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The Short-Term Effect of

Ovariectomy on Vascular Reactivity in Mouse


Mesenteric Arteries

James Daniel Fleming


Faculty Mentor:
Brent J. F. Hill Ph.D.
Committee Members:
Jeff Padberg Ph.D.
Ben Rowley Ph.D.

General Outline

Background
Methods
Results
Conclusion
Acknowledgements

Background

General Outline

Cardiovascular Disease and Hypertension


Voltage-gated calcium channels (CaL)

Methods
Results
Conclusion
Acknowledgements

Cardiovascular Disease (CVD) in the


Population
Leading cause of mortality in men and women
Male mortality has been steadily decreasing,
but not women
Hypertension is a leading cause of CVD
(stroke, heart attack, coronary artery disease,
venous thromboembolism)
72 million Americans with hypertension
Well controlled in 25 million

Factors Contributing to Hypertension


(Coylewright et al., 2008)

BP Differences Between Pre and


Postmenopausal Women
(Staessen et al., 1989)

Estrogen and Testosterone Contributions to Hypertension


(Dubey et al., 2002)

General Outline
Background
Cardiovascular Disease and Hypertension
Voltage-gated calcium channels (CaL)

Methods
Results
Conclusion
Acknowledgements

Smooth Muscle Contraction


VGC
C

Schematic of Voltage-Gated Ca 2+ Channel


(Lacinova, 2005)

Questions
Does OVX alone decrease vascular reactivity
(increased tonicity) in resistance arteries?
Studies using OVX induce pathology
(hypertension)

What effect does E2 have on mouse


morphology?
Body
Uterus
Heart

Other Questions
Part of large project with many others
If OVX mesentery demonstrates increased
voltage-gated Ca2+ channel expression (Rosss
project)
If OVX mesentery demonstrates increased Ca2+
influx (Charmains project)

Significance
41% of postmenopausal have hypertension
75% of women over 60 are hypertensive

Only ~20-45% of hypertensive postmenopausal


women have fully normalized blood pressure
Hypertension = upregulation of Ca L channels
Use of 3 subunit to decrease CaL expression
Find a new route of intervention for the long
term treatment of hypertension

Purpose
To study E2s effects on mouse morphology
and physiology
To determine E2s effects on vascular
reactivity in resistance arteries

General Outline
Background
Methods
Mouse dissection
Western blots

Results
Conclusion
Acknowledgements

Methods

Mice underwent OVX or SHAM surgery at 8 weeks


Euthanized at 8, 10, 12, or 14 weeks
Blood plasma collected for E2 levels
Weights measured
Uterus, Heart, and Body

Dissected mesentery and aorta


Right tibia lengths measured
Hind limbs saved for future study
Bone studies (osteoporosis)

Sample size limitations

Heart Dissection
Hearts dissected for
weight measurement

Whole Heart
RV and LV combined
Right Ventricle
Left Ventricle

Aorta dissection

Mesentery Dissection

Mesentery Dissection

Mesentery Dissection

Mesentery Dissection

3
1

3
4
1

Methods-Vascular Reactivity
Mesenteric arteries
Second order
150-200 m lumen diameter

Cannulated and perfused


with PSS
Superfused with PSS,
FPL64176
Measured change in lumen
diameter
One-way ANOVA

Background
Methods
Results

General Outline

Morphological Changes
Vascular Reactivity

Conclusion
Acknowledgements

Change in Body and Uterine Masses

OVX mice have smaller uteri than


SHAM mice. Uterine mass in mice
that were zero, two, four, or six weeks
post SHAM or OVX surgery. OVX mice
demonstrated significantly smaller
uterine masses than SHAM mice (oneway ANOVA; p<0.0001)(n=4; n=8 for
2-week SHAM). * indicates p<0.05.

Age effect on body masses


between OVX and SHAM mice.
There was an age effect observed in
both OVX (one-way ANOVA;
p=0.0013) and SHAM groups (oneway ANOVA; p=0.0022). However,
there were no significant differences
between age-matched groups at two
weeks (t-test; p=0.0965), four weeks
(t-test; p=0.8244), or six weeks (ttest; p=0.2356) post surgery (n=4;
n=8 for 2-week SHAM).
*
*
*

Mouse Heart Weights


Ovariectomy has no significant
effect on whole-heart masses.
Whole-heart masses in mice that
were zero, two, four, or six weeks
post SHAM or OVX surgery. While
there is a trend for age effect, there
was no significant difference
between OVX and SHAM mice at any
time point (one-way ANOVA;
p=0.4785)(n=4; n=8 for 2-week
SHAM).
Ovariectomy has no significant
effect on combined ventricular
masses. Combined ventricular
masses in mice that were zero, two,
four, or six weeks post SHAM or OVX
surgery. There was no significant
difference between OVX and SHAM
mice at any time point (one-way
ANOVA; p=0.7731)(n=4; n=8 for 2-

Mouse Heart Weights


Ovariectomy has no significant
effect on right ventricular
masses. Right ventricular masses
in mice that were zero, two, four, or
six weeks post SHAM or OVX surgery.
There was no significant difference
between OVX and SHAM mice at any
time point, though OVX right
ventricular masses trended heavier
(one-way ANOVA; p=0.1820)(n=4;
n=8 for 2-week SHAM).
Ovariectomy has no significant
effect on left ventricular masses.
Left ventricular masses in mice that
were zero, two, four, or six weeks
post SHAM or OVX surgery. There
was no significant difference between
OVX and SHAM mice at any time
point (one-way ANOVA; p=0.6495)
(n=4; n=8 for 2-week SHAM).

Vascular Reactivity

n=2
n=3
n=4
n=3
n=2
n=1

Ovariectomy has no significant effect on contractility in


second order mesenteric arteries. Percent change in lumen
diameter in mice that were zero, two, four, or six weeks post SHAM
or OVX surgery. There was no significant difference between OVX
and SHAM mice at any time point (one-way ANOVA; p=0.3912),
though there is a trend for age effect. n-values are indicated on the

Vascular Reactivity

n=2
n=3
n=4
n=3
n=2
n=1

Ovariectomy has no significant effect on contractility in second


order mesenteric arteries. Percent change in lumen diameter in mice
that were zero, two, four, or six weeks post SHAM or OVX surgery. There
was no significant difference between OVX and SHAM mice at any time
point (one-way ANOVA; p=0.1611), though there is a trend for age effect.
n-values are indicated on the figure.

General Outline

Background
Methods
Results
Conclusion
Acknowledgements

Conclusions
Age effect on body mass, but no
treatment effect
Treatment effect on uterine mass
Trends, but no significant effect, on
heart masses

Conclusions
No significant effect on vascular
reactivity
Limited sample size
Conduit v. resistance arteries
Generation of spontaneous tone at 80
mm Hg
Tone increased 25% from 60 mm Hg

General Outline

Background
Methods
Results
Conclusion
Acknowledgements

Acknowledgements
My committee members:
Brent J.F. Ph.D.
Jeff Padberg Ph.D.
Ben Rowley Ph.D.

Other Faculty:
Kristie Hicks Ph.D.
Mike Martin M.D.

Lab Members:

Renee Jordan
Ross Gray
Charmain Fernando
Edouard Niyonsaba
Mohamed Idrissa Moussa

UCA Biology Department


National Institutes of Health
Arkansas INBRE

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