2,2’,4,4’-四溴联苯醚对人神经母细胞瘤SH-SY5Y细胞胞内游离钙离子浓度的影响

Effect of 2,2',4,4'-tetrabromodiphenyl Ethers on Intracellular Free Calcium Concentration in Human Neuroblastoma SH-SY5Y Cells

目的研究22,’,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([Ca2+]i)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24 h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1 h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、24 h后[Ca2+]i变化。结果与溶剂对照组比较,5、10μmol/L PBDE-47染毒组细胞存活率明显下降(P<0.05);不同浓度PBDE-47染毒1 h后[Ca2+]i浓度均高于染毒0 h(P<0.05);10μmol/LPBDE-47染毒3、6、12、24 h后[Ca2+]i浓度均升高(P<0.05),5μmol/L PBDE-47染毒3、6、24 h后[Ca2+]i浓度也升高(P<0.05),而1μmol/L PBDE-47仅染毒6 h后[Ca2+]i浓度升高(P<0.05)。结论 PBDE-47染毒可使SH-SY5Y细胞[Ca2+]i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.