Depression and Anxiety

Renae Geier and Leila Shinn

NUTR 561
February 24th, 2015

Outline
Introduction

Gut-Brain Axis: How the Microbiome Influences Anxiety and Depression

Research Articles

Ingestion of Lactobacillus strain regulates emotional behavior and

central GABA receptor expression in a mouse via the vagus nerve
Assessment of psychotropic-like properties of a probiotic formulation
(Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 in
rats and human subjects)

Strengths and Limitations

Future Directions

Outline
Introduction

Gut-Brain Axis: How the Microbiome Influences Anxiety and Depression

Research Articles

Ingestion of Lactobacillus strain regulates emotional behavior and

central GABA receptor expression in a mouse via the vagus nerve
Assessment of psychotropic-like properties of a probiotic formulation
(Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 in
rats and human subjects)

Strengths and Limitations

Future Directions

Wang et al., 2014

The Hypothalamic-Pituitary-Adrenal (HPA) Axis

Stephens et al., 2012

Stress and the Microbiota

Alterations in HPA axis function
Depressive episodes are associated with
dysregulation of the HPA axis
Gut microbiota plays a role in the programming
of the HPA axis in early life and stress reactivity
over the lifespan
Inflammation and intestinal permeability
Direct influences on stress circuits
Activation of stress circuits through vagal
pathways
Enteric nervous system

Gut-Brain Axis and Behavior

Foster et al., 2013

Gut-Brain Axis and Behavior

GF housing and antibiotic treatment reduced
anxiety-like behavior

Foster et al., 2013

Gut-Brain Axis and Behavior

Probiotics influence anxiety-like and depressivelike behaviors
Infection and gut inflammation increase anxietylike behavior

Gut-Brain Axis and Behavior

Inflammatory state influences behavior

Foster et al., 2013

Clinical Evidence of Probiotic Use for Mood

and Anxiety Symptoms to Date
Little published work on effects of probiotics on
depression or anxiety symptoms in humans
Evidence that probiotics have similar
antidepressive and anxiolytic effects as those
observed in preclinical studies

Messaoudi, M. et al. (2011) Beneficial psychological effects of a

probiotic formulation (Lactobacillus helveticus R0052 and
Bifidobacterium longum R0175) in healthy human volunteers. Gut
Microbes 2, 256-261
Benton, D. et al. (2007) Impact of consuming a milk drink
containing a probiotic on mood and cognition. Eur .J. Clin. Nutr. 61,
355-361

Chronic fatigue syndrome (CFS)

Gut-brain Axis and Neurochemistry

Do changes in behavior mediated by microbiota
result in long-term changes in central signaling
systems?
GABAergic signaling

Papers to be Presented
Review: Gut-brain axis: how the microbiome influences
anxiety and depression
Preclinical study: Ingestion of Lactobacillus strain
regulates emotional behavior and central GABA receptor
expression in a mouse via the vagus nerve
Preclinical and Clinical study: Assessment of
psychotropic-like properties of a probiotic formulation
(Lactobacillus helveticus R0052 and Bifidobacterium
longum R0175) in rats and human subjects

Outline
Introduction

Gut-Brain Axis: How the Microbiome Influences Anxiety and Depression

Research Articles

Ingestion of Lactobacillus strain regulates emotional behavior and

central GABA receptor expression in a mouse via the vagus nerve
Assessment of psychotropic-like properties of a probiotic formulation
(Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 in
rats and human subjects)

Strengths and Limitations

Future Directions

Ingestion of Lactobacillus strain

regulates emotional behavior and
central GABA receptor expression in
a mouse via the vagus nerve
Bravo et al., 2011

Introduction
Problem: There is a lack of direct evidence that
commensal GI microbes can affect anxiety and
depression

Introduction
Problem: There is a lack of direct evidence that
commensal GI microbes can affect anxiety and
depression

Could a lactic acid bacterium have an effect on

anxiety and depression in healthy animals?

