A novel assay for high-throughput screening of anti-Alzheimer's disease drugs to determine their efficacy by real-time monitoring of changes in PC12 cell proliferation.
A novel assay for high-throughput screening of anti-Alzheimer's disease drugs to determine their efficacy by real-time monitoring of changes in PC12 cell proliferation.
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DOI：
10.3892/ijmm.2013.1608
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发表时间：
2014-03
影响因子：
5.4
通讯作者：
Wang Q
中科院分区：
文献类型：
--
作者：
Hou XQ;Yan R;Yang C;Zhang L;Su RY;Liu SJ;Zhang SJ;He WQ;Fang SH;Cheng SY;Su ZR;Chen YB;Wang Q
阿尔茨海默病(AD)是一种神经退行性疾病，其特征是在包括大脑皮层和海马体在内的大脑中积累老年斑和神经原纤维缠结形成。目前，AD的一线治疗是应用乙酰胆碱酯酶抑制剂。然而，乙酰胆碱酯酶抑制剂基本上对AD病理的有限方面具有抗症状作用，并与严重的副作用有关。中草药具有多靶点、多途径、多系统的优势，具有开发治疗阿尔茨海默病药物的潜力。近年来，随着中草药研究和体外药理研究的发展，细胞疾病模型已成为筛选中草药潜在疗效的主要方法之一。由于缺乏客观的高通量筛选方法，测试可能的抗阿尔茨海默病药物的疗效和开发新药受到阻碍。目前，药物疗效的评价通常采用四甲基偶氮唑盐比色法，涉及繁琐、主观、低通量的方法。在此，我们提出了一种实时细胞监测设备(XCELLigence)的新应用，该设备可以通过评估淀粉样蛋白β多肽Aβ1-42诱导的PC12细胞凋亡来简单、客观地评估中药化合物的有效成分。我们使用全自动、高通量和实时系统检测细胞的增殖和运动。我们定量测定了不同抗AD药物干预Aβ1-42诱导PC12细胞凋亡的不同发育阶段的实时IC50值，并根据疗效确定了最佳的作用时间段。我们的数据表明，这项技术可能有助于发现和开发新的抗阿尔茨海默病药物。可以利用类似的技术来测量细胞在培养皿中附着和扩散时的电阻抗变化，培养皿上覆盖着覆盖井底大约80%的面积的金微电极阵列。当细胞附着和铺展在电极表面时，导致电阻抗增加9-12。阻抗表现为称为细胞指数的无量纲参数，它与细胞覆盖的组织培养井的总面积成正比。因此，细胞指数可以用来监测细胞的黏附、铺展、形态变异和细胞密度。
Alzheimer’s disease (AD) is a neurodegenerative disease that is characterized by the accumulation of senile plaque and neurofibrilary tangle formation in the brain, including the cerebral cortex and hippocampus. Nowadays, the first-line treatment for AD is the application of acetylcholinesterase inhibitors. However, acetylcholinesterase inhibitors are basically anti-symptomatic for a limited aspect of AD pathology and are associated with serious side-effects. With the advantage of multiple targets, pathways and systems, Chinese herbal compounds hold promising potential for the development of drugs for the treatment of AD. Over the past few years, with the development of Chinese herbal compounds and in vitro pharmacological studies, cell-based disease models are one of the main methods used to screen Chinese herbal compounds for potential efficacy. Testing the efficacy of possible anti-Alzheimer’s disease drugs and the development of new drugs are hindered by the lack of objective high-throughput screening methods. Currently, the assessment of the effects of drugs is usually made by MTT assays, involving laborious, subjective, low-throughput methods. Herein, we suggest a novel application for a real-time cell monitoring device (xCELLigence) that can simply and objectively assess the effective composition of Chinese herbal compounds by assessing amyloid-β peptide Aβ1-42-induced apoptosis in PC12 cells. We detected the proliferation and motility of the cells using a fully automated high-throughput and real-time system. We quantitatively assessed cell motility and determined the real-time IC50 values of various anti-AD drugs that intervene in several developmental stages of Aβ1-42-induced apoptosis in PC12 cells, Then, we identified the optimal time phase by curative efficacy. Our data indicate that this technique may aid in the discovery and development of novel anti-Alzheimer’s disease drugs. It is possible to utilize a similar technique to measure changes in electrical impedance as cells attach and spread in a culture dish covered with a gold microelectrode array that covers approximately 80% of the area on the bottom of a well. As cells attach and spread on the electrode surface, it leads to an increase in electrical impedance of 9–12. The impedance is displayed as a dimensionless parameter termed the cell index, which is directly proportional to the total area of tissue culture well that is covered by the cells. Hence, the cell index can be used to monitor cell adhesion, spreading, morphological variation and cell density.
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影响因子：
3.1
作者：
Ifeoma, Obidike;Samuel, Okhale;Adeola, Salawu Oluwakanyinsola
通讯作者：
Adeola, Salawu Oluwakanyinsola
影响因子：
3.5
作者：
Onoue, S;Ohshima, K;Kashimoto, K
通讯作者：
Kashimoto, K
影响因子：
2.9
作者：
Thorn, DC;Meehan, S;Carver, JA
通讯作者：
Carver, JA
DOI：
10.1073/pnas.88.17.7896
发表时间：
1991-09-01
影响因子：
11.1
作者：
GIAEVER, I;KEESE, CR
通讯作者：
KEESE, CR
影响因子：
3.9
作者：
Li, Xu Ling;Wang, De Sheng;Sun, Man Ji
通讯作者：
Sun, Man Ji