Wnt-LRP6信号通路激活REST蛋白的表达并发挥神经保护作用

发布时间：2018-03-19 01:30

本文选题：Wnt-LRP信号通路　切入点：PrP-毒性多肽　出处：《中国农业大学学报》2017年06期 　论文类型：期刊论文

【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-HALRP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。
[Abstract]:To investigate the relationship between Wnt- 尾 -catenin signaling pathway with LRP6 receptor as activation switch and neuronal damage induced by PrP106-126 toxic polypeptide, and to study the correlation between Wnt- 尾 -catenin signaling pathway and neuroprotective protein REST. The effects of Wnt-LRP6-REST on neuronal damage induced by toxic polypeptides were further elucidated. Primary neurons were stimulated by the activator of Wnt signaling pathway, the inhibitor DKK1 or PrP106-126 toxic polypeptide. The changes of downstream proteins associated with neuroprotective protein REST and Wnt signaling pathway were detected by Western blot. The co-localization of positive related protein 尾 -catenin in REST and Wnt signaling pathway was observed by laser confocal method. The overexpression plasmids pCMV-C-HALRP and interference plasmids pGPH1/GFP/Neo-LRP6-Rat shRNA-1 and shRNA-2 of LRP6 were constructed, and the constructed plasmids were transfected into primary neurons. The effects of PrP106-126 toxic polypeptides on Wnt signal pathway iconic proteins (尾 -catenin and GSK3 尾) were detected by over-Western blotting. The effects of overexpression or interference of LRP6 on the downstream proteins of REST or Wnt signaling pathway were detected. The effects of LRP6 on nerve fiber injury induced by toxic peptides were observed by immunofluorescence. Wnt- 尾 -catenin signaling pathway activated by LRP6 receptor can positively regulate the expression of neuroprotective protein REST. LRP6 plays a key role in signal regulation. Moreover, LRP6 can alleviate the neuronal injury induced by PrP106-126 toxic polypeptide. The overexpression of LRP6 increases the activity of neurons and plays a neuroprotective role.
【作者单位】：中国农业大学动物医学院/国家动物海绵状脑病实验室;军事医学科学院实验动物中心;福建农林大学动物科学学院;
【基金】：“十二五”国家科技支撑计划(2015BAI07B02)
【分类号】：R741

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