Uncovering multiple molecular targets for caffeine using a drug target validation strategy combining A 2A receptor knockout mice with microarray profiling.
Uncovering multiple molecular targets for caffeine using a drug target validation strategy combining A 2A receptor knockout mice with microarray profiling.
复制标题
DOI：
10.1152/physiolgenomics.90353.2008
复制
发表时间：
2009-05
影响因子：
4.6
通讯作者：
Liqun Yu;J. Coelho;Xiaoling Zhang;Yutao Fu;Abigail Tillman;U. Karaoz;B. Fredholm;Z. Weng;Jiang-fan Chen
中科院分区：
文献类型：
--
作者：
Liqun Yu;J. Coelho;Xiaoling Zhang;Yutao Fu;Abigail Tillman;U. Karaoz;B. Fredholm;Z. Weng;Jiang-fan Chen
咖啡因是最广泛使用的精神活性物质，在大脑中具有复杂的药理作用。在这项研究中，我们采用了一种新的药物靶点验证策略，使用组合的A（2A）受体（A（2A）R）敲除（KO）和微阵列分析来揭示咖啡因的多个分子靶点。与通过A（2A）R基因缺失或通过将咖啡因给予A（2A）R KO小鼠相比，咖啡因（10 mg/kg）在WT小鼠中引起纹状体基因表达的独特谱。因此，A（2A）Rs是必需的，但不足以引起纹状体基因表达的咖啡因（10 mg/kg）。咖啡因（50 mg/kg）诱导了具有三组不同纹状体基因的复杂表达模式：1）一个子集与A（2A）R基因缺失引起的那些重叠; 2）在WT和A（2A）R KO小鼠中由咖啡因引起的第二个子集; 3）仅在A（2A）R KO小鼠中由咖啡因引起的第三个子集。此外，由磷酸二酯酶（PDE）抑制剂咯利普兰和GABA（A）受体拮抗剂荷包牡丹素引起的纹状体基因集与A（2A）R KO小鼠给予咖啡因（50 mg/kg）引起的纹状体基因的不同子集重叠。最后，基因集富集分析表明，脂肪细胞分化/胰岛素信号在低剂量和高剂量咖啡因引起的纹状体基因集中高度富集。这些不同的纹状体基因群体及其相应的多个分子靶标的鉴定，包括A（2A）R、非A（2A）R（可能是A（1）R和与PDE和GABA（A）R相关的通路）及其相互作用，以及受低剂量和高剂量咖啡因影响的细胞通路，提供了对咖啡因在大脑中的急性药理学作用的分子见解。
Caffeine is the most widely consumed psychoactive substance and has complex pharmacological actions in brain. In this study, we employed a novel drug target validation strategy to uncover the multiple molecular targets of caffeine using combined A(2A) receptor (A(2A)R) knockouts (KO) and microarray profiling. Caffeine (10 mg/kg) elicited a distinct profile of striatal gene expression in WT mice compared with that by A(2A)R gene deletion or by administering caffeine into A(2A)R KO mice. Thus, A(2A)Rs are required but not sufficient to elicit the striatal gene expression by caffeine (10 mg/kg). Caffeine (50 mg/kg) induced complex expression patterns with three distinct sets of striatal genes: 1) one subset overlapped with those elicited by genetic deletion of A(2A)Rs; 2) the second subset elicited by caffeine in WT as well as A(2A)R KO mice; and 3) the third subset elicited by caffeine only in A(2A)R KO mice. Furthermore, striatal gene sets elicited by the phosphodiesterase (PDE) inhibitor rolipram and the GABA(A) receptor antagonist bicucullin, overlapped with the distinct subsets of striatal genes elicited by caffeine (50 mg/kg) administered to A(2A)R KO mice. Finally, Gene Set Enrichment Analysis reveals that adipocyte differentiation/insulin signaling is highly enriched in the striatal gene sets elicited by both low and high doses of caffeine. The identification of these distinct striatal gene populations and their corresponding multiple molecular targets, including A(2A)R, non-A(2A)R (possibly A(1)Rs and pathways associated with PDE and GABA(A)R) and their interactions, and the cellular pathways affected by low and high doses of caffeine, provides molecular insights into the acute pharmacological effects of caffeine in the brain.