无机砷及其代谢产物诱导体外培养人皮肤成纤维细胞的增殖作用

Effects of proliferation on primary cultured human skin flbroblast induced by ingorganic arsenics and metabolites

目的探讨无机砷（iAsⅢ）、单甲基胂酸（DMAⅤ）、二甲基胂酸（MMAⅢ）对人皮肤成纤维细胞增殖作用的影响。方法原代培养的人皮肤成纤维细胞,通过直接细胞计数法和噻唑蓝（MTT）还原法检测iAsⅢ、DMAⅤ、MMAⅢ对人皮肤成纤维细胞生长的影响。结果荧光倒置显微镜下观察原代1、1~13代人皮肤成纤维细胞形态;与对照组比较,人皮肤成纤维细胞经不同浓度的iAsⅢ、DMAⅤ、MMAⅢ诱导后,吸光度值均有改变,存在剂量-效应关系（P〈0.05）。0.5~5.0μmol/L浓度的iAsⅢ、DMAⅤ、MMAⅢ对人皮肤成纤维细胞有明显的增殖作用,而10~15μmol/L浓度的iAsⅢ、DMAⅤ、MMAⅢ产生明显的毒性作用,毒性比较MMAⅢ〉iAsⅢ〉DMAⅤ。MMAⅢ对人皮肤成纤维细胞有明显的增殖作用,且峰值出现在0~1μmol/L。结论低浓度无机砷及其代谢产物刺激人皮肤成纤维细胞增殖,高浓度具有细胞毒性。

Objective To investigate the effects of arsenic on primary cuitured skin flbroblast induced by iAsⅢ,DMAⅤ,MMAⅢ.Methods Cell number counting,hiazolyl blue（MTT） reduction assay,transmission inverted microscope were performed in primary cultured skin flbroblast and iAsⅢ,DMAⅤ,MMAⅢ were used as tester inorganic arsenic.Results Primary and F11~F13 cultured skin flbroblast with inverted microscope were oberved;With different concentration of iAsⅢ,DMAⅤ,MMAⅢ（0.5~5.0 μmol/L）,the skin fibroblasts were markedly stimulated,and a dose-effect relationship were observed.At concentration of 10~15 μmol/L,however,iAsⅢ,DMAⅤ,MMAⅢ induced injury of skin fibroblasts were observed.Toxicity comparison of skin flbroblast MMAⅢiAsⅢ DMAⅤ.At concentration of 0.5~5.0 μmol/L,MMAⅢ induced proliferation of skin fibroblasts was observed,the crest value was between 0 μmol/L and 1 μmol/L.Conclusion The proliferation of the skin flbroblast line is regulated in a double-phase manner,it may play a critical role in the development of skin injuries caused by inorganic arsenics and the mechanism of DNA injury in primary cultured human skin cells need to be studied further.