Introduction
Problem: There is a lack of direct evidence that
commensal GI microbes can affect anxiety and
depression

Could a lactic acid bacterium have an effect on

anxiety and depression in healthy animals?
If so, how does this bacterium exert its
effects?

Experimental Approach
Tests:

Behavior:

Mice: BALB/c,
adult, male

SIH: stress-induced
hyperthermia

EPM: elevated plus

maze

FST: forced swim test

Vagotomized
mice

Open field test

Fear conditioning

Hormones:

Corticosterone levels

mRNA
expression:

GABAA

GABAB

Treatment: Lactobacillus rhamnosus JB-1

Experimental Approach
Tests:

Behavior:

Mice: BALB/c,
adult, male

SIH: stress-induced
hypothermia

EPM: elevated plus

maze

FST: forced swim test

Vagotomized
mice

Open field test

Fear conditioning

Hormones:

Corticosterone levels

mRNA
expression:

GABAA

GABAB

Treatment: Lactobacillus rhamnosus JB-1

Experimental Approach
Tests:

Behavior:

Mice: BALB/c,
adult, male

SIH: stress-induced
hyperthermia

EPM: elevated plus

maze

FST: forced swim test

Vagotomized
mice

Open field test

Fear conditioning

Hormones:

Corticosterone levels

mRNA
expression:

GABAA

GABAB

Treatment: Lactobacillus rhamnosus JB-1

Experimental Approach
Tests:

Behavior:

Mice: BALB/c,
adult, male

SIH: stress-induced
hyperthermia

EPM: elevated plus

maze

FST: forced swim test

Vagotomized
mice

Open field test

Fear conditioning

Hormones:

Corticosterone levels

mRNA
expression:

GABAA

GABAB

Experimental Approach
Tests:

Behavior:

Mice: BALB/c,
adult, male

SIH: stress-induced
hyperthermia

EPM: elevated plus

maze

FST: forced swim test

Vagotomized
mice

Open field test

Fear conditioning

Hormones:

Corticosterone levels

mRNA
expression:

GABAA

GABAB

Treatment: Lactobacillus rhamnosus JB-1

GABAA receptor

ClRichter et al., 2012

GABAB receptor

Bettler et al., 2004

Experimental Approach
Tests:

Behavior:

Mice: BALB/c,
adult, male

SIH: stress-induced
hyperthermia

EPM: elevated plus

maze

FST: forced swim test

Vagotomized
mice

Open field test

Fear conditioning

Hormones:

Corticosterone levels

mRNA
expression:

GABAA

GABAB

Treatment: Lactobacillus rhamnosus JB-1

Behavioral activity

Corticosterone levels

GABAB1b mRNA
cingulate
cortex

pre-frontal
cortex

amygdala

locus
coeruleus

hippocampus

GABAB1b mRNA
cingulate
cortex

pre-frontal
cortex

amygdala

locus
coeruleus

hippocampal

GABAA2 mRNA

Behavioral activity of vagotomized mice

GABAA mRNA expression in hippocampus

GABAA2 mRNA in vagotomized mice

GABAA1 mRNA in vagotomized mice

Discussion
L. rhamnosus JB-1 can have a direct effect on behavior
and physiology related to anxiety and depression in
healthy animals and is dependent on the vagus nerve
Tests:

Behavior:

Mice: BALB/c, adult,

male

SIH: stress-induced hyperthermia

EPM: elevated plus maze

FST: forced swim test

Vagotomized mice

Open field test

Fear conditioning

Hormones:

Corticosterone levels

mRNA
expression:

GABAA

GABAB

?
?

