan, and localized on the cellular margin (cellular plasma) and the borderline between plasma and the ingested particles It has been concluded that (1) O -· 2 produced by alveolar macrophages (AMs) induced by silicious dusts can be in situ manifested by means of cytochemistry technique (2) There is heterogeneity in AMs depending on O -· 2 as a marker In situ evaluation of O -· 2 oxygen free radicals is superior to other techniques in a sense It is more convenient to study directly the real situation of the relationship between the production of oxygen free radicals and the cell peroxide damage induced by silicious dusts \;