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Effects of Hawthorn Crataegus Plant Extracts on Endothelial Cells and Cardiomyocytes

Effects of Hawthorn Crataegus Plant Extracts on Endothelial Cells and Cardiomyocytes. By Linda Phan Mentor: Dr. Theresa Filtz. Congestive Heart Failure. :disorder where the heart loses the ability to pump blood efficiently. 4.5 million patients in US 400,000 diagnosed each year

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Effects of Hawthorn Crataegus Plant Extracts on Endothelial Cells and Cardiomyocytes

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  1. Effects of Hawthorn Crataegus Plant Extracts on Endothelial Cells and Cardiomyocytes By Linda Phan Mentor: Dr. Theresa Filtz

  2. Congestive Heart Failure :disorder where the heart loses the ability to pump blood efficiently • 4.5 million patients in US • 400,000 diagnosed each year Source: National Institute of Health

  3. Hawthorn Crataegus laevigata • Extracts = Botanical medicine • Alleviates symptoms of congestive heart failure • Increases exercise tolerance • Increases blood flow out of the heart • Affects blood pressure Active ingredients?

  4. Congestive Heart Failure • Causes: • Hypertension = high blood pressure • Hypertrophy = thickening of tissue, caused by increased cell size

  5. My Summer…Methods Development • Endothelial Cells- Project #1 • Hypertension = high blood pressure • Cardiomyocytes - Project #2. w/Dr. Shannon Long

  6. Project #1: Endothelial Cells Endothelial Cells Nitric Oxide (NO) Relaxes Smooth Muscle Relaxed muscle disables contraction = dilation

  7. How does NO relax smooth muscle? NO  Inactivates contraction Dephosphorylated Mechanism of smooth muscle relaxation

  8. Does Hawthorn extract affect NO release in endothelial cells? • Quantify NO release using assays 1. Develop assays able to produce consistent results 2. Then observe Hawthorn’s effects EA.hy926 cells in culture From the Human Umbilical Vein

  9. Nitric Oxide Production Mechanism of NO production in an endothelial cell

  10. Radiotracer Assay’s Mechanism 14C-Arginine  14C-Citrulline • Measure 14C-Citrulline product • Correlate with NO production

  11. Does the protocol work? A23197 – calcium ionophore, increases intracellular [Ca2+] by increasing cell permeability

  12. Nitric Oxide Production Mechanism of NO production in an endothelial cell

  13. Edited diagram by Cayman Chemical

  14. Results

  15. Results

  16. What’s next? • Achieve consistency. • How reliable is Dowex resin? • Is resin holding the unused C-14-Arginine?

  17. Dowex Resin • Ion-exchange • Resin overloaded?

  18. Thin Layer ChromatographyComing Soon! Before Dowex After Dowex

  19. My Summer…Methods Development • Endothelial Cells- Project #1 • Cardiomyocytes - Project #2, w/Dr. Shannon Long • Hypertrophy = thickening of tissue, caused by increased cell size

  20. Project #2: Cardiomyocytes • Heart muscle cells • From mice pups • Contracts in media

  21. Contracted vs. Relaxed • Contracted

  22. Contracted vs. Relaxed • Relaxed

  23. Contracted vs. Relaxed • Relaxed

  24. Contracted vs. Relaxed • Contracted

  25. Have already found … Epinephrine Beating cells

  26. Have already found… Epinephrine NON-Beating cells

  27. Hypertrophy : thickening of tissue, caused by increased cell size • 1.“Create” hypertrophy in cardiomyocytes • Phenylephrine 2. Hypertrophy? 3. Treat with Hawthorn on cells with hypertrophy, observe. (coming soon)

  28. Does Phenylephrine induce hypertrophy?

  29. Results

  30. What’s Next? • Collect more data. • If hypertrophy in cells is successful, treat with Hawthorn.

  31. Dr. Filtz’s Long-Term Goals • Define the active components in Hawthorn with cardiovascular activity. • Fractionation of the extracts to identify active components. • Finally, determine proper dosing and toxic effects.

  32. Acknowledgements • Howard Hughes Medical Institute (HHMI) • Undergraduate Research, Innovation, Science, and Creativity (URISC) • Dr. Theresa Filtz • Dr. Shannon Long • The Filtz Lab • Dr. Kevin Ahern Thank you for your time!

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