目的研究2,2’,4,4’-四溴联苯醚(2,2’,4,4’tetrabromodiphenyl ethers,PBDE-47)对人神经母细胞瘤SH—SY5Y细胞胞内游离钙离子(【Ca^2+】,)浓度的影响。方法取对数生长期的SH—SY5Y细胞,分别染毒终浓度为0(溶剂对照)、1、5、10μmol/L的PBDE-47溶液。采用MTT法检测PBDE-47体外染毒24h后SH—SY5Y细胞存活率。利用钙离子荧光探针Fluo-3/AM标记体外培养的SH—SY5Y细胞,在激光扫描共聚焦显微镜下持续动态观测1h内胞内钙离子荧光强度变化。采用流式细胞仪检测PBDE-47染毒3、6、12、24h后[Ca^2+]变化。结果与溶剂对照组比较,5、10μmol/LPBDE-47染毒组细胞存活率明显下降(P〈0.05);不同浓度PBDE-47染毒1h后[Ca^2+]浓度均高于染毒0h(P〈0.05);10μmol/LPBDE-47染毒3、6、12、24h后[Ca^2+]浓度均升高(P〈0.05),5μmol/LPBDE-47染毒3、6、24h后[Ca^2+],浓度也升高(P〈0.05),而1肛mol/LPBDE-47仅染毒6h后[Ca^2+];浓度升高(P〈0.05)。结论PBDE-47染毒可使SH—SY5Y细胞[Ca^2+].浓度发生早期且持续升高,提示PBDE-47导致的SH—SY5Y细胞损伤可能与细胞内钙离子稳态改变有关。
Objective To investigate the effect of 2,2' ,4,4' -tetrabromodiphenyl ethers (PBDE-47) on intracellular free calcium ([Ca^2+]i) concentration in human neuroblastoma SH-SY5Y cells. Methods Exponentially growing cells were treated with PBDE-47 at the doses of 0 (solvent control), 1,5 and 10 μmol/L respectively. MTF method was used to detect the cell viability in SH-SY5Y cells treated for 24 h. Fluo-3/AM, a fluorescent calcium indicator, was used to monitor changes of the intracellular calcium signaling, under the laser scanning confocal microscope the [Ca^2+]; level was consecutively detected for 1 h. After 3,6,12 and 24 h of treatment, the levels of [Ca^2+]; were measured respectively by using flow cytometry. Results Compared with the solvent control,the cell viabilities significantly decreased in the cells treated by 5,10 μmol/L PBDE-47 (P〈0.05). In the cells treated by different concentrations of PBDE-47 for 1 h, the level of [Ca^2+]; was higher than that in 0 h group (P〈0.05). Furthermore,after exposure to 10 μmol/L PBDE-47 for 3,6,12 and 24 h, the levels of [Ca^2+]; increased significandy (P〈0.05) and after exposure to 5 μmol/L PBDE-47 for 3,6 and 24 h, the levels of [Ca^2+]; significantly increased also (P〈0.05), but only at 6 h after exposure to 1 μmol/L PBDE-47,the level of [Ca^2+]; increased significantly (P〈0.05). Conclusion PBDE-47 can induce the level of [Ca^2+]; increase at the early stage in SH-SY5Y cells,and the elevation of [Ca^2+]; is long-lasting,which indicates that calcium homeostasis may be related to the SH-SY5Y cells damage induced by PBDE-47.