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Indication of nerve growth factor binding components from herbal extracts by HerboChip: a platform for drug screening on a chip

机译:HerboChip指示来自草药提取物的神经生长因子结合成分:在芯片上进行药物筛选的平台

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Background HerboChip is an array of different fractions deriving from herbal extracts. This study aimed to identify effective components from Chinese medicine (CM) that interact with nerve growth factor (NGF) as a target using HerboChip. Methods Fifty types of CM that are traditionally used as remedies for emotion imbalance were selected and extracted with 50?% ethanol. Biotinylated-NGF was hybridized with over 300 chips coated with different HPLC-separated fractions from CM extracts and straptavidin-Cy5 was used to identify the NGF-bound fractions. Results Over 300 chips were screened within a week, and 17 positive hits were identified. The interaction of the identified herbal extracts with NGF was confirmed in cultured PC12 cells. Co-application of NGF and herbal extract interfered with NGF-induced expression of neurofilaments, including NF68 and NF200 in cell cultures. Western blot analysis comparing the intensity of phosphorylated cAMP response element-binding protein (CREB) over total CREB showed NGF-induced CREB phosphorylation was modulated by the identified herbal extracts. Five CM herbs showed activating activities on the NGF response and nine CM herbs showed inhibiting activities. Conclusion The current result supported the applicability of HerboChip for screening NGF binding components from herbal extracts.
机译:背景HerboChip是源自草药提取物的一系列不同馏分。这项研究旨在使用HerboChip识别与神经生长因子(NGF)相互作用的中药(CM)有效成分。方法选择50种传统上用于情绪失衡的CM,并用50%乙醇提取。将生物素化的NGF与300多个芯片进行杂交,这些芯片包被了来自CM提取物的不同HPLC分离的级分,并使用抗生物素蛋白Cy5来鉴定结合了NGF的级分。结果一周内筛选了300多个筹码,鉴定出17个阳性结果。在培养的PC12细胞中确认了已鉴定的草药提取物与NGF的相互作用。 NGF和草药提取物的共同应用会干扰NGF诱导的细胞培养物中神经丝(包括NF68和NF200)的表达。蛋白质印迹分析比较了磷酸化的cAMP反应元件结合蛋白(CREB)相对于总CREB的强度,发现NGF诱导的CREB磷酸化受到鉴定的草药提取物的调节。 5种CM草药显示出对NGF应答的激活活性,而9种CM草药显示出抑制活性。结论目前的结果支持了HerboChip在从草药提取物中筛选NGF结合成分的适用性。

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