Nielsen Henrietta M, Veerhuis Robert, Holmqvist Bo, Janciauskiene Sabina
Department of Clinical Chemistry, Pathology, The Alzheimer Centre, VU University Medical Center Amsterdam, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
Glia. 2009 Jul;57(9):978-88. doi: 10.1002/glia.20822.
Clearance of the amyloid-beta peptide (A beta) as a remedy for Alzheimer's disease (AD) is a major target in on-going clinical trials. In vitro studies confirmed that A beta is taken up by rodent astrocytes, but knowledge on human astrocyte-mediated A beta clearance is sparse. Therefore, by means of flow cytometry and confocal laser scanning microscopy (CLSM), we evaluated the binding and internalization of A beta1-42 by primary human fetal astrocytes and adult astrocytes, isolated from nondemented subjects (n = 8) and AD subjects (n = 6). Furthermore, we analyzed whether alpha1-antichymotrypsin (ACT), which is found in amyloid plaques and can influence A beta fibrillogenesis, affects the A beta uptake by human astrocytes. Upon over night exposure of astrocytes to FAM-labeled A beta1-42 (10 microM) preparations, (80.7 +/- 17.7)% fetal and (52.9 +/- 20.9)% adult A beta-positive astrocytes (P = 0.018) were observed. No significant difference was found in A beta1-42 uptake between AD and non-AD astrocytes, and no influence of ApoE genotype on A beta1-42 uptake was observed in any group. There was no difference in the percentage of A beta-positive cells upon exposure to A beta1-42 (10 microM) combined with ACT (1,000:1, 100:1, and 10:1 molar ratio), versus A beta1-42 alone. CLSM revealed binding of A beta1-42 to the cellular surfaces and cellular internalization of smaller A beta1-42 fragments. Under these conditions, there was no increase in cellular release of the proinflammatory chemokine monocyte-chemoattractant protein 1, as compared with nontreated control astrocytes. Thus, primary human astrocytes derived from different sources can bind and internalize A beta1-42, and fetal astrocytes were more efficient in A beta1-42 uptake than adult astrocytes.
清除淀粉样β肽（Aβ）作为治疗阿尔茨海默病（AD）的方法是正在进行的临床试验的主要目标。体外研究证实Aβ可被啮齿动物星形胶质细胞摄取，但关于人类星形胶质细胞介导的Aβ清除的知识却很稀少。因此，我们通过流式细胞术和共聚焦激光扫描显微镜（CLSM），评估了从非痴呆受试者（n = 8）和AD受试者（n = 6）分离的原代人胎儿星形胶质细胞和成人星形胶质细胞对Aβ1-42的结合和内化。此外，我们分析了在淀粉样斑块中发现的、可影响Aβ纤维形成的α1-抗糜蛋白酶（ACT）是否会影响人类星形胶质细胞对Aβ的摄取。在星形胶质细胞过夜暴露于FAM标记的Aβ1-42（10 microM）制剂后，观察到（80.7 +/- 17.7）%的胎儿星形胶质细胞和（52.9 +/- 20.9）%的成人Aβ阳性星形胶质细胞（P = 0.018）。在AD星形胶质细胞和非AD星形胶质细胞之间，未发现Aβ1-42摄取存在显著差异，且在任何组中均未观察到ApoE基因型对Aβ1-42摄取有影响。在暴露于Aβ1-42（10 microM）并结合ACT（摩尔比为1000:
