摘要
目的:观察不同锌水平对体外应激海马神经元金属硫蛋白(MTs)亚型表达的影响。方法:取新生1dWis-tar大鼠海马组织进行体外神经元培养,无血清培养24h后,分别向培养液中加入皮质酮、Zn2+特异鳌合剂TPEN,使二者的最终浓度均为1×10-5mol/L,然后加入不同浓度的ZnSO4溶液,使Zn2+的最终浓度分别为1×10-5mol/L、1×10-4mol/L和2×10-4mol/L,作用24h后检测培养液中IL-6和NO含量,以蛋白印迹法检测细胞MTs含量,以RT-PCR检测细胞MT-1mRNA和MT-3mRNA的表达水平。结果:在海马神经元培养液中加入TPEN后,MTs的表达出现明显降低,皮质酮刺激也未见其表达升高。在补锌组,MTs的含量均明显增加,其中以10-4mol/LZn2+组的表达量最高。海马神经元MT-1mRNA和MT-3mRNA的表达水平在皮质酮应激组和补锌组均出现明显升高。另外,锌缺乏和皮质酮刺激可使海马神经元培养上清中的IL-6和NO水平均出现明显升高。结论:不同锌水平对应激海马神经元金属硫蛋白及其亚型mRNA的表达具有调控作用,缺锌可降低金属硫蛋白的表达,而补锌可增加金属硫蛋白的表达。
To evaluate the effects of different doses of zinc on the expression of metallothionein isoforms in stressed hippocampal neurons in vitro. Methods： The cell stress model was developed by corticosterone. The cultured hippocampal neurons were assigned to seven groups as follows： control group, zinc deficiency group, and their corresponding stressed groups, as well as three different levels of zinc complementarity groups. Results： In zinc deficiency group, the expressions of metallothionein and MT-1 mRNA, MT-3 mRNA were downregulated. On the other hand, inductions of metallothionein and it＇s mRNAs in stressed zinc complementarity group were increased. In addition, the levels of supematant IL-6 and NO were increased clearly in zinc deficiency group and corticosterone stressed groups. Conclusion： Our results suggest that zinc deficiency may decrease while zinc complementarety increase the expressions of metallothioneins and MT-1 mRNA, MT-3 mRNA in stressed hippccampal neurons in vitro.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2006年第4期415-418,共4页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助课题(30100149)
关键词
应激
金属硫蛋白
锌
皮质酮
stress
metallothioneins
zinc
corticosterone
