exercise induced endocannabinoid and immune system alterations a thesis submitted to the ...

207 Pages·2010·4.31 MB·English

EXERCISE INDUCED ENDOCANNABINOID AND IMMUNE SYSTEM
ALTERATIONS

A THESIS SUBMITTED TO
THE GRADUATE SCHOOL OF SOCIAL SCIENCES
OF
MIDDLE EAST TECHNICAL UNIVERSITY

BY

RABİA HÜRREM ÖZDURAK

IN PARTIAL FULFILLMENT OF THE REQUIREMENTS
FOR
THE DEGREE OF DOCTOR OF PHILOSOPHY
IN
PHYSICAL EDUCATION AND SPORTS

NOVEMBER 2009
Approval of the Graduate School of Social Sciences

Prof. Dr. Sencer Ayata
Director

I certify that this thesis satisfies all the requirements as a thesis for the degree of
Doctor of Philosophy.

Assoc. Prof.Dr. Settar Koçak
Head of Department

This is to certify that we have read this thesis and that in our opinion it is fully
adequate, in scope and quality, as a thesis for the degree of Doctor of
Philosophy.

Prof. Dr. Feza Korkusuz
Supervisor

Examining Committee Members

Prof. Dr. Sevim Ercan (GAZI, MP)
Prof. Dr. Feza Korkusuz (METU, PES)
Prof. Dr. Duygu Uçkan (HU,BMT)
Assoc. Prof. Dr. Petek Korkusuz (HU,HE)
Assist. Prof. Dr. Sadettin Kirazcı (METU, PES)
I hereby declare that all information in this document has been obtained
and presented in accordance with academic rules and ethical conduct. I also
declare that, as required by these rules and conduct, I have fully cited and
referenced all material and results that are not original to this work.

Name, Surname: Rabia Hürrem Özdurak
Date:
Signature:

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ABSTRACT

EXERCISE INDUCED ENDOCANNABINOID AND IMMUNE SYSTEM
ALTERATIONS

Özdurak, Rabia Hürrem
Ph.D., Department of Physical Education and Sports
Supervisor : Prof. Dr. Feza Korkusuz
November 2009, 186 pages

Endocannabinoid and immune system alterations at moderate (18 m/min)
and endurance (32 m/min) exercise intensities were assessed and compared to
controls. Rats were exercised for 60 minutes/day, 5 days/week for 16 weeks.
Immune effector cell proportions (T cell subtypes, B cells, NK cells, and
neutrophils) and endocannabinoid serum levels were determined. Anandamide
(ANA) and 2 arachidonyl-glycerol (2-AG) serum levels increased with
endurance type of exercise. mRNA expression of the CB1 receptor increased
together with ANA in the same group. Apoptotic index increased while immune
effector cells responded divergently. B lymphocyte percentage decreased while
T lymphocyte and NK cell percentage increased in blood. CD8+ subtypes
increased whereas CD11b+ cell and CD25+ cell numbers decreased in the spleen
in the endurance type of exercise group.
Rats were grouped as the control, the endurance type of exercise, the
AM281 (CB1 receptor antagonist) and the AM281+AM630 (CB2 receptor
antagonist) groups in the second part of the study. Flow cytometry and
microarray analyses of the spleen and the thymus were conducted. Endurance
type of exercise associated significantly to immunological changes particularly
to that of the T lymphocytes. T lymphocytes increased whereas cytolytic T
lymphocytes decreased in blood. T cell and double positive T cell percentages
significantly increased in the spleen. Activated T cells and NK like T cells
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furthermore decreased in the spleen. AM281 and/or AM630 could partially
reverse the effect of exercise in blood but not in the spleen. Alterations in the
thymus were not observed. Exercise altered 302 genes, some of them related
with the immune system. Up-regulation of heat-shock protein coding genes was
the most significant ones.

Keywords: Anandamide, 2-arachidonylglycerol, T lymphocytes, exercise
intensity, spleen
v
ÖZ

FĠZĠKSEL EGZERSĠZE DAYALI ENDOKANNABĠNOĠD VE
ĠMMÜN SĠSTEM DEĞĠġĠKLĠKLERĠ

Rabia Hürrem Özdurak
Doktora, Beden Eğitimi ve Spor Bölümü
Tez Yöneticisi: Prof. Dr. Feza Korkusuz
Kasım 2009, 186 sayfa

