Aliev Gjumrakch, Seyidova Dilara, Neal Maxwell L, Shi Jiong, Lamb Bruce T, Siedlak Sandra L, Vinters Harry V, Head Elizabeth, Perry George, Lamanna Joseph C, Friedland Robert P, Cotman Carl W
Microscopy Research Center, Department of Anatomy, Case Western Reserve University and University Hospitals of Cleveland, 2085 Adelbert Road, Cleveland, OH 44106, USA.
Ann N Y Acad Sci. 2002 Nov;977:45-64. doi: 10.1111/j.1749-6632.2002.tb04798.x.
We have studied the ultrastructural features of vascular lesions and mitochondria in brain vascular wall cells from human AD brain biopsy, human short postmortem brain tissues, and yeast artificial chromosome (YAC) and C57B6/SJL transgenic positive (Tg+) mice overexpressing amyloid beta precursor protein (AbetaPP). In situ hybridization using mitochondrial DNA (mtDNA) probes for human wild type, 5 kb deleted, and mouse mtDNA was performed, along with immunocytochemistry using antibodies against amyloid precursor protein (APP), 8-hydroxy-2'-guanosine (8-OHG), and cytochrome c oxidase (COX). There was a higher degree of amyloid deposition in the vascular walls of the human AD, YAC, and C57B6/SJL Tg (+) mice compared to age-matched controls. In addition, vessels with more severe lesions showed immunopositive staining for APP and possessed large, lipid-laden vacuoles in the cytoplasm of endothelial cells (EC). Significantly more mitochondrial abnormalities were seen in human AD, YAC, and C57B6/SJL Tg (+) mouse microvessels where lesions occurred. In situ hybridization using wild and chimera (5 kb) mtDNA probes revealed positive signals in damaged mitochondria from the vascular endothelium and in perivascular cells of lesioned microvessels close to regions of large amyloid deposition. These features were absent in undamaged regions of human AD tissues, YAC and C57B6/SJL Tg (+) mouse tissues, and in age-matched control subjects. In addition, vessels with atherosclerotic lesions revealed endothelium and perivascular cells possessing clusters of wild and deleted mtDNA positive probes. These mtDNA deletions were accompanied by increased amounts of immunoreactive APP, 8-OHG, and COX in the same cellular compartment. Our observations demonstrate that vascular wall cells, especially their mitochondria, appear to be a central target for oxidative stress-induced damage.
我们研究了取自人类阿尔茨海默病（AD）脑活检组织、人类死后短时间脑组织、以及过表达淀粉样β前体蛋白（AβPP）的酵母人工染色体（YAC）和C57B6/SJL转基因阳性（Tg+）小鼠的脑血管壁细胞中血管病变和线粒体的超微结构特征。使用针对人类野生型、缺失5 kb的线粒体DNA（mtDNA）探针以及小鼠mtDNA进行原位杂交，并使用针对淀粉样前体蛋白（APP）、8-羟基-2'-鸟苷（8-OHG）和细胞色素c氧化酶（COX）的抗体进行免疫细胞化学分析。与年龄匹配的对照组相比，人类AD、YAC和C57B6/SJL Tg（+）小鼠的血管壁中淀粉样沉积程度更高。此外，病变更严重的血管显示APP免疫阳性染色，并且在内皮细胞（EC）的细胞质中存在大的、充满脂质的空泡。在发生病变的人类AD、YAC和C57B6/SJL Tg（+）小鼠微血管中，观察到的线粒体异常明显更多。使用野生型和嵌合（5 kb）mtDNA探针进行原位杂交，在来自血管内皮的受损线粒体以及靠近大量淀粉样沉积区域的病变微血管的血管周围细胞中发现阳性信号。这些特征在人类AD组织、YAC和C57B6/SJL Tg（+）小鼠组织的未受损区域以及年龄匹配的对照受试者中均不存在。此外，具有动脉粥样硬化病变的血管显示内皮细胞和血管周围细胞拥有野生型和缺失型mtDNA阳性探针簇。这些mtDNA缺失伴随着同一细胞区室中免疫反应性APP、8-OHG和COX含量的增加。我们的观察结果表明，血管壁细胞，尤其是它们的线粒体，似乎是氧化应激诱导损伤的主要靶点。
