THE EFFECT OF EXTRA VIRGIN OLIVE OIL ON IL6 AND CRP IN HIGH FAT DIET RATS
Thesis
Submitted as partial fulfilling of the requirement for Master Degree of Biomedical Science
SALEM. M. ALJATLAWI 22010112419054
FACULTY OF MEDICINE DIPONEGORO UNIVERSITY
SEMARANG
2014
APPROVAL PAGE
THE EFFECT OF EXTRA VIRGIN OLIVE OIL ON IL6 AND CRP IN HIGH FAT DIET RATS
Arranged by
SALEM. M. ALJATLAWI 22010112419054
Semarang jun 2014 Approved by:
Supervisor I Supervisor II
Prof. Dr. Lisyani BSuromo. Sp.Pk(K) Dr.dr.RAKisdjamiatun. RMD. MSC
NIP. 130354869 NIP. 196401301990032001
Approved by,
Head of Master Degree Program in Biomedical Science Faculty of Medicine Diponegoro University
Prof. Dr. dr. Tri Nur Kristina. DMM. M.Kes
iii
DECLARATION
I herebydeclarethatthisthesisis my own workand has not been submitted in any form for another degree or diploma at any university or other institution of tertiary education, there are noelementsbelongingPlagiarismforth inDecree No. 17 of 2010. Information derived from the published or unpublished work of others has been acknowledged in the text and a list of reference is given.
Semarang, 30thMay 2014
SALEM. M. ALJATLAWI
v
CURRICULUM VITAE
A. IDENTITY
Name : SALEM. M. ALJATLAWI
Sex : Male
Date & place of birth: Zliten , Libya, jun 14th, 1987 Religion : Moslem
Address : Zliten , Libya
Current position : Master program in biomedical science student
B. EDUCATIONAL BACKGROUND
No Name
ofSchool/Institution Location Degree Year
1 Alegwilat Alegwilat /Zliten , Libya Primary 2001
2 Hassan ben tabet aljoma /Zliten , Libya Secondary 2005
3 Zawia University Zawia , Libya Bachelor 2009
4 Diponegoro University Semarang,CentralJava,Ind
onesia Master
Program 2014
C. FAMILY HISTORY
1. Name of father : Mohamed RajabAljatlawi
2. Name of mother: FatmaAbdoallahAlter
ACKNOWLEDGEMENT
First of all, I would like to express my deepest sense of Gratitude to Diponegoro University for their support, encouragement and care. I would like to express my gratitude to all those who gave me the possibility to complete this thesis.
I am deeply grateful to :
1. Prof. Dr.dr. Tri Nur Kristina. DMM. M.Kes as Chairman of Program Administrator Master Biomedical Sciences, University of Diponegoro Semarang who has given an opportunity to gain knowledge in Biomedical Science Program Graduate Biomedical Sciences, University of DiponegoroSemarang.udiGraduateProgram who have given permission to continue (( his studies)) to such great degree.
2. My supervisor I: Prof.dr.LisyaniSuromo,Sp.PK(K) and Supervisor II:Dr.dr.RAKisdiaminatun.RMD.MSC whose help, stimulating suggestions and encouragement helped me in all the time of research and writing of this thesis.
3. My examiners Prof. Dr.dr.TriNur Kristina. DMM. M.Kes and Prof. dr. Edi Dharmana,M,Sc, PhD, Sp.Park.Thank you for the suggestions,ideas and comments for overall my project
vii
4. My former colleagues from the Department of Biomedicine Science of Diponegoro University in my class and the all staph. I want to thank them for all their help, support, interest and valuable hints.
5. Ms. NataSulastri, Mbakfika, Mas abdul who have helped smooth implemetation of administrative and facility study, I am really grateful to your help.
6. I want to thank the Department of biochemical at GadjahMada University for giving me permission to commence this thesis especially Prof.Dr.Wayan Tunas Artama.
7. I would like to thank my family my parents Mohamed RajebAljatalwi and my motherFatmaAbdoallah and my brothers,my sisters, for giving birth to me at the first place and supporting me spiritually throughout my life.
8. I would like to thank all of my friends who giving me support during my study and thanks for University of Diponegoro Semarang, biomedic department,kariadi hospital and Indonesia.
Semarang, May 30, 2014
SALEM.M.ALJATLAWI
TABLE OF CONTENT
Page
Title Page ... i
Approval Page... ii
Development Monitoring Sheet Result Research of Thesis ... iii
Declaration ... