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| Wnt-LRP6信号通路激活REST蛋白的表达并发挥神经保护作用 |
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宋志琦1, 王进2, 朱婷3, 周向梅1, 杨利峰1, 赵德明1
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| (1.中国农业大学 动物医学院/国家动物海绵状脑病实验室, 北京 100193;2.军事医学科学院 实验动物中心, 北京 100071;3.福建农林大学 动物科学学院, 福州 350002) |
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| 摘要: |
| 为检测以LRP6受体为激活开关的Wnt-β-catenin信号通路与PrP106-126毒性多肽引起的神经元损伤之间的相互关系，并研究该信号通路与神经保护性蛋白REST的相关性，从而进一步阐明Wnt-LRP6-REST对毒性多肽引起的神经元损伤的影响。利用Wnt信号通路的激活剂Licl、抑制剂DKK1或PrP106-126毒性多肽刺激原代神经元，通过免疫印迹检测神经保护性蛋白REST及Wnt信号通路下游相关蛋白的变化情况，通过激光共聚焦观察REST与Wnt信号通路正相关蛋白β-catenin的共定位情况。分别构建LRP6的过表达质粒pCMV-C-HA-LRP和干扰质粒pGPH1/GFP/Neo-LRP6-Rat shRNA-1 和shRNA-2，将已经构建好的质粒转染进入原代神经元。通过免疫印迹检测PrP106-126毒性多肽对Wnt信号通路标志性蛋白（β-catenin及GSK3β）的影响，检测LRP6的过表达或干扰对REST或Wnt信号通路下游蛋白的影响。通过免疫荧光观察LRP6对由毒性多肽诱导的神经纤维损伤的影响，用流式细胞仪检测相应的细胞活力。由LRP6受体激活的Wnt-β-catenin信号通路在一定程度上正向调控神经保护性蛋白REST的表达，LRP6在信号调节的过程中起到了关键作用，并且LRP6对由PrP106-126毒性多肽引起的神经元损伤有缓解作用，LRP6的过表达增加了神经元细胞的活力，起到了神经保护作用。 |
| 关键词: Wnt-LRP6信号通路 PrP106-126毒性多肽 原代神经元 神经保护 |
| DOI：10.11841/j.issn.1007-4333.2017.06.12 |
| 投稿时间：2016-05-23 |
| 基金项目:“十二五”国家科技支撑计划（2015BAI07B02） |
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| Wnt-LRP6 signaling pathway regulates the expression of REST and its neuroprotective function |
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SONG Zhiqi1, WANG Jin2, ZHU Ting3, ZHOU Xiangmei1, YANG Lifeng1, ZHAO Deming1
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| (1.College of Veterinary Medicine/National Animal Transmissible Spongiform Encephalopathy Laboratory, China Agricultural University, Beijing 100193, China;2.Laboratory Animal Center, Academy of Military Medical Science, Beijing 100071, China;3.College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China) |
| Abstract: |
| Aims of this study were to investigate the influence of PrP106-126 on canonical Wnt-LRP6 pathway and the relationship between low-density lipoprotein receptor-related protein 6 (LRP6) and RE1-silencing transcription factor(REST) in PrP106-126-induced neuropathological changes.Wnt-LRP6 inhibitor and activator were used to test the effect of Wnt signaling on REST.By overexpression or knockdown of LRP6,the relationship between LRP6 and REST in PrP106-126-induced neuropathological changes was examined by immunofluorescence and Western blot analysis.The function of LRP6 in PrP106-126-stimulated primary neuronal death was verified by Annexin V-FITC/PI apoptosis assay.The results showed that Wnt signaling partly regulated the expression of REST;LRP6 was critical for the activation of REST and protects neurons from PrP106-126-induced synaptic and neurofibrillary degeneration. In conclusion,the LRP6-Wnt-β-catenin/REST signaling played critical and collaborative roles in neuroprotection.This signaling of neuronal survival regulation could be explored as a viable therapeutic target for prion diseases and associated neurodegenerative diseases. |
| Key words: Wnt-LRP6 signaling pathway PrP106-126 primary cortical neurons neuroprotection |
