青石棉处理的肺泡巨噬细胞上清液对人胚肺成纤维细胞的作用

Role of Supernatants treated by Crocidolite on Human Embryonic Pulmonary Fibroblasts

为了探讨青石棉对肺损伤的机理 ,用兔肺泡巨噬细胞(AM)和人胚肺成纤维细胞 (HEPF)组成体外模型 ,采用MTT颜色反映法测定青石棉处理兔AM 2 4h后的培养上清液刺激HEPF细胞增殖情况 ,同时用标准石英作为阳性对照 ,二氧化钛作为阴性对照 ,以不用粉尘处理的AM培养上清液作为正常对照。结果发现青石棉能明显刺激HEPF增殖 ,呈剂量 效应关系 (P <0 0 1)。青石棉浓度在 10 0 μg/ml时 ,青石棉组HEPF的净增殖率达 6 0 91% ,是正常对照组的 3 48倍 ,高于阳性对照和阴性对照 10 0 μg/ml青石棉处理兔AM的培养上清液的不同稀释度对HEPF增殖的影响不同 ,随稀释度的加大 ,其促细胞增殖的能力越弱 ,以 1∶2稀释度作用最强 ,明显高于阳性对照和阴性对照 (P <0 0 1)。提示青石棉处理肺泡巨噬细胞的培养上清液中存在有促进肺成纤维细胞增殖的生物活性物质 。

In order to explore the mechanism of crocidolite injured lung,a in vitro model was formed with rabbit alveolar macrophage(AM)and human embryonic pulmonary fibroblasts(HEPF),the supernatant of AMs treated by crocidolite for 24 hours was prepared,and the proliferation of HEPF stimulated by supernatants of crocidolite treated AMs was measured by MTT color response method.At the same time,setting up positive control(SiO 2),negative control(TiO 2),normal control(AM) and blank control(B.Control).The results showed that the supernatants of crocidolite treated rabbit AM significantly stimulated the proliferation of HEPF with dose effect relationship(P<0 01).The net proliferative rate of HEPF in 100μg/ml crocidolite group reached 60 91%,higher than that of SiO 2 or TiO 2 group,and it was 3 48 times that of AM group.The higher dilution ratio was the supernatants of AM treated by 100μg/ml dusts,the less proliferation was HEPF,its dose effect relationship was significant(P<0 01),and there was stronger ability to stimulate the proliferation of HEPF at 1∶2 dilution ratio of supernatants in different group than that of any other dilution ratio.It was suggested that there exist some bioactive factors in the supematants of crocidolite treated AM to stimulate the proliferation of HEPF.If the synthesis and secretion of those bioactive factors could be inhibited,it would make it possible providing the new way of prevention and treatment for asbestos related diseases.