| 倪雯雯,谭小芳,于丰祥,宫 平,陈 功,侯丽娜,李 娟.Cu2+-Aβ 复合物与Aβ 单体诱导神经元 H2O2 释放作用的比较研究[J].现代生物医学进展英文版,2018,(8):1401-1406. |
| Cu2+-Aβ 复合物与Aβ 单体诱导神经元 H2O2 释放作用的比较研究 |
| Comparison of the Effects of Cu2+-Aβ Complex and Aβ Monomers on Neuronal Release of H2O2 |
| Received:December 15, 2017 Revised:January 09, 2018 |
| DOI：10.13241/j.cnki.pmb.2018.08.001 |
| 中文关键词: Cu2+-Aβ复合物 Aβ 神经元 活性氧 阿尔茨海默病 |
| 英文关键词: Cu2+-Aβ complex Aβ Neuron Reactive oxygen species Alzheimer's Disease |
| 基金项目:国家自然科学基金项目（81373417，81503044，30973538）；上海市"科技创新行动计划"生物医药领域科技支撑项目（14431901800） |
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| 中文摘要: |
| 摘要 目的：比较不同摩尔比Cu2+-Aβ复合物与Aβ单体诱导神经元H2O2释放作用的差异。方法：制备不同摩尔比（0.1-5）的Cu2+-Aβ复合物，通过检测硫磺素T (Thioflavin T，ThT) 荧光强度考察Cu2+对Aβ纤丝形成的影响。利用原代培养的大鼠海马神经元细胞，分别以不同摩尔比Cu2+-Aβ复合物，不同浓度Cu2+-Aβ复合物（摩尔比为1），以及Aβ单体和Cu2+处理细胞，检测培养上清中的H2O2含量；分离线粒体，分别检测不同浓度Cu2+-Aβ复合物（摩尔比为1），以及Aβ单体和Cu2+处理后H2O2的释放；观察不同摩尔比Cu2+-Aβ复合物，不同浓度Cu2+-Aβ复合物（摩尔比为1），以及Aβ单体和Cu2+对神经元细胞活力的影响。结果：（1）ThT 荧光试验结果表明，Cu2+ 与Aβ（10 μM）摩尔比为1~5范围内可明显抑制Aβ纤丝形成。（2）Cu2+-Aβ复合物（摩尔比为1~5；Aβ浓度为10 μM）以及摩尔比为1的Cu2+-Aβ复合物（Aβ浓度分别为5，10 μM）可显著诱导神经元释放H2O2；另外，摩尔比为1时，Cu2+-Aβ复合物还可诱导神经元线粒体内H2O2释放；上述作用均强于Aβ单体或Cu2+。（3）Cu2+-Aβ复合物（摩尔比为1~5）可显著降低神经元细胞活力，该作用强于Aβ单体或Cu2+ 。结论：与Aβ单体相比，Cu2+-Aβ复合物诱发神经元细胞及其线粒体释放H2O2作用更强，并诱发更为明显的神经元毒性。提示Cu22++与Aβ之间的配位结合可能增强其引发活性氧释放以及神经元毒性反应；Cu2+-Aβ复合物引发的活性氧可能主要来自线粒体。 |
| 英文摘要: |
| ABSTRACT Objective: To compare the effects of Cu2+-Aβ complex at different molar ratios with Aβ monomers on neuronal release of H2O2. Methods: Cu2+-Aβ complex at different molar ratios (0.1-5) were prepared. Thioflavin T (ThT)-based fluorometric assay was used to examine the effect of Cu2+ on Aβ fibril formation. The primary hippocampal neurons were treated with Cu2+-Aβ complex of different ratios, Cu2+-Aβ complex of different concentrations (with a molar ratio of 1), Aβ monomers and Cu2+, respectively. H2O2 content in the culture medium was measured. The mitochondria were separated and treated with different concentrations of Cu2+-Aβ complex (with a molar ratio of 1), Aβ monomers and Cu2+, respectively. H2O2 from mitochondria were also measured. Neuronal viability was detected using Cell Counting Kit-8 (CCK-8) after the neurons were treated with Cu2+-Aβ complex of different ratios, Cu2+-Aβ complex of different concentrations (with a molar ratio of 1), Aβ monomers and Cu2+. Results: (1) ThT-based fluorometric assay demonstrated that Cu2+ inhibited Aβ fibril formation when the molar ratio of Cu2+to Aβ (10 μM) was within 1~5. (2) Cu2+-Aβ complex (the molar ratio was within 1~5; the concentration of Aβ was 10 μM) and Cu2+-Aβ complex with molar ratio of 1 (the concentrations of A were 5, 10 μM) significantly induced neuron releasing of H2O2; In addition, when the molar ratio was 1, Cu2+-Aβ complex also induced neuronal mitochondria releasing of H2O2; All the above mentioned effects were stronger than that of Aβ monomer or Cu2+. (3) Cu2+-Aβ complex (with a molar ratios within 1~5) significantly reduced the viability of neuron cells, which was stronger than that of Aβ monomers or Cu2+. Conclusion: Compared with Aβ monomers, Cu2+-Aβ complex elicited a more robust H2O2 release from neuronal cells and their mitochondria. Moreover, it induced a more overt neuronal cell death. Our observations suggest that Cu2+-Aβ interactions may enhance the ability of Aβ to induce reactive oxygen species release and neuronal toxicity; Cu2+-Aβ complex-evokeded reactive oxygen species may primarily come from the mitochondria. |
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