BY: SEAN PAGE

COLLEGE OF HUMANI TI ES AND SOCI AL SCI ENCES

CALI FORNI A STATE UNI VERSI TY, FULLERTON
The Potential Effect of Exercise in
Attenuating the Depression-induced
Deregulation of Cytokines
Interleukin-6 and Interleukin-10
OUTLINE
Introduction
Psychoneuroimmunology (PNI): context and approach
Immunity
Depression
Exercise
Hypothesis
Methods
Behavioral
Biological
Results (expected)
Discussion
Implication
Conclusion



Introduction: PNI
PNI (psychoneuroimmunology): the study of the
interactions between behavior, the brain (or CNS)
and immunity

Goal of the field: understand how various biological
systems work together to effect human health and
disease
Introduction: PNI
Current Perspective/Standards in PNI:
Bidirectional communication:
Brain to Immune Immune to brain

Brain to Immune: stress and associative processing (classic
conditioning). Alter immune function

Immune to Brain: behavioral effects produced by substances
released by the immune system. Alter behavior


Introduction: PNI
Brain (CNS) to Immune Communication:
(1) Peripheral Nervous System Autonomic Nervous System
(ANS)
Introduction: PNI
Brain (CNS) to Immune Communication:
(1) Peripheral Nervous System Autonomic Nervous System
(ANS)

Sympathetic Nervous System (SNS): innervates immune
organs (thymus, bone marrow, spleen, lymph nodes)

SNS nerve terminals release norepinephrine (stress hormone)

Introduction: PNI
Brain (CNS) to Immune Communication:
(2) Endocrine System: collection of glands that secrete
hormones directly into blood circulatory system

Brain can communicate with peripheral organs and immune
compartments releasing hormones and other molecules.
- Glucocorticoids (family of steroid hormones):
stress is often defined by increase levels in blood
- Released by activation of the hypothalamic-
pituitary-adrenal (HPA)-axis
- HPA-axis induced by fight or flight stress
response from the ANS
Immune to Brain Communication:
Both chemical and electrical activity change the in brain as
immune response occurs (esp. hypothalamus)
Cytokines are the primary protein studied- complication-
cytokines are lipophobic (do not cross BBB)
Blood Brain Barrier: cytokines do not usually cross (active
transport or endocytosis purposed)
Administration of Interleukin-1 causes the same behavioral
alterations seen in depression
(reduced activity, novel object exploration, social isolation, food
and water intake, willingness to engage in sexual behavior)


Introduction: PNI
Bidirectional communication:

Introduction: Immunity
Immunity: integrated system of cell communication
regulated by signaling molecules driven by
challenges from foreign substances
Immune response- two types:
(1) innate immunity (older): non-specific resistance to
pathogen (foreign substance)
Primarily physiological, i.e. mucus, and phagocytic, i.e.
macrophages, neutrophils (other phagocytes) engulf pathogen
Two important mechanisms: Inflammation and Acute phase
response
Introduction: Immunity
Immune response- two types:
(2) Adaptive Immunity: specific response to a known
pathogen; acquired defense from antibody
Two distinct processes:
Recognition of non-self foreign substance from B-cell or
macrophage (other antibody generators) and presentation to T-
cells
T-cell mediate immune response through humoral mechanisms
Important to note this is a delayed/slower response relative to
innate (antibody generation and cell proliferation)
Introduction: Immunity
Introduction: Immunity
Inflammation: local response to tissue/immune
challenge characterized by pain, redness, swelling,
temp.

Purpose: to limit damage to tissue; limits pathogen
to specific area for phagocytic destruction

Can be observed 1 to 2 hrs after an immune response
Introduction: Immunity
Acute Phase Response (APR): 8 to 12 hrs. Supports
local defenses and fight infection that has spread

Pro-inflammatory cytokines (immune signaling
proteins): IL-1, IL-6, and Tumor Necrosis Factor- all
trigger APS

APR: increased activity in ANS and levels of
pituitary-adrenal stress hormones increase

Introduction: Depression
Associated with deregulation pituitary-adrenal
system
Patients show elevated levels of glucocorticoids
Pro-inflammatory cytokines produce behavioral
alteration seen in depression (injections in animals;
chemotherapy)
Increase in pro-inflammatory cytokine production
and circulation
Introduction: Depression
Associated with numerous disorders involving
chronic inflammation
Show increased production of pro- vs. anti-
inflammatory cytokines, esp. IL-6
IL-6 concentration/deregulation can be used to
measure the severity of depression
The chronic increase in circulating glucocorticoids
suggested to cause neural injury; has diminishing
benefits that turn to deficits

Introduction: Depression
Potentially mediated in part by pro-inflammatory
cytokines
Patients with Major Depressive Disorder (MDD)
have been shown to have epigenetic changes of
inflammation-genes
The increase in pro- and decrease in anti-
inflammatory cytokines can be reversed by
antidepressants
Introduction: Exercise
Seen as an effective treatment for MDD, as effective
of pharmacological interventions

