Synergy in Immunomodulation by Spilanthes and Echinacea

描述（由申请人提供）：在过去的十年中，草药的使用在北美地区迅速普及。这些药物是植物成分的复杂混合物，据信这些成分相互作用协同引起生物反应。然而，我们对植物药物生物活性中协同作用的具体机制知之甚少。本研究的目的是研究紫锥菊和Spilanthes这两种据称具有免疫增强活性的植物的化学成分之间的协同作用。这项研究意义重大，因为它有望提供两种药物免疫活性的机制信息，这两种药物在美国和欧洲被广泛用作免疫增强剂。开展这些研究的基本原理是，了解溢菊和紫锥菊提取物与免疫系统相互作用的机制，有望有助于开发利用植物药物治疗或预防与免疫相关的疾病的新策略。本研究的具体目的是确定溢出菊中哪些成分具有免疫调节活性，并评估这些成分相互之间以及与紫锥菊中那些成分协同作用调节免疫活性的潜力。在开展本研究的过程中，将验证香菊中烷基酰胺相互之间以及与香菊和紫锥菊中其他成分基团协同作用以调节免疫活性的假设。这一假设是在初步研究的基础上提出的，这些植物的乙醇提取物富含烷基酰胺，在体外调节淋巴细胞和单核细胞/巨噬细胞的活性。为了验证这一假设，我们将从经过属和种验证的紫锥菊（Echinacea purpurea）和溢草（Spilanthes acmella）植物材料中制备极性不同的植物提取物。这些提取物的化学成分将用液相色谱联用质谱法测定。提取物和分离的成分将检测对人类淋巴细胞和单核/巨噬细胞细胞系细胞活性的影响。将从特别有活性的提取物中纯化单个化合物，并将其活性与复合提取物的活性进行比较。使用肝素化的全血将对特别有效的提取物和分离成分进行测定。结果将使用复杂的协同模型进行分析，以确定具有免疫调节潜力的特定成分组合。这些实验的大部分将在UNCG的化学系进行，该系的本科生由38%的少数民族和60%的女性组成，本科生密切参与研究。该提案是一个多学科的合作努力，研究人员在免疫学和分析化学的专业知识。

DESCRIPTION (provided by applicant): Over the past decade, the use of herbal remedies has skyrocketed in popularity among North Americans. Such remedies are complex mixtures of plant constituents, which are believed to interact synergistically to elicit biological responses. However, very little is known about the specific mechanisms of synergy involved in the biological activities of plant medicines. The objective of the proposed research is to investigate synergistic interactions among the chemical constituents of Echinacea and Spilanthes, two plants with purported immune-enhancing activities. This research is significant because it is expected to provide mechanistic information about the immunological activities of two medicines that are widely employed throughout the US and Europe as immune enhancers. The rationale for carrying out these studies is that an understanding of the mechanisms by which Spilanthes and Echinacea extracts interact with the immune system is expected to aid in the development of novel strategies for the use of plant medicines to treat or prevent diseases with immune related causes. The specific aims of this study are to determine which constituents from Spilanthes have immunomodulatory activity, and to assess the potential that these constituents interact synergistically with each other and with those of Echinacea to modulate immune activity. In carrying out this research, the hypothesis will be tested that alkylamides from Spilanthes interact synergistically with each other and with other constituent groups from Spilanthes and Echinacea to modulate immune activity. This hypothesis has been formulated on the basis of preliminary studies showing that ethanolic extracts of these plants, which are high in alkylamides, modulate the activity of lymphocyte and monocyte/macrophage cells in vitro. To test this hypothesis, plant extracts of varying polarity will be prepared from Echinacea purpurea and Spilanthes acmella plant material that has been verified in terms of genus and species. The chemical composition of these extracts will be determined using liquid chromatography coupled to mass spectrometry. Extracts and isolated constituents will be assayed for effects on activity of cells in human lymphocyte and monocyte/macrophage cell lines. Individual compounds will be purified from particularly active extracts, and their activities will be compared to that of complex extracts. Assays using heparinized whole blood will be performed for particularly active extracts and isolated constituents. The results will be analyzed using a sophisticated synergy model to identify specific constituents combinations with immunomodulatory potential. The bulk of these experiments will be carried out in the department of chemistry at UNCG, where undergraduate enrollment consists of 38% minority and 60% women, and undergraduate students are intimately involved in research. This proposal is a multi-disciplinary, collaborative effort between investigators with expertise in immunology and analytical chemistry.