Abstract

1. Abstract Echinacea is a common medicinal plant due to its properties of being antiviral and anti-inflammatory. It has been widely used to treat sore throats, coughs, headaches and as a pain reliever. The objective of the current research is to study cytotoxicity of Echinacea ethanol extract fractions and lipopolysaccharides (LPS) on RAW 264.7 mouse macrophage. Fractions Echinacea ethanol extracts and LPS were applied to RAW 264.7 macrophages at the maximum concentrations used in bioactivity assays. After a twenty-four hour incubation, celltiter reagent was added to cell culture with fresh media to measure cell viability. Cell viability of different treatments was recorded as percentage of the vehicle control treatment and ANOVA test was employed for statistical analysis. No cytotoxicity was observed with treatments. The observed anti-inflammatory activity of Echinacea fractions was not a result of reduced cell viability. Results E. Paradoxa fractions Conclusion In conclusion, the results from the cytotoxicity assay showed that the Echinacea paradoxa fractions were not toxic to the cells and even when the fractions were induced with LPS. Nonetheless, LPS did have a major increase of the control percent due to the dehydrogenase activity that took place. Also, the control, ursolic acid, demonstrated cytotoxicity in all the experiments Cytotoxicity of Echinacea Ethanol Fractions and Lipopolysaccharides (LPS) on Raw 264.7 Mouse Macrophages.Leslie R. Nelson1, Xiaozhu Zhang2 , Nan Huang3 , and Diane F. Birt31. University of New Mexico 2. Molecular, Cellular and Developmental Biology Program 3. Department of Food Science and Human Nutrition • Acknowledgments • I like to thank the following for their support and sponsorship: • George Washington Carver Summer Internship Program • National Science Foundation • Dr. Carolyn Lawrence • GWC staff • GWC interns • Center for Integrated Animal Genomics • Center for Research on Botanical Dietary Supplements Fractions 3, 4, 6, and Quercetin Results • Background • Echinacea is a herbal supplement that is native to North America and Southern Canada. It was traditionally used by Native Americans to cure common colds, sore throats, treat infected wounds and headaches. It is known to have antiviral, antioxidant, and anti-inflammatory properties and can be taken orally in the form of tea. • Common names: Latin names: • American Coneflower Echinacea augustifolia • Hedgehog Echinacea paradoxa • Kansas Snake Root Echinacea atrorubens • Black Susan Echinacea pallida • Coneflower Echinacea laevigata • Echinacea purpurea • Echinacea sanguinea • Echinacea simulata • Echinacea tenneseessnsis Fractions 3, 4, 6 and Quercetin with LPS Induction Results Reference “Echinacea.” National Center for Complementary and Alternative Medicine. 17 February 2009. 13 July 2009. http://nccam.nih.gov/health/echinacea/ataglance.htm Booden, Michelle. Raw 264.7 macrophage cells “BALB/c.” Wikipedia: The Free Encyclopedia. 19 June 2009. 13 July 2009. http://en.wikipedia.org/wiki/BALB/c “Cytotoxicity.” The Free Dictionary. 13 July 2009. http://www.thefreedictionary.com/cytotoxicity “Echinacea.” Drug Information Online. 4 July 2009. 13 July 2009. http://www.drugs.com/mtm/echinacea.html “Echinacea.” Wikipedia: The Free Encyclopedia. 13 July 2009. 17 July 2009. http://en.wikipedia.org/wiki/Echinacea Method The method used for this specific research was Cytotoxicity Assay. It is a three day experiment that determines if the Echinacea fractions and controls are toxic to the Raw 264.7 mouse macrophage cells. • Day Two • Spent media is discarded and fresh media is added. • Fresh media is given . • Fractions and controls are induced into the cell wells. • Incubate for 24 hours. • Day Three • Odd media is removed and new media is added. • Cell titer reagent is added to cells. • Incubate for 3 hours and 15 minutes. • Transfer cells to a 96 well plate. • Have the plate read using KCJunior • Day One • Cells are transferred from flask to a 48 well plate. • Dead and live cells are counted. • Incubate for 24 hours. LPS Results