Heat shock protein 10 modulated apoptosis induced by manganese in SH-SY5Y cell lines

来源 :中国毒理学会第六届全国毒理学大会 | 被引量 : 0次 | 上传用户:jeffery2010
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  OBJECTIVE To examine cell viability, apoptosis and heat shock protein 10 (Hsp10) mRNA expression changes caused by Mn2+ in SH-SY5Y cells.METHODS SH-SY5Y cells are divided into experimental group and control group: Group A (normal saline, 0.9% NaCl) as control group, Group B (Mn2 + 0.5 mmol· L-1), Group C (Mn2 + 1.0 mmol· L-1), Group D (Mn2 + 2.0 mmol· L-1) ,Group E(Mn2+ 4.0 mmol·L-1) as experimental group.Cells are exposed to Mn2+ for 12 h, and then the cell viability was measured by MTT method.Cell Apoptosis was detected using flow cytometry with AnnexinⅤ-FITC/Propidium iodide (PI) staining.Meanwhile, the mRNA expressions of Hsp10 in SH-SY5Y were measured by real-time RT-PCR.RESULTS After exposed to Mn2 + , the cell viability of group B,C, D and E(94.50%, 90.08%, 83.95% and 79.40% respectively) are obviously decreased compared with group A (P<0.01).The rate of cell apoptosis in group B, C, D, and E increased.According to the results of the AnnexinⅤ-FITC mark, the rate of cell apoptosis in group B, group C, group D, and group E increased 2.58, 4.65, 8.64, and 12.76 times as high as normal saline control group respectively;the rate of cell apoptosis in group B and group C increased significantly (P < 0.05);the rate of cell apoptosis in group D and group E increased clearly significantly (P<0.01).According to the PI tag results, the rate of cell apoptosis in group B, C, D and E increased 1.27, 9.23, 5.38, and 3.19 times as high as normal saline control group respectively;compared with control group, the rate of cell apoptosis in group B increased significantly (P < 0.05);the rate of cell apoptosis in group C, D and E increased clearly significantly (P < 0.01).Real-time RT-PCR shows 0.95, 0.84, 0.63, 0.46 folds increase of Hsp10 mRNA expression in group B, C, D, and E respectively compared with group A.In SHSY5Y cells, the rate of cell apoptosis increased but the Hsp10 mRNA decreased with the increased concentration of Mn2 + The correlation analysis between the rate of cell apoptosis and Hsp10 mRNA showed a negative correlation (r=0.996, P<0.01), suggesting that the Hsp10 expression level decreased and the rate of cell apoptosis increased.CONCLUSION With the increased concentration of Mn2+ , Hsp10 mRNA expression and cell viability of SH-SY5Y cells declined, but the rate of cell apoptosis increased.And with the higher concentration of Mn2+ , the Hsp10 mRNA concentration and the rate of cell apoptosis had a negative correlation, suggesting that cell oxidative stress and DNA mutations are the causes making the decrease of Hsp10 mRNA content, cell apoptosis and damage to nerve cells.The experimental results are almost consistent with other research results.
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