缺锌致海马神经细胞损伤的表观遗传机制初探

发布时间：2018-03-11 03:34

本文选题：缺锌　切入点：海马神经细胞　出处：《营养学报》2017年04期 　论文类型：期刊论文

【摘要】：目的观察锌缺乏致原代培养的大鼠海马神经细胞损伤中DNA甲基化及组蛋白去乙酰化相关酶的变化,对缺锌致海马神经细胞损伤的表观遗传机制进行初探。方法将原代培养的大鼠海马神经细胞随机分为对照(control);缺锌[四吡啶甲基乙二胺,N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine,TPEN];补锌5和50μmol/L(TPEN+5μmol/L Zn SO_4,TPEN+50μmol/L Zn SO_4)4组,除对照组不加任何处理外,其余3组分别在培养液中加入2μmol/L TPEN,补锌组则对应加入5和50μmol/L Zn SO_4,作用24h。MTT法检测细胞存活率;免疫酶学法检测HDAC活性;RT-PCR法检测DNA甲基转移酶(DNMT3a、DNMT3b)及组蛋白去乙酰化酶HDACs(HDAC1、HDAC2、HDAC3)的m RNA表达水平。结果与对照组比较,(1)缺锌组海马神经存活率明显降低(P0.05);5μmol/L补锌和50μmol/L补锌均可显著抑制TPEN诱导的细胞存活率降低(P0.05)。(2)缺锌组海马神经细胞内DNMT1 m RNA的表达明显上调,DNMT3a m RNA的表达明显下调(P0.05),而5μmol/L补锌组或50μmol/L补锌组均能够显著抑制TPEN诱导的DNMT1 m RNA和DNMT3a m RNA表达异常(P0.05)。与对照组比较,缺锌组海马神经细胞内DNMT3b m RNA的表达无明显变化(P0.05),与对照组比较,50μmol/L补锌后,DNMT3b m RNA表达明显上调(P0.05)。(3)缺锌组海马神经细胞内HDAC活性及HDAC2的m RNA表达明显升高(P0.05),HDAC1及HDAC3 m RNA的表达无明显变化(P0.05);补锌能显著抑制TPEN诱导的HDAC活性及HDAC2 m RNA表达异常(P0.05)。结论细胞内缺锌诱导海马神经细胞损伤,造成DNA甲基化和组蛋白去乙酰化修饰的改变。
[Abstract]:Objective to observe the changes of DNA methylation and histone deacetylation related enzymes in primary cultured rat hippocampal neurons injured by zinc deficiency. Methods the primary cultured rat hippocampal neurons were randomly divided into three groups: control control group; zinc deficiency group [tetramethylenediamineurium pyridylmethyldiethylenediamine]; zinc supplementation (5 渭 mol/L(TPEN) and 50 渭 mol/L(TPEN 5 渭 mol/L ZnSO4 / TPEN 50 渭 mol/L Zn SO_4)4 group. In addition to the control group without any treatment, 2 渭 mol/L TPEN was added into the culture medium in the other three groups, and the zinc supplementation group was supplemented with 5 and 50 渭 mol/L ZnSSO4 respectively. The survival rate of the cells was measured by 24 h. The expression of m RNA in DNA methyltransferase (DNA) and histone deacetylase (HDAC1HDAC2HDAC3) was detected by RT-PCR and immunoenzymatic assay. Results compared with the control group, the survival rate of hippocampal nerve in the zinc-deficient group was significantly lower than that in the control group. The survival rate of hippocampal nerve in the zinc-deficient group was significantly lower than that in the zinc-deficient group. The expression of DNMT1 m RNA in hippocampal neurons of zinc-deficient group was significantly up-regulated and down-regulated P0.05 RNA expression, while 5 渭 mol/L zinc supplementation group or 50 渭 mol/L zinc supplementation group could significantly inhibit TPEN induced DNMT1. The expression of m RNA and DNMT3a m RNA was abnormal (P 0.05). There was no significant change in the expression of DNMT3b m RNA in hippocampal neurons in zinc-deficient group. Compared with the control group, the expression of DNMT3b m RNA increased significantly after zinc supplementation with 50 渭 mol/L. The activity of HDAC and the expression of m RNA of HDAC2 in hippocampal neurons of zinc-deficient group significantly increased HDAC1 and HDAC3 m. Zinc supplementation can significantly inhibit the HDAC activity induced by TPEN and the abnormal expression of HDAC2 m RNA. Conclusion Zinc deficiency can induce hippocampal nerve cell injury. Changes in DNA methylation and histone deacetylation were caused.
【作者单位】：军事医学科学院卫生学环境医学研究所;成都军区疾病预防控制中心;
【基金】：天津市应用基础与前沿技术研究计划(No.15JCYBJC27100) 国家自然科学基金(No.30901185)
【分类号】：R151

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