Effects of Hawthorn Crataegus Plant Extracts on Endothelial Cells and Cardiomyocytes

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Effects of Hawthorn Crataegus Plant Extracts on
Endothelial Cells and Cardiomyocytes

• By Linda Phan
• Mentor Dr. Theresa Filtz

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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

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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?
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Congestive Heart Failure

• Causes
• Hypertension high blood pressure
• Hypertrophy thickening of tissue, caused by
  increased cell size

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My Summer Methods Development

• Endothelial Cells- Project 1
• Hypertension high blood pressure
• Cardiomyocytes - Project 2.
• w/Dr. Shannon Long

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Project 1 Endothelial Cells
Endothelial Cells
Nitric Oxide (NO)
Relaxes Smooth Muscle
Relaxed muscle disables contraction dilation
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How does NO relax smooth muscle?
NO
?
Inactivates contraction
Dephosphorylated
Mechanism of smooth muscle relaxation
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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 Hawthorns effects

EA.hy926 cells in culture From the Human
Umbilical Vein
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Nitric Oxide Production
Mechanism of NO production in an endothelial cell
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Radiotracer Assays Mechanism
14C-Arginine ? 14C-Citrulline

• Measure 14C-Citrulline product
• Correlate with NO production

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Does the protocol work?
A23197 calcium ionophore, increases
intracellular Ca2 by increasing cell
permeability
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Nitric Oxide Production
Mechanism of NO production in an endothelial cell
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Edited diagram by Cayman Chemical
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Results
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Results
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Whats next?

• Achieve consistency.
• How reliable is Dowex resin?
• Is resin holding the unused C-14-Arginine?

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Dowex Resin

• Ion-exchange
• Resin overloaded?

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Thin Layer ChromatographyComing Soon!
Before Dowex
After Dowex
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My Summer Methods Development

• Endothelial Cells- Project 1
• Cardiomyocytes - Project 2, w/Dr.
  Shannon Long
• Hypertrophy thickening of tissue, caused by
  increased cell size

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Project 2 Cardiomyocytes

• Heart muscle cells
• From mice pups
• Contracts in media

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Contracted vs. Relaxed

• Contracted

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Contracted vs. Relaxed

• Relaxed

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Contracted vs. Relaxed

• Relaxed

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Contracted vs. Relaxed

• Contracted

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Have already found
Epinephrine
Beating cells
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Have already found
Epinephrine
NON-Beating cells
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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)
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Does Phenylephrine induce hypertrophy?

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Results
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Whats Next?

• Collect more data.
• If hypertrophy in cells is successful, treat with
  Hawthorn.

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Dr. Filtzs 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.

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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!