本文選題：Wnt-LRP信號(hào)通路　切入點(diǎn)：PrP-毒性多肽　出處：《中國(guó)農(nóng)業(yè)大學(xué)學(xué)報(bào)》2017年06期 　論文類型：期刊論文

【摘要】：為檢測(cè)以LRP6受體為激活開關(guān)的Wnt-β-catenin信號(hào)通路與PrP106-126毒性多肽引起的神經(jīng)元損傷之間的相互關(guān)系,并研究該信號(hào)通路與神經(jīng)保護(hù)性蛋白R(shí)EST的相關(guān)性,從而進(jìn)一步闡明Wnt-LRP6-REST對(duì)毒性多肽引起的神經(jīng)元損傷的影響。利用Wnt信號(hào)通路的激活劑Licl、抑制劑DKK1或PrP106-126毒性多肽刺激原代神經(jīng)元,通過免疫印跡檢測(cè)神經(jīng)保護(hù)性蛋白R(shí)EST及Wnt信號(hào)通路下游相關(guān)蛋白的變化情況,通過激光共聚焦觀察REST與Wnt信號(hào)通路正相關(guān)蛋白β-catenin的共定位情況。分別構(gòu)建LRP6的過表達(dá)質(zhì)粒pCMV-C-HALRP和干擾質(zhì)粒pGPH1/GFP/Neo-LRP6-Rat shRNA-1和shRNA-2,將已經(jīng)構(gòu)建好的質(zhì)粒轉(zhuǎn)染進(jìn)入原代神經(jīng)元。通過免疫印跡檢測(cè)PrP106-126毒性多肽對(duì)Wnt信號(hào)通路標(biāo)志性蛋白(β-catenin及GSK3β)的影響,檢測(cè)LRP6的過表達(dá)或干擾對(duì)REST或Wnt信號(hào)通路下游蛋白的影響。通過免疫熒光觀察LRP6對(duì)由毒性多肽誘導(dǎo)的神經(jīng)纖維損傷的影響,用流式細(xì)胞儀檢測(cè)相應(yīng)的細(xì)胞活力。由LRP6受體激活的Wnt-β-catenin信號(hào)通路在一定程度上正向調(diào)控神經(jīng)保護(hù)性蛋白R(shí)EST的表達(dá),LRP6在信號(hào)調(diào)節(jié)的過程中起到了關(guān)鍵作用,并且LRP6對(duì)由PrP106-126毒性多肽引起的神經(jīng)元損傷有緩解作用,LRP6的過表達(dá)增加了神經(jīng)元細(xì)胞的活力,起到了神經(jīng)保護(hù)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.
【作者單位】： 中國(guó)農(nóng)業(yè)大學(xué)動(dòng)物醫(yī)學(xué)院/國(guó)家動(dòng)物海綿狀腦病實(shí)驗(yàn)室;軍事醫(yī)學(xué)科學(xué)院實(shí)驗(yàn)動(dòng)物中心;福建農(nóng)林大學(xué)動(dòng)物科學(xué)學(xué)院;
【基金】：“十二五”國(guó)家科技支撐計(jì)劃(2015BAI07B02)
【分類號(hào)】：R741

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