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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 the intracellular free calcium ion of human neuroblastoma SH-SY5Y cells [Ca2 +] i) concentration. Methods SH-SY5Y cells in logarithmic growth phase were treated with PBDE-47 solution at concentrations of 0 (solvent control), 1,5 and 10 μmol / L, respectively. The survival rate of SH-SY5Y cells after PBDE-47 in vitro was detected by MTT assay. Fluorescence probe Fluo-3 / AM was used to label SH-SY5Y cells cultured in vitro, and the change of intracellular calcium fluorescence intensity was observed within 1 h under laser scanning confocal microscope. The changes of [Ca2 +] i after 3, 6, 12 and 24 h of PBDE-47 were detected by flow cytometry. Results Compared with the solvent control group, the cell viability of PBDE-47 cells treated with 5 and 10 μmol / L of PBDE-47 significantly decreased (P <0.05). The concentration of [Ca 2+] i in PBDE-47 cells exposed to different concentrations of PBDE- (P <0.05). The concentration of [Ca2 +] i was increased at 10, 6, 12 and 24 h after exposure to 10 μmol / L PDDE-47 (P <0.05) The concentration of [Ca2 +] i also increased (P <0.05), while the concentration of [Ca2 +] i increased in 1μmol / L PBDE-47 treated group (P <0.05) CONCLUSION PBDE-47 can induce early and sustained increase of [Ca2 +] i concentration in SH-SY5Y cells, suggesting that the injury of SH-SY5Y cells induced by PBDE-47 may be related to the change of intracellular calcium homeostasis.