Location: Plant Introduction Research
Project Number: 5030-21000-058-03-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Sep 18, 2013
End Date: Sep 15, 2016
In an earlier collaboration between the University of Louisiana Monroe and NCRPIS, we demonstrated stimulation of bone marrow progenitor cell production for the myeloid lineage (GM-CFCs) to be nearly equivalent for a commercial Echinacea supplement and aerial parts of accession E. purpurea PI 649040. The work demonstrated that the herbal supplement activity was due to phytoconstituents, rather than non-plant additives. The newly proposed studies will demonstrate whether potency of PI 649040 is similarly enhanced by lipophilic extraction, as shown for the supplement, and will provide justification for future studies to develop an Echincacea-derived myelostimulatory phytopharmaceutical, in part, by screening NCRPIS germplasm for optimal starting material. We anticipate that an efficacious, plant-derived myelostimulant will have application for treatment of several clinical instances associated with bone marrow failure (BMF), especially acquired BMF as occurs with cancer chemotherapy and myelodysplasia syndromes resulting from occupational exposures to hematotoxicants such as benzene. The goal of this project is a focused assessment of whether the processing step shown to markedly enhance potency of pharmacological activity of botanical supplement Echinacea is effective with the same type of plant material as indicated on the supplement label, when produced from source-identified germplasm curated by the North Central Regional Plant Introduction Station (NCRPIS), Ames, IA.
Aerial plant materials (stems, leaves, and flowers) from Echinacea purpurea PI 649040 will be produced at the USDA-ARS North Central Regional Plant Introduction Station and shipped to the University of Louisiana at Monroe. Upon arrival, the plant materials will be dried, powdered, and macerated with 75% ethanol at room temperature. The ethanol extract will be n-hexane-washed and the extracts lyophilized and purified using high performance thin layer chromatography (HPTLC). Female rats will receive six daily oral doses of 0 – 200 mg/kg of the extract. Twenty-four hours later, bone marrow cells (BMCs) will be plated in 96-well plates in methylcellulose containing colony stimulating factor (CSF2), interleukin-3 (IL-3), and stem cell factor (SCF) (HALO kit, HemoGenix, Colorado Springs, CO). After five days, ATP of bone marrow progenitor cells of myeloid lineage (GM-CFCs) will be measured by a luminescence assay. Concurrent chemical analyses will be done with HPTLC and proton nuclear magnetic resonance spectroscopy (1H-NMR). At conclusion we should have a dataset from E. purpurea PI 649040 for direct comparison to that available from plant material used to prepare Echinacea herbal supplement.