Benefits: neuroplasticity, neurogenesis,
neuroendocrine regulation, circadian rhythm
regulation, increases in mood, cognitive function
(learning and memory), physiological functions
Introduction: Exercise
Long-term its shown to decrease circulating pro-
inflammatory cytokines
Delays neurodegenerative diseases
Reduces anxiety- and depression-like behaviors
Increases plasma levels of IL-10, an anti-
inflammatory cytokine
Hypothesis
Hypothesis: exercise with counteract the negative
behavioral and immune changes associated with
depression

2 x 2 Design

Depression

No Depression

Exercise

Depression and Exercise group

Exercise Control

No Exercise

Depression Control

General Control

Methods
Sprague-Dawley Albino rats (N=40) with be divided
into groups of 10

Animals are housed 2-per cage (no social isolation)

Day 1-30

Free wheel access
begins

Groups: Exercise and
Depression; Exercise
Control
Day 31

Sucrose Training
session one

Groups: all groups
Day 35

Sucrose Training
session two

Groups: all groups
Day 39

Sucrose Training
session three

Groups: all groups
Day 43

Restraint Stress

Groups: Exercise and
Depression;
Depression control
Day 57

Final day of Restraint
Stress


Day 58

Sucrose Testing

Groups: all groups
Day 60

Blood Collection

Groups: all groups

Methods
Exercise: Voluntary exercise (vs. forced) will be
granted to the exercise condition 30-days before
procedures
7 inch freestanding aluminum running wheels are
placed in the home-cage of exercise conditions
Rats will have continued access to running wheels
for the remainder of the protocol (except for sucrose
testing)
Methods
Sucrose Testing: is an animal model based from
Anhedonia (DSM-V: symptom of depression
characterized by loss of ability to experience
pleasure)
Sucrose testing: requires 3 training and 1 testing
period (which are the same procedure)
Sucrose trainings are administered before the
depression treatment to gather baseline values
Sucrose testing is after the depression treatment for
comparison

Methods
Double-housed animals will be removed from their
home cages and placed into new units
Sucrose presentation: the protocol for sucrose
training- 14 hours of food and water deprivation,
followed by presentation of 1% sucrose solution for a
period of 1 hour
Measurements of sucrose consumption will be
gathered after the 1hr period
Methods
Chronic Mild Stress (CMS) Animal Model of
Depression
CMS: the protocol utilizes restraint stress; 2 hour
period for 14 continuous days
Methods
After CMS-treatment- Sucrose Testing

Sucrose test will gather post-treatment values of
sucrose consumption in all groups for statistical
comparison

Blood Collection with follow the Sucrose testing


Methods
Blood Collection: 23g butterfly needle will be used to
collection .5 ml of blood from the lateral tail vein of
each rat
Methods
Blood Collection samples:
samples will be immediately placed on ice

Centrifuged at 1000g at -4
o
C for 5 minutes

Plasma will be collected and stored in a -80
o
C freezer

IL-6 and IL-10 will then be measured once all samples are
collected
Methods
Enzyme-linked Immunosorbent Assay (ELISA):
direct ELISA will be used
Methods
ELISA: plasma values of IL-6 and IL-10 are outside
the detection range of a standard ELISA and requires
high detection
Results
Depression group: expected to show statistically
significant levels of IL-6 and reduced consumption
of sucrose
Exercise group: expected to show marginal
differences (positive) in IL-6/IL-10 ratio and
marginal increase in sucrose consumption

2 x 2 Design

Depression

No Depression

Exercise

Depression and Exercise group

Exercise Control

No Exercise

Depression Control

General Control

Control Group: expected to show no statistically
significant changes in IL-6 or IL-10 values or sucrose
consumption (may show reductions-sedentary)
Depression and Exercise Group: The levels of IL-6 are
expected to show no change, while IL-10 may increase.
The is expected to be no reduction in sucrose
consumption

2 x 2 Design

Depression

No Depression

Exercise

Depression and Exercise group

Exercise Control

No Exercise

Depression Control

General Control

Discussion
Discussion
Exercise maybe a useful alternative for treatment of
depression
IL-6 production maybe balanced in depressed
patients will mild exercise
There maybe interacting and overlapping pathways
that contribute to both the effects of immunity,
depression and exercise
The extremely high prevalence of inflammatory
disorders co-occuring with depression supports the
expected results (claim)
Discussion
Implications and Conclusion:
Understanding the mechanism of depression is a necessity
Estimates depression as the 4
th
leading cause of disease
burden worldwide and affects about 20% of the population
chronically and more over a life-time
Estimates in economic loss from decreased productivity of the
American population are estimated between 17-44 billion
dollars annually (Center for Disease and Control)
The benefits of integrating preventative strategies in medicine,
especially in a context that is will supported is a necessity