STUDY OF SAFETY ASSESSMENT OF NEW MINERAL FIBERS USING MAKERS IN LUNG FIBROSIS-RELATED BIOLOGICAL DIAGNOSIS

研究了肺泡巨噬细胞释放肺纤维化相关细胞因子的新方法在石棉替代品等矿物纤维安全性评价中的应用价值。(1)人肺泡巨噬细胞释放的tnf - α、il - β、IL-8、mip -1 α和GRO均增加。(2)人成纤维细胞与石棉共培养可加速人肺泡巨噬细胞中il -1 β和GRO的升高。(3)在人肺泡巨噬细胞中发现的基础上，我们测试了MMMF、三种晶须、UICC crocidolite和tremolite三种陶瓷纤维和微玻璃棉对小鼠巨噬细胞样细胞释放il -1 β和tnf - α的影响。(4)在细胞毒性试验中，1 ~ 10mg /ml浓度下的这些纤维均无作用，而100mg ~ 1mg/ml浓度下的Sic晶须仅表现出高细胞毒性。(5) K-Ti-O-晶须和TiO晶须效果相同，陶瓷纤维组、微玻璃棉和SiC晶须在MTT测试中表现出较高的灵敏度。(6)虽然与UICC crocidolite相比，MMMF或晶须增加了细胞培养基中的tnf - α，但陶瓷纤维的作用大于晶须。(7) MMMF细胞内il -1 β的含量与UICC crocidolite细胞内相同或略有升高，而须含量低于UICC crocidolite细胞内。这些结果表明，测量包括肺纤维化相关细胞因子在内的tnf - α和il - β可能有助于石棉替代品的安全性评估。但这些石棉代用品的细胞毒性试验、MTT试验、tnf - α和il - β试验结果不同，需要与体内和体外实验进行比较。

We studied about usefulness of new method which measured lung fibrosis-related cytokines released from alveolar macropahges in safety evaluation of mineral fibers including asbestos substitute.(1) TNF-alpha, IL-beta, IL-8, MIP-1alpha and GRO released from human alveolar macrophages were increased by UICC amosite, crocidolite and chrysotile.(2) Co-culture of human fibrobrasts accelerated the increases of IL-1beta and GRO in human alvelar macrophages with asbestos.(3) On the basis of finding in human alveolar macrophage, we tested effects of three kind of ceramic fibers and micro glass wool including MMMF,three kind of whiskers, UICC crocidolite and tremolite on IL-1beta and TNF-alpha released from mouse macrophage-like cells.(4) In cytotoxicity test, all of these fibers in 1-10mug/ml showed no-effect and Sic whisker of 100mug-1mg/ml only revealed high cytotoxicity.(5) K-Ti-O- and TiO whiskers showed same effect, ceramic fiber group, micro glass wool and SiC whisker showed high sensitivity in MTT test when compared with UICC crocidolite.(6) Although TNF-alpha in cell culture medium increased by MMMF or whisker compared with UICC crocidolite, effect of ceramic fiber was bigger than that of whiskers.(7) Intracellular content of IL-1beta showed same level or a little increase in MMMF,and whisker revealed lower than that of UICC crocidolite.From these results, it suggested that measurements of TNF-alpha and IL-beta including lung fibrosis-related cytokines might be useful in safety evaluation of asbestos substitute. However, different finding between cytotoxicity test, MTT test, TNF-alpha and IL-beta required the comparison with in vivo and in vitro experiments in these asbestos substitutes.