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目的 :观察不同浓度锌对锌转运体 1 (zinc transporter1 ,Zn T- 1 ) m RNA和金属硫蛋白 (metallothionein,MT) 1、2m RNA表达的影响 ,旨在探讨神经元锌内稳态的可能机制。 方法 :采用锥虫蓝染色法检测不同浓度锌 (0、5 0、1 0 0、1 2 5、1 5 0、1 75、2 0 0 μm ol/ L)对原代培养海马神经元存活率的影响 ;用实时荧光定量 RT- PCR法分别检测不同浓度锌 (0、5 0、75、1 0 0、1 2 5、1 5 0 μmol/ L)对 Zn T- 1、MT1、MT2的 m RNA表达的影响 ,检测 1 0 0 μm ol/ L 锌暴露不同时间点 (0、2、4、6、8h)对 Zn T- 1、MT1、MT2的 m RNA表达的影响。 结果 :培养基锌浓度大于 1 2 5 μmol/ L 时 ,海马神经元存活率明显下降 ;锌浓度大于 75μm ol/ L 时 ,Zn T- 1 m RNA表达成线性上升 (P<0 .0 5 ) ,MT m RNA表达同样明显上升 (P<0 .0 5 ) ,但在 1 0 0 μm ol/ L 后进入平台期 ;在 1 0 0 μm ol/ L 锌浓度条件下 ,Zn T- 1 m RNA表达在 2 h内达到峰值 ,MT m RNA在 6 h达到其峰值。结论 :虽然神经元可通过增加 Zn T- 1和 MT的表达来应对高锌冲击 ,但两者的保护作用不同 ,Zn T- 1外排作用发生在前且具有持续性 ,MT的络合作用发生在后且会饱和
OBJECTIVE: To observe the effects of different concentrations of zinc on mRNA expression of zinc transporter1 (Zn T-1) m RNA and metallothionein (MT) 1,2 m RNA in order to explore the possibility of zinc homeostasis in neurons mechanism. Methods: The survival rate of primary cultured hippocampal neurons was detected by trypan blue staining with different concentrations of zinc (0, 0, 100, 100, 125, 150, 75 and 200 micromol / L) The effects of different concentrations of zinc (0, 5 0, 75, 100, 125, 150 μmol / L) on the activities of Zn T- 1, MT1 and MT2 m 1, MT1 and MT2 at different time points (0, 2, 4, 6, 8h) were detected. Results: The survival rate of hippocampal neurons was significantly decreased when the concentration of zinc in culture medium was more than 125 micromol / L, and the expression of Zn-1 m RNA increased linearly when the concentration of zinc was higher than 75 micromol / L (P <0.05) , MT m RNA expression also increased significantly (P <0.05), but reached the plateau after 100 μmol / L; at Zn concentration of 100 μmol / L, Zn T-1 m RNA The expression peaked within 2 h, and MT m RNA reached its peak at 6 h. CONCLUSION: Although neurons can respond to high zinc shock by increasing the expression of Zn T-1 and MT, the protective effects of the two are different. The exocytosis of Zn T-1 occurs predominantly and persistently, and the complexation of MT Occurs after and will be saturated