Microelectrode array biochip: Tool for in vitro drug screening based on the detection of a drug effect on dopamine release from PC12 cells
Microelectrode array biochip: Tool for in vitro drug screening based on the detection of a drug effect on dopamine release from PC12 cells
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DOI：
10.1021/ac060018d
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发表时间：
2006-09-15
影响因子：
7.4
通讯作者：
Sheu, Fwu-Shan
中科院分区：
文献类型：
--
作者：
Cui, Hui-Fang;Ye, Jian-Shan;Sheu, Fwu-Shan
基于微电极阵列（MEA）生物芯片，建立了一种新颖而简单的药物效应检测技术，包括囊泡单胺转运体抑制剂利血平、多巴胺前体左旋多巴和多巴胺转运体抑制剂诺米芬新对多巴胺能PC 12细胞多巴胺释放的影响。在多次注射KCl溶液到附着在MEA生物芯片上的PC 12细胞的培养物中后，通过生物芯片微电极以时间和电流计记录K+刺激的多巴胺释放。本研究定义了记录的电流谱中的两个参数：第一次注射KCl的峰值电流（Max（1））和第四次注射KCl后的稳定电流（St（4））。左旋多巴和利血平的作用在统计学上有显著性差异，通过比较药物中第二次检测的Max 1和St 4与未药物治疗的对照组。将第一次检测中的Max 1和St 4的值归一化为1。相比之下，通过将药物中的第一次检测中的St 4与Max 1的比率与对照的那些进行比较，检测到诺米芬新的统计学显著效果。左旋多巴/利血平和诺米芬辛之间使用不同分析方法测量的原因在于这些药物对PC 12细胞的作用机制不同。开发的新分析方法使用相同的检测装置和参数，并且药物效果的数据分析变得简单。因此，该方法可以提供用于多巴胺相关精神障碍的高通量体外药物筛选方法。
Novel, yet simple detection techniques of drug effect, including the effect of a vesicular monoamine transporter inhibitor ( reserpine), a dopamine precursor (L-dopa), and a dopamine transporter inhibitor ( nomifensine), on dopamine release from dopaminergic PC12 cells were developed based on a microelectrode array (MEA) biochip. Upon multi-injections of KCl solution into the culture of PC12 cells attached on a MEA biochip, the K+-stimulated dopamine release was temporally and amperometrically recorded by biochip microelectrodes. Two parameters in the recorded amperometric spectra were defined in this study: the peak current of the first KCl injection (Max(1)), and the steady current after the fourth KCl injection (St(4)). Statistically significant effects of L-dopa and reserpine were demonstrated by comparing both Max1 and St4 of the second detections in drugs with those of the control without drug treatment. The values of both Max1 and St4 in the first detections were normalized as 1. In contrast, the statistically significant effect of nomifensine was detected by comparing the ratios of St4 to Max1 in the first detections in drug with those of the control. The reason for using different analytical methods for measurements between L-dopa/reserpine and nomifensine lies in the different mechanisms of action on PC12 cells among these drugs. The novel analytical methods developed use the same detection setup and parameters, and the data analysis for the effect of drugs becomes simple. The methods hence may provide a high-throughput in vitro drug screening approach for dopamine-related psychiatric disorders.