Screening a specific Zn(II)-binding peptide for improving the cognitive decline of Alzheimer's disease in APP/PS1 transgenic mice by inhibiting Zn2 -mediated amyloid protein aggregation and neurotoxicity
Screening a specific Zn(II)-binding peptide for improving the cognitive decline of Alzheimer's disease in APP/PS1 transgenic mice by inhibiting Zn2 -mediated amyloid protein aggregation and neurotoxicity
复制标题
筛选特定的 Zn(II) 结合肽，通过抑制 Zn2 介导的淀粉样蛋白聚集和神经毒性来改善 APP/PS1 转基因小鼠阿尔茨海默病的认知衰退
DOI：
10.1039/c9bm00676a
复制
发表时间：
2019
影响因子：
6.6
通讯作者：
Gao Hui-Ling
中科院分区：
文献类型：
--
作者：
Zhang Xiao-Yu;Zhong Man-Li;Zhao Pu;Zhang Xian-Cheng;Li You;Wang Xu-Liang;Sun Jia;Lan Wang;Sun He-Hong;Wang Zhan-You;Gao Hui-Ling
Zn 2+与阿尔茨海默病（AD）的进展有关，因为淀粉样β蛋白（Aβ）聚集和神经毒性是由锌离子介导的。因此，开发用于抑制和调节金属触发的Aβ聚集的金属螯合剂作为治疗AD的策略受到关注。在这里，我们使用基于噬菌体展示的方法来筛选特异性阻断Zn触发的Aβ聚集的Zn（II）结合肽。用Ni-IDA亲和树脂制备固定化Zn（II）的树脂，通过与噬菌体七肽库的相互作用，筛选出目标Zn（II）。经IDA阴性筛选和Zn（II）阳性筛选4轮后，获得了高特异性的Zn（II）结合酶。通过DNA测序和ELISA检测，共鉴定出15组组氨酸含量较高的锌结合肽。我们选择了一个高度特异性的Zn（II）多肽（序列为H-M-Q-T-N-H-H），并对其螯合Zn 2+和抑制Zn 2+介导的Aβ聚集的能力进行了体外研究。我们将Zn（II）结合肽负载到PEG修饰的壳聚糖纳米颗粒（NPs）上，以提高Zn（II）结合肽的稳定性和生物利用度。负载有Zn（II）结合肽的PEG修饰的壳聚糖NPs（PEG/PZn-CS NPs）降低了稳定过表达APP Swedish突变（N2 aswe）的小鼠神经母细胞瘤（N）2a细胞中的Zn 2+浓度和Aβ分泌。PEG/PZn-CS纳米粒可减轻Zn 2+诱导的神经毒性、氧化应激和细胞凋亡。鼻内给药PEG/PZn-CS NPs可改善APP swe/PS1 d9（APP/PS1）双转基因小鼠的认知能力，减少小鼠脑内Aβ斑块。这项研究表明，锌（II）结合肽及其纳米粒子有希望作为一个潜在的抗AD剂。
Zn2+ has been implicated in the progression of Alzheimer's disease (AD), as amyloid-β protein (Aβ) aggregation and neurotoxicity are mediated by zinc ions. Therefore, development of metal chelators for inhibiting and regulating metal-triggered Aβ aggregation has received attention as a strategy for treating AD. Here, we used an approach based on phage display to screen for a Zn(II)-binding peptide that specifically blocks Zn-triggered Aβ aggregation. A fixed Zn(II) resin was prepared using Ni-IDA affinity resin, and the target Zn(II) was screened by interaction with a heptapeptide phage library. After negative biopanning against IDA and four rounds of positive biopanning against Zn(II), high specificity Zn(II)-binding phages were obtained. Through DNA sequencing and ELISA, 15 sets of Zn(II)-binding peptides with high histidine contents were identified. We chose a highly specific peptide against Zn(II) with the sequence of H-M-Q-T-N-H-H, and its abilities to chelate Zn2+ and inhibit Zn2+-mediated Aβ aggregation were assessed in vitro. We loaded the Zn(II)-binding peptide onto PEG-modified chitosan nanoparticles (NPs) to improve the stability and the bioavailability of the Zn(II) binding peptide. PEG-modified chitosan NPs loaded with Zn(II)-binding peptide (PEG/PZn-CS NPs) reduced Zn2+ concentrations and Aβ secretion in mouse neuroblastoma (N)2a cells stably over-expressing the APP Swedish mutation (N2aswe). Zn2+-Induced neurotoxicity, oxidative stress, and apoptosis were attenuated by PEG/PZn-CS NPs. Intranasal administration of PEG/PZn-CS NPs improved the cognitive ability of APPswe/PS1d9 (APP/PS1) double-transgenic mice and reduced Aβ plaques in the mouse brain. This study indicated that a Zn(II)-binding peptide and its NPs have promise as a potential anti-AD agent.