Endokannabinoid ve imün sistemdeki değiĢiklikler orta Ģiddet (18 m/min)
ve endurans (32 m/min) egzersizde ölçülerek control grubu ile
karĢılaĢtırılmıĢtır. Sıçanlar günde 60 dakika ve haftada 5 gün olacak Ģekilde 16
hafta boyunca egzersiz yaptırılmıĢlardır. Imün efektör hücre oranları (T hücre
alt tipleri, B hücreler, NK hücreler ve nötrofiller) ve endokannabinoid serum
düzeyleri belirlenmiĢtir. Anandamid (ANA) ve 2 araĢidonilgliserol (2-AG)
serum düzeyleri endurans tipi egzersiz ile artmıĢtır. CB1 reseptörünün mRNA
ekspresyonu ANA ile aynı grupta artmıĢtır. Kanda imün efektör hücreler
değiĢken tepkiler verirken apoptotik indeks artmıĢtır. B lenfosit yüzdesi
azalırken T lenfosit ve NK hücre yüzdesi artmıĢtır. Endurans tipi egzersiz
sonrasında CD8+ alt tipi artarken CD11b+ hücre ve CD25+ hücre sayıları
dalakta azalmıĢtır.
Sıçanlar çalıĢmanın ikinci bölümünde control, endurans tipi egzersiz,
AM281 (CB1 reseptör antagonisti) ve AM281+AM630 (CB1 ve CB2 reseptör
antagonisti) olacak Ģekilde gruplandırılmıĢlardır. Dalak ve timus dokularında
akıĢkan hücre tekniği ve microarray analizleri yapılmıĢtır. Endurans tipi
egzersiz T hücreleri baĢta olmak üzere imünolojik değiĢiklikler ile
iliĢkilendirilmiĢtir. Kanda T lenfositler artarken sitolitik T lenfositler azalmıĢtır.
Double pozitif T hücreler dalakta anlamlı Ģekilde artmıĢtır. Dalakta aktive T
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hücreler ile NK benzeri T hücrelerde azalmıĢtır. AM281 ve/veya AM630 kanda
egzersizin etkisini kısmen ortadan kaldırabilirken dalakta etkili olamamıĢtır.
Timusta değiĢiklik gözlenmemiĢtir. Egzersiz 302 genin ekspresyonunda
değiĢikliğe sebep olmustur. Ġçerisinde bağıĢıklık sistemi ile iliĢkili olan
genlerinde bulunduğu grupta en çok değiĢikliği heat shock proteinlerini
kodlayan genler oluĢturmuĢtur.

Keywords: Anandamide, 2-arachidonylglycerol, exercise intensity, T
lymphocytes, spleen

vii
To My Dad

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ACKNOWLEDGEMENT

I would like to express my deepest gratitude and thanks to my supervisor
Prof. Dr. Feza Korkusuz for his invaluable supervision, patience, insight,
suggestions, comments and red pen not only during this thesis, but also in my
life. The completion of this degree would have been impossible without his
guidance.

I would like to express my very special thanks to Assoc. Prof. Dr. Petek
Korkusuz for inspiring me and opening a very interesting window in to my
science life. and for her endless patient and assistance during this thesis.

I also wish to thank the members of examining committee, Prof. Dr.
Sevim Ercan, Prof. Dr. Duygu Uçkan and Assist. Prof.Sadettin Kirazcı for their
valuable suggestions and infinitive support throughout this study.I also wish to
thank Assoc. Prof. Dr.Çetin for his valuable support and assistance during this
study.

I would like to express my special thank to Dr. Dilara Zeybek and Irem
Akar for their support and valuable comments during this thesis.

I wish to extend thanks to the staff of the Department of Medical Biology
of Hacettepe University, Heamatology Laboratory of Gazi University, Pediatric
Heamatology Laboratory and Animal Research and Production Laboratory of
Hacettepe University, Medical Center and Molecular Biology and
Biotechnology R & D Center of Middle East Technical University for
allowance to use their facility, machinery and resources. In addition, I would
like to express my special thanks to Dr. Tamay ġeker, Emine Kılıç, Uğur
Akpulat and Sezin Dağdeviren for their valuable comments, suggestions and
sincerity nearby their sleepless nights with enourmous patience.
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Special thanks are to my husband Mehmet, my friends Sevil Ünlütürk and
Dilek AlataĢ for their moral support and infinite patience in completing this
thesis and taking care to my little angel Defne while I was busy with this thesis.

Finally, I would express my special thanks to Middle East Technical
University Scientific Research (Grant #2007(R)081103), the Scientific and
Technical and Research Council of Turkey (Grant # 107S211) and Hacettepe
University Scientific Research (Grant # 07-01-101-010) for their grants and
support

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