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Technologies and experimental approaches in the NIH Botanical Research Centers

机译:美国国立卫生研究院植物学研究中心的技术和实验方法

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摘要

There are many similarities between research on combinatorial chemistry and natural products and research on dietary supplements and botanicals in the NIH Botanical Research Centers. The technologies in the centers are similar to those used by other NIH-sponsored investigators. All centers rigorously examine the authenticity of botanical dietary supplements and determine the composition and concentrations of the phytochemicals therein, most often by liquid chromatography–mass spectrometry. Several of the centers specialize in fractionation and high-throughput evaluation to identify the individual bioactive agent or a combination of agents. Some centers are using DNA microarray analyses to determine the effects of botanicals on gene transcription with the goal of uncovering the important biochemical pathways they regulate. Other centers focus on bioavailability and uptake, distribution, metabolism, and excretion of the phytochemicals as for all xenobiotics. Because phytochemicals are often complex molecules, synthesis of isotopically labeled forms is carried out by plant cells in culture, followed by careful fractionation. These labeled phytochemicals allow the use of accelerator mass spectrometry to trace the tissue distribution of 14C-labeled proanthocyanidins in animal models of disease. State-of-the-art proteomics and mass spectrometry are also used to identify proteins in selected tissues whose expression and posttranslational modification are influenced by botanicals and dietary supplements. In summary, the skills needed to carry out botanical centers’ research are extensive and may exceed those practiced by most NIH investigators.
机译:美国国立卫生研究院植物学研究中心的组合化学和天然产物研究与膳食补充剂和植物药研究之间有许多相似之处。这些中心的技术与其他由NIH赞助的研究人员所使用的技术相似。所有中心都严格检查植物性膳食补充剂的真伪,并通常通过液相色谱-质谱法确定其中的植物化学成分和浓度。一些中心专门进行分级分离和高通量评估,以鉴定单个生物活性剂或活性剂组合。一些中心正在使用DNA微阵列分析来确定植物药对基因转录的影响,以期揭示其调控的重要生化途径。与所有异种生物一样,其他研究中心也侧重于生物利用度以及植物化学物质的吸收,分布,代谢和排泄。由于植物化学物质通常是复杂的分子,因此同位素标记形式的合成是由培养中的植物细胞进行的,然后进行仔细的分级分离。这些标记的植物化学物质允许使用加速器质谱法来追踪疾病动物模型中 14 C标记的原花色素的组织分布。最新的蛋白质组学和质谱技术还用于鉴定所选组织中的蛋白质,这些蛋白质的表达和翻译后修饰受植物药和膳食补充剂的影响。总而言之,进行植物中心研究所需的技能是广泛的,并且可能超过大多数NIH研究人员所实践的技能。

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