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

我们研究了新的方法，测量肺纤维化相关细胞因子释放的肺泡巨噬细胞在矿物纤维，包括石棉替代品的安全性评价的有用性。(1)UICC铁石棉、青石棉和温石棉可增加人肺泡巨噬细胞释放TNF-α、IL-β、IL-8、MIP-1 α和GRO。(2)人成纤维细胞与石棉共培养促进人肺泡巨噬细胞IL-1 β和GRO的增加。(3)在人肺泡巨噬细胞实验的基础上，我们检测了三种陶瓷纤维和微晶玻璃棉（包括MMMF、三种晶须、UICC青石棉和透闪石）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- 10 μ g/ml的SiC纤维均无作用，100 μ g-1 mg/ml的SiC晶须仅表现出较高的细胞毒性。(5)K-Ti-O-晶须和TiO晶须的效果相当，陶瓷纤维组、微晶玻璃棉和SiC晶须的MTT检测灵敏度高于UICC青石棉。(6)与UICC青石棉相比，MMMF和晶须均能增加细胞培养液中TNF-α的含量，但陶瓷纤维的作用大于晶须。(7)MMMF组细胞内IL-1 β含量与UICC组相当或略有增加，晶须含量低于UICC组，提示检测TNF-α、IL-β等肺纤维化相关细胞因子的含量可作为石棉替代品安全性评价的参考指标。然而，细胞毒性试验、MTT试验、TNF-α和IL-β之间的不同发现需要与这些石棉替代物的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.