Effects of digestion on composition and bioavailability of compounds from Ashwagandha: an in vitro study

摘要 睡莲，在阿育吠陀医学中一直以其药用价值而闻名。近年来， 在西方文化中，睡莲产品的消费量飙升，目前这些产品正在西方文化中销售。 通过零售店作为膳食补充剂，统称为Ashwagandha。NCCIH赞助的 BENFRA植物膳食补充剂研究中心(U19 AT010829)目前正在研究 阿什瓦甘达，以确定其对年龄相关的神经退化的影响。与药物不同 在公开分发之前有一个广泛的评估过程，关于如何 阿什瓦甘达提取物中存在的复杂化学混合物一旦被食用并通过即可发挥作用。 通过消化环境。这项建议是创新的，因为它将评估 首次使用模拟胃肠道的体外实验室模型提取阿什瓦甘达提取物 环境。该项目还将评估哪些原始或修改的成分能够通过肠道 大多数化合物通过被动扩散进入肠道循环。 这项研究的中心假设是阿什瓦甘达衍生代谢物的组成进入 循环量与原提取物有明显不同。这一假设将通过 具体目标如下：1.确定胃肠道环境对小鼠代谢产物的影响 通过将提取物暴露在模拟胃肠液和酶中而提取的阿什瓦甘达提取物；以及2.鉴定 阿什瓦甘达原液和消化孵育液中代谢物的渗透性 肠膜的模型。我们将使用这些体外模型系统的组合，适当 控制和足够的重复，以便进行强有力的统计分析，使这些初步调查标准化。超强的 萃取物的分析将采用加压液相色谱-高分辨质谱联用技术， 培养物和扩散室介质，以评估化学结构和相对丰度的变化 阿什瓦甘达代谢物首次出现在这些复杂的基质中。这些研究的结果将提供 深入了解胃肠道消化后更现实的Ashwagandha代谢情况，然后可以 使用已建立的临床前检测方法进行进一步的合作研究。本研究的结果还可以 重点介绍目前尚未进行生物活性研究的可渗透代谢物。从长远来看，这 该项目将建立一种强大的分析策略，可广泛用于评估其他药物的口服生物利用度 体外环境中的植物补充剂，在时间、劳动力和资源密集型体内研究之前。

Abstract Withania somnifera, has long been known in Ayurvedic medicine for its medicinal properties. In recent years, consumption of W. somnifera products has skyrocketed in western cultures where they are now marketed through retail outlets as dietary supplements, under the common name, Ashwagandha. The NCCIH-sponsored BENFRA Botanical Dietary Supplements Research Center (U19 AT010829) is currently researching Ashwagandha, to determine its effects on age-related neurological deterioration. Unlike pharmaceutical drugs which have an extensive evaluation process prior to public distribution, very little is known regarding how the complex mixture of chemicals present in Ashwagandha extracts behaves once it is consumed and passes through the digestive environment. This proposal is innovative because it will evaluate metabolic stability of Ashwagandha extracts for the first time using in vitro laboratory models simulating the gastrointestinal environment. The project will also assess which original or modified components are able to pass the intestinal membrane via passive diffusion, the mechanism by which most compounds enter the circulation from the gut. The central hypothesis of this research is that the composition of Ashwagandha derived metabolites entering circulation is markedly different from that of the original extract. This hypothesis will be tested through the following specific aims: 1. Determine effects of gastrointestinal environments on the metabolite profiles of Ashwagandha extracts by exposing extracts to simulated gastro-intestinal fluids and enzymes; and 2. Identify permeabilities of metabolites from Ashwagandha original extracts and digestive incubations using a synthetic model of the intestinal membrane. We will use a combination of these in vitro model systems, appropriate controls, and sufficient replicates for robust statistical analyses to standardize these initial investigations. Ultra- pressure liquid chromatography coupled with high resolution mass spectrometry will be used to analyze extracts, incubates, and diffusion chamber media, to evaluate changes in chemical structures and relative abundances of Ashwagandha metabolites in these complex matrices for the first time. Results from these studies will provide insight into a more realistic Ashwagandha metabolic profile post-gastrointestinal digestion that can then be further studied collaboratively using established pre-clinical assays. Results of the present study may also highlight permeable metabolites of interest not currently under investigation for bioactivity. In the long term, this project will establish a robust analytical strategy that can be used broadly to assess oral bioavailability of other botanical supplements in an in vitro setting, prior to time, labor, and resource intensive in vivo studies.