Guo Q, Robinson N, Mattson M P
Sanders-Brown Research Center on Aging and Department of Anatomy and Neurobiology, University of Kentucky, Lexington, Kentucky 40536, USA.
J Biol Chem. 1998 May 15;273(20):12341-51. doi: 10.1074/jbc.273.20.12341.
Mutations in the presenilin-1 (PS-1) gene account for approximately 50% of the cases of autosomal dominant, early onset, inherited forms of Alzheimer's disease (AD). PS-1 is an integral membrane protein expressed in neurons and is localized primarily in the endoplasmic reticulum (ER). PS-1 mutations may promote neuronal degeneration by altering the processing of the beta-amyloid precursor protein (APP) and/or by engaging apoptotic pathways. Alternative processing of APP in AD may increase production of neurotoxic amyloid beta-peptide (Abeta) and reduce production of the neuroprotective alpha-secretase-derived form of APP (sAPPalpha). In differentiated PC12 cells expressing an AD-linked PS-1 mutation (L286V), sAPPalpha activated the transcription factor NF-kappaB and prevented apoptosis induced by Abeta. Treatment of cells with kappaB decoy DNA blocked the antiapoptotic action of sAPPalpha, demonstrating the requirement for NF-kappaB activation in the cytoprotective action of sAPPalpha. Cells expressing mutant PS-1 exhibited an aberrant pattern of NF-kappaB activity following exposure to Abeta, which was characterized by enhanced early activation of NF-kappaB followed by a prolonged depression of activity. Blockade of NF-kappaB activity in cells expressing mutant PS-1 by kappaB decoy DNA was associated with enhanced Abeta-induced increases of [Ca2+]i and mitochondrial dysfunction. Treatment of cells with sAPPalpha stabilized [Ca2+]i and mitochondrial function and suppressed oxidative stress by a mechanism involving activation of NF-kappaB. Blockade of ER calcium release prevented (and stimulation of ER calcium release by thapsigargin induced) apoptosis in cells expressing mutant PS-1, suggesting a pivotal role for ER calcium release in the proapoptotic action of mutant PS-1. Finally, a role for NF-kappaB in preventing apoptosis induced by ER calcium release was demonstrated by data showing that sAPPalpha prevents thapsigargin-induced apoptosis, an effect blocked by kappaB decoy DNA. We conclude that sAPPalpha stabilizes cellular calcium homeostasis and protects neural cells against the proapoptotic action of mutant PS-1 by a mechanism involving activation of NF-kappaB. The data further suggest that PS-1 mutations result in aberrant NF-kappaB regulation that may render neurons vulnerable to apoptosis.
早老素-1(PS-1)基因突变约占常染色体显性遗传、早发性、遗传性阿尔茨海默病(AD)病例的50%。PS-1是一种表达于神经元的整合膜蛋白,主要定位于内质网(ER)。PS-1突变可能通过改变β-淀粉样前体蛋白(APP)的加工过程和/或通过启动凋亡途径来促进神经元变性。AD中APP的异常加工可能会增加神经毒性淀粉样β肽(Aβ)的产生,并减少具有神经保护作用的α-分泌酶衍生形式的APP(sAPPα)的产生。在表达与AD相关的PS-1突变(L286V)的分化PC12细胞中,sAPPα激活转录因子NF-κB并阻止Aβ诱导的细胞凋亡。用κB诱饵DNA处理细胞可阻断sAPPα的抗凋亡作用,表明NF-κB激活在sAPPα的细胞保护作用中是必需的。暴露于Aβ后,表达突变型PS-1的细胞表现出异常的NF-κB活性模式,其特征是NF-κB早期激活增强,随后活性长期降低。用κB诱饵DNA阻断表达突变型PS-1的细胞中的NF-κB活性与Aβ诱导的细胞内钙离子浓度([Ca2+]i)升高和线粒体功能障碍增强有关。用sAPPα处理细胞可稳定[Ca2+]i和线粒体功能,并通过涉及激活NF-κB的机制抑制氧化应激。阻断内质网钙释放可防止(而毒胡萝卜素刺激内质网钙释放则诱导)表达突变型PS-1的细胞发生凋亡,提示内质网钙释放在突变型PS-1的促凋亡作用中起关键作用。最后,数据表明sAPPα可防止毒胡萝卜素诱导的细胞凋亡,这一作用被κB诱饵DNA阻断,从而证明了NF-κB在防止内质网钙释放诱导的细胞凋亡中的作用。我们得出结论,sAPPα通过涉及激活NF-κB的机制稳定细胞钙稳态,并保护神经细胞免受突变型PS-1的促凋亡作用。数据进一步表明,PS-1突变导致异常的NF-κB调节,这可能使神经元易受凋亡影响。