Immunomodulatory Effects of Glycerolic Echinacea purpurea Stem and Leaf Extract on RAW 264.7 Murine Macrophages
Location
Room 232, Schewel Hall
Access Type
Open Access
Entry Number
82
Start Date
4-5-2023 10:15 AM
End Date
4-5-2023 10:30 AM
College
Lynchburg College of Arts and Sciences
Department
Biomedical Science
Keywords
Echinacea, Echinacea purpurea, glycerine, RAW 264.7, Macrophage, Inflammation, Herbal, Extract, Immunomodulation
Abstract
The eastern and central North American flowering plant Echinacea purpurea (EP) is used by Natives as a traditional remedy for upper respiratory illnesses. Active chemical constituents for EP are caffeic acid derivatives, alkylamides, and polysaccharides. EP extracts within the context of human systems have been suggested to shorten the duration of upper respiratory illness symptoms. The majority of studies suggest an immunomodulatory effect on innate immune responses. This research will test the effects of a 50:50 glycerol:water extraction of EP stem and leaf on LPS-stimulated RAW 264.7 murine macrophages. Results will be compared to previous work using 75:25 ethanolic root and 75:25 ethanolic stem and leaf extract to determine differences in direct effect on RAW 264.7 cells. RAW 264.7 cells plated at 4x10^5 cells/well (500uL DMEM) in a 24-well plate are stimulated with bacterial lipopolysaccharide [LPS] (E. coli O55:B5) for 24 hours. Supernatants are reserved for cytokine ELISA and a sample used to measure nitrites in solution by the Greiss reaction to assess inflammatory responses. Preliminary experiments with extract diluted in DMEM at 15uL/mL (0.75% glycerine), 30uL/mL (1.5% glycerine) and 60uL/mL (3% glycerine) showed inhibitory effect in cells stimulated by 100 ng/mL of LPS. Further experiments will examine lower concentrations of glycerine in extract in the culture environment to elucidate solvents effect on cell culture responses.
Faculty Mentor(s)
Dr. David Freier
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Immunomodulatory Effects of Glycerolic Echinacea purpurea Stem and Leaf Extract on RAW 264.7 Murine Macrophages
Room 232, Schewel Hall
The eastern and central North American flowering plant Echinacea purpurea (EP) is used by Natives as a traditional remedy for upper respiratory illnesses. Active chemical constituents for EP are caffeic acid derivatives, alkylamides, and polysaccharides. EP extracts within the context of human systems have been suggested to shorten the duration of upper respiratory illness symptoms. The majority of studies suggest an immunomodulatory effect on innate immune responses. This research will test the effects of a 50:50 glycerol:water extraction of EP stem and leaf on LPS-stimulated RAW 264.7 murine macrophages. Results will be compared to previous work using 75:25 ethanolic root and 75:25 ethanolic stem and leaf extract to determine differences in direct effect on RAW 264.7 cells. RAW 264.7 cells plated at 4x10^5 cells/well (500uL DMEM) in a 24-well plate are stimulated with bacterial lipopolysaccharide [LPS] (E. coli O55:B5) for 24 hours. Supernatants are reserved for cytokine ELISA and a sample used to measure nitrites in solution by the Greiss reaction to assess inflammatory responses. Preliminary experiments with extract diluted in DMEM at 15uL/mL (0.75% glycerine), 30uL/mL (1.5% glycerine) and 60uL/mL (3% glycerine) showed inhibitory effect in cells stimulated by 100 ng/mL of LPS. Further experiments will examine lower concentrations of glycerine in extract in the culture environment to elucidate solvents effect on cell culture responses.