the timing of fluoxetine, simvastatin and ascorbic acid administration in a post-ischemic stroke ...

101 Pages·2016·1.01 MB·English

by  Neal Verma

THE TIMING OF FLUOXETINE, SIMVASTATIN AND ASCORBIC ACID
ADMINISTRATION IN A POST-ISCHEMIC STROKE ENVIRONMENT AFFECTS
INFARCT VOLUME AND HEMORRHAGIC TRANSFORMATION FREQUENCY

A thesis submitted in partial fulfillment of the
requirements for the degree of
Master of Science

By

NEAL RAJ VERMA
Bachelor’s of Science, University of Dayton, 2009
Medical Degree, Wright State University, 2015

2016
Wright State University
WRIGHT STATE UNIVERSITY

GRADUATE SCHOOL

DATE OF DEFENSE
MAY 18, 2016

I HEREBY RECOMMEND THAT THE THESIS PERPARED UNDER MY SUPERVISION
BY Neal Raj Verma ENTITLED The Timing Of Fluoxetine, Simvastatin and Ascorbic
Acid Administration in a Post-Ischemic Stroke Environment Affects Infarct Volume
and Hemorrhagic Transformation Frequency BE ACCEPTED IN PARTIAL
FULFILLMENT FOR THE DEGREE OF Master of Science.

Adrian Corbett, Ph.D.
Thesis Director

Christopher N. Wyatt, Ph.D.
Interim Chair
Department of Neuroscience, Cell Biology and Physiology

Committee on
Final Examination

Adrian Corbett, Ph.D.

Debra Mayes, Ph.D.

Mary White, Ph.D.

Robert E. W. Fyffe, Ph.D.
Vice President for Research and
Dean of the Graduate School
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ABSTRACT

Verma, Neal Raj. M.S., Department of Neuroscience, Cell Biology and Physiology,
Wright State University, 2016. The Timing of Fluoxetine, Simvastatin and Ascorbic
Acid Administration in a Post-Ischemic Stroke Environment Affects Infarct Volume
and Hemorrhagic Transformation Frequency.

Previous animal experiments have indicated that administration of
fluoxetine and simvastatin at 20-26 hours post-stroke decreases the volume of
ischemic infarcts. This experiment expanded on previous experiments by adding
ascorbic acid to the post-stroke regimen, initiating simvastatin pre-stroke, and
adding a third initiation time frame (48-54 hours).
Male retired breeder Sprague-Dawley rats were on simvastatin for 7 days
prior to stroke induction. Combined medications of 5 milligrams/kilogram of
fluoxetine, 1 milligram/kilogram of simvastatin and 20 milligrams/kilogram of
ascorbic acid were orally administered at 6-12 hours, 20-26 hours, or 48-54 hours,
respectively, following stroke induction.
Adult rats that were treated 20-26 hours post-stroke showed a decrease in
infarct volume (15.67 ± 5.622 millimeters cubed, P=0.0098) compared to the
control. The combination of simvastatin, fluoxetine and ascorbic acid decreased the
relative risk (RR=0.3704 (95% confidence interval 0.0987 to 1.3905, p-value =
0.1411) of bleeding after ischemic stroke if initiated 20-26 hours after stroke
induction in rats.
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TABLE OF CONTENTS

Page

I. INTRODUCTION……………………………………………………………………………………………1
Stroke…...........................................................................................................................1
Stroke Research………………………………………………………………………………...….9
Pharmacologic Treatment…………………………………………………………………..13
Post Ischemic Infarct Bleeding……………………………………………………………25
Hypothesis………………………………………………………………………………………….28
Specific Aims……………………………………………………………………………………….29
II. MATERIALS AND METHODS………………………………………………………………………30
Stroke Induction…………………………………………………………………………………30
Post-stroke Treatment………………………………………………………………………..33
Infarct Analysis…………………………………………………………………………………..33
III. RESULTS…………………………………………………………………………………………………..37
Experiment Results…………………………………………………………………………….37
IV. DISCUSSION………………………………………………………………………………………………68
Results Summary………………………………………………………………………………..68
Future Experiments……………………………………………………………………………79
Conclusions…………………………………………………………………………………………81
REFERENCES………………………………………………………………………………………………….84

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LIST OF FIGURES

Figure Page

1. Figure 1……………………………………………………………………………………………………60
2. Figure 2……………………………………………………………………………………………………62

v
LIST OF TABLES

Table Page

1. Table 1………………………………………………………………………………………………………39
2. Table 2………………………………………………………………………………………………………48
3. Table 3………………………………………………………………………………………………………52
4. Table 4………………………………………………………………………………………………………58
5. Table 5………………………………………………………………………………………………………65

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LIST OF GRAPHS

Graph Page

1. Graph 1…………………………………………………………………………………………………….50
2. Graph 2…………………………………………………………………………………………………….54

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ACKNOWLEDGEMENTS

There are many individuals who I would like to acknowledge for making this
project possible. First, Dr. Adrian Corbett has been most generous in allowing me to
work in her laboratory and to undertake this project. Her steady guidance has been
integral in seeing this project through, and I cannot speak enough about her
willingness to help in any way possible. It is also worth noting that she is a
wonderfully vivacious human being and a pleasure to be around. I would also like to
extend this acknowledgement to my fellow students working in the laboratory as
well. The friendliness and congeniality that I received on behalf of my fellow lab
students makes coming to work all the more worthwhile.
I would also like to acknowledge Dr. Larry Ream for his guidance throughout
my academic career. Dr. Ream’s Anatomical Sciences program has been a windfall
for me at several times during my academic career, and I would like to extend my
acknowledgment to the entire department of Neuroscience, Cell Biology and
Physiology.
My final acknowledgement goes to my mother, father and younger brother.
Without their support, encouragement and guidance, none of my accomplishments
would have come to pass. I owe them everything, and I can only hope to live up to
the example that they have set before me.

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For my loving mother, father and brother

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I. INTRODUCTION
STROKES
In medical terminology, a stroke is defined as a loss of blood flow to brain
tissue (National Stroke Association (NSA), 2016). Blood carries vital nutrients,
including oxygen, to brain tissue. When brain tissue is denied blood, it dies, and
the abilities that were under control of that particular area of the brain are lost.
The functions of the brain are localized to various areas within the brain, and the
abilities that are lost after a stroke typically correlate to the location of the
stroke and the amount of brain tissue that dies (NSA 2016).
Statistically, strokes are a devastating affliction on the American
population. Every year, 800,000 people will experience a stroke (new or
recurrent). Strokes are the fifth leading cause of death in the United States, and
they are the leading cause of disability (NSA 2016). Broken down by gender,
40% of stroke deaths are in males and 60% are in females (American Heart
Association (AHA)/American Stroke Association (ASA) 2016). There are many
risk factors that predispose individuals to strokes. They include being
overweight, a lack of physical activity, consuming large quantities of alcohol and
the use of cocaine or methamphetamines (Mayo Clinic 2016). Because strokes
are frequently permanently disabling, there is a tremendous need for research
into post‐stroke care. Specifically, the ability to rapidly accelerate neuro‐
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