Outline
Introduction

Gut-Brain Axis: How the Microbiome Influences Anxiety and Depression

Research Articles

Ingestion of Lactobacillus strain regulates emotional behavior and

central GABA receptor expression in a mouse via the vagus nerve
Assessment of psychotropic-like properties of a probiotic formulation
(Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 in
rats and human subjects)

Strengths and Limitations

Future Directions

Assessment of psychotropic-like
properties of a probiotic formulation
(Lactobacillus helveticus R0052 and
Bifidobacterium longum R0175) in
rats and human subjects
Messaoudi et al., 2011

Introduction
Problem: There is a lack of evidence that oral
intake of probiotics may have beneficial consequences
on mood and psychological distress

Introduction
Problem: There is a lack of evidence that oral
intake of probiotics may have beneficial consequences
on mood and psychological distress
Objective: To investigate the anxiolytic-like activity
of a probiotic formulation (PF) in rats and its possible
effects on anxiety, depression, stress, and coping
strategies in healthy humans

Experimental Approach: Preclinical Trial

36 male Wistar rats weighing 200 g

Three groups (n=12):

Probiotic preparation (PF)
Placebo (0.5% methylcellulose solution)
Diazepam (Valium 1%)
Free access to food pellets and tap water (until day
before anxiety testing)
Conditioned defensive burying test

Products
Mixture of freeze-dried lactic acid bacteria and

excipients
Bacterial Strains

Excipients

L. helviticus R0052
B. longum R0175

Xylitol
Maltodextrin
Flavor
Malic Acid

All products freshly prepared daily and

administered by gavage (5 ml/kg)

Results
Statistical analysis showed a group difference in the

stress/anxiety score
Lower in rats treated with PF and diazepam than
with vehicle

Experimental Approach: Clinical Trial

55 Caucasian men and women
Double-blind, controlled, randomized, parallel
study lasting 30 days
Preliminary examination, baseline (14 d), and
follow-up (30 d after baseline)
Two groups:
Test product (PF) or placebo (PL)

Testing Methods

Hopkins Symptom Checklist-90 (HSCL-90)

Hospital Anxiety and Depression Scale (HADS)
Perceived Stress Scale (PSS)
Coping Checklist (CCL)
Urinary free cortisol

Results

Global severity index percentage change was higher

in the PF compared to the PL (p<0.05)

Improved somatization, depression, and angerhostility subscales

Results

Percentage changes in HADS and HADS-A were

higher in the PF treated subjects

The HADS-D score for the PF decreased
significantly

Results

PL subjects increased their positive re-evaluation

score between BL and FU

PF subjects decreased their self-blame score and
displayed higher problem solving score between the
two test sessions

Summary and Conclusions

Consumption of the PF containing L. helveticus

R0052 and B. longum R0175 in combination

mitigated psychological distress in three tests
without displaying any adverse effects.
Gut microbiota play a role in stress. anxiety, and
depression (via the ENS and CNS)
Probiotics may offer a useful therapeutic approach to
neuropathological disorders and/or as adjunct
therapies in psychiatric disorders

Outline
Introduction

Gut-Brain Axis: How the Microbiome Influences Anxiety and Depression

Research Articles

Ingestion of Lactobacillus strain regulates emotional behavior and

central GABA receptor expression in a mouse via the vagus nerve
Assessment of psychotropic-like properties of a probiotic formulation
(Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 in
rats and human subjects)

Strengths and Limitations

Future Directions

Strengths

Limitations

molecular mechanisms are

being developed
multiple methods of analysis
and testing in both studies
links between preclinical
and clinical

limited ethnic and age

diversity in clinical study
only used male mice in the
preclinical studies
sexual differences
mouse studies are difficult to
translate to humans

Outline
Introduction

Gut-Brain Axis: How the Microbiome Influences Anxiety and Depression

Research Articles

Ingestion of Lactobacillus strain regulates emotional behavior and

central GABA receptor expression in a mouse via the vagus nerve
Assessment of psychotropic-like properties of a probiotic formulation
(Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 in
rats and human subjects)

Strengths and Limitations

Future Directions

Future Studies

More clinical trials

Gender and ethnicity
Combination with other treatments
Other probiotic strains
Molecular mechanisms
How exactly are these probiotics exerting their
effects? Must they be active cultures?