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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

Effects of Hawthorn Crataegus Plant Extracts on Endothelial Cells and Cardiomyocytes

By Linda Phan

Mentor: Dr. Theresa Filtz

congestive heart failure
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

hawthorn crataegus laevigata
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?

congestive heart failure1
Congestive Heart Failure
  • Causes:
    • Hypertension = high blood pressure
    • Hypertrophy = thickening of tissue, caused by increased cell size
my summer methods development
My Summer…Methods Development
  • Endothelial Cells- Project #1
      • Hypertension = high blood pressure
  • Cardiomyocytes - Project #2.

w/Dr. Shannon Long

project 1 endothelial cells
Project #1: Endothelial Cells

Endothelial Cells

Nitric Oxide (NO)

Relaxes Smooth Muscle

Relaxed muscle disables contraction = dilation

how does no relax smooth muscle
How does NO relax smooth muscle?

NO

Inactivates contraction

Dephosphorylated

Mechanism of smooth muscle relaxation

does hawthorn extract affect no release in endothelial cells
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

nitric oxide production
Nitric Oxide Production

Mechanism of NO production in an endothelial cell

radiotracer assay s mechanism
Radiotracer Assay’s Mechanism

14C-Arginine  14C-Citrulline

  • Measure 14C-Citrulline product
  • Correlate with NO production
does the protocol work
Does the protocol work?

A23197 – calcium ionophore, increases

intracellular [Ca2+] by increasing cell permeability

nitric oxide production1
Nitric Oxide Production

Mechanism of NO production in an endothelial cell

what s next
What’s next?
  • Achieve consistency.
  • How reliable is Dowex resin?
  • Is resin holding the unused C-14-Arginine?
dowex resin
Dowex Resin
  • Ion-exchange
  • Resin overloaded?
my summer methods development1
My Summer…Methods Development
  • Endothelial Cells- Project #1
  • Cardiomyocytes - Project #2, w/Dr. Shannon Long
      • Hypertrophy = thickening of tissue, caused by increased cell size
project 2 cardiomyocytes
Project #2: Cardiomyocytes
  • Heart muscle cells
  • From mice pups
  • Contracts in media
have already found
Have already found …

Epinephrine

Beating cells

have already found1
Have already found…

Epinephrine

NON-Beating cells

hypertrophy
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)

what s next1
What’s Next?
  • Collect more data.
  • If hypertrophy in cells is successful, treat with Hawthorn.
dr filtz s long term goals
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.
acknowledgements
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!