Effects of HNE-modification induced by Abeta on neprilysin expression and activity in SH-SY5Y cells.
Effects of HNE-modification induced by Abeta on neprilysin expression and activity in SH-SY5Y cells.
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DOI：
10.1111/j.1471-4159.2008.05855.x
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发表时间：
2009-02
影响因子：
4.7
通讯作者：
Wang DS
中科院分区：
文献类型：
--
作者：
Wang R;Wang S;Malter JS;Wang DS
β-淀粉样蛋白（Aβ）的脑内蓄积是阿尔茨海默病（AD）的一贯特征，并可能促进AD的发展。除了失调的生产，越来越多的实验证据表明，减少catalysts也在Aβ积累中发挥重要作用。我们先前已经表明，脑啡肽酶（NEP），体内切割Aβ的主要蛋白酶，在AD患者的脑中被4-羟基-壬烯醛（HNE）加合物修饰。为了确定这些变化是否影响Aβ，用HNE或Aβ处理SH-SY 5 Y细胞，然后测定NEP mRNA、蛋白水平、HNE加合的NEP、NEP活性和分泌的Aβ水平。细胞内NEP开发HNE加合物后24小时的HNE治疗所确定的免疫沉淀，免疫印迹和双重免疫荧光染色。HNE修饰的NEP显示催化活性降低，这与SH-SY 5 Y和H4 APP 695 wt细胞中Aβ1-40的升高有关。细胞与Aβ1-42孵育也诱导HNE加合NEP。在一个明显的代偿反应中，Aβ处理的细胞显示NEP mRNA和蛋白表达增加。尽管NEP蛋白升高，但活性与NEP蛋白水平相比显著降低。目前的研究表明，NEP可以通过HNE内收而失活，这至少部分与Aβ切割减少和Aβ积累增加有关。
The cerebral accumulation of β-amyloid (Aβ) is a consistent feature of and likely contributor to the development of Alzheimer’s Disease (AD). In addition to dysregulated production, increasing experimental evidence suggests reduced catabolism also plays an important role in Aβ accumulation. We have previously shown that neprilysin (NEP), the major protease which cleaves Aβ in vivo, is modified by 4-hydroxy-nonenal (HNE) adducts in the brain of AD patients. In order to determine if these changes affected Aβ, SH-SY5Y cells were treated with HNE or Aβ, and then NEP mRNA, protein levels, HNE adducted NEP, NEP activity and secreted Aβ levels were determined. Intracellular NEP developed HNE adducts after 24 h of HNE treatment as determined by immunoprecipitation, immunoblotting and double immunofluorescence staining. HNE-modified NEP showed decreased catalytic activity, which was associated with elevations in Aβ1-40 in SH-SY5Y and H4 APP695wt cells. Incubation of cells with Aβ1-42 also induced HNE adduction of NEP. In an apparent compensatory response, Aβ treated cells showed increased NEP mRNA and protein expression. Despite elevations in NEP protein, the activity was significantly lower compared to the NEP protein level. The present study demonstrates that NEP can be inactivated by HNE-adduction, which is associated with, at least partly, reduced Aβ cleavage and enhanced Aβ accumulation.
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