Department of Medical Chemistry, University of Szeged, Szeged, Hungary.
Brain Res Bull. 2012 Apr 10;87(6):521-5. doi: 10.1016/j.brainresbull.2012.02.005. Epub 2012 Feb 25.
It is difficult task to measure precisely the toxic effect of beta-amyloid (Aβ 1-42) peptides and also the protective effect of novel drug candidates against Aβ-peptides. The widely used MTT-assay in cell lines or primary cell cultures could be insensitive against Aβ-peptides. We describe here an easy and relevant method for testing Aβ 1-42 toxicity on acute hippocampal slices derived from rat. Brain slice viability in different conditions was measured using MTT and LDH assays. The concomitant use of these two assays can give detailed and relevant results on the toxic effect of Aβ 1-42 in oxygen-glucose deprived (OGD) acute brain slice model. Both assays are capable of quantifying tissue viability by measuring optical density (OD). We found that simultaneous application of OGD and Aβ 1-42 treatment induced a more intensive decrease in hippocampal slice viability than their separate effects. The use of MTT and LDH assay for quantifying brain slice viability proved to be an easy ex vivo method for investigating Aβ toxicity. Testing brain slices is more relevant in Alzheimer's Disease research than using in vitro cell cultures, due to maintenance of the three dimensional cellular network, the cell variability and intact cell connections.
准确测量β-淀粉样蛋白（Aβ1-42）肽的毒性作用以及新型药物候选物对 Aβ-肽的保护作用是一项艰巨的任务。广泛应用于细胞系或原代细胞培养物中的 MTT 测定法可能对 Aβ-肽不敏感。我们在这里描述了一种在源自大鼠的急性海马切片上测试 Aβ1-42 毒性的简单而相关的方法。使用 MTT 和 LDH 测定法测量了不同条件下脑切片的活力。这两种测定法的同时使用可以在氧葡萄糖剥夺（OGD）急性脑切片模型中提供关于 Aβ1-42 毒性的详细和相关结果。这两种测定法都能够通过测量光密度（OD）来定量组织活力。我们发现，OGD 和 Aβ1-42 处理的同时应用比它们的单独作用诱导了更强烈的海马切片活力下降。MTT 和 LDH 测定法用于定量脑切片活力被证明是一种用于研究 Aβ 毒性的简单离体方法。与使用体外细胞培养物相比，测试脑切片在阿尔茨海默病研究中更相关，因为它维持了三维细胞网络、细胞变异性和完整的细胞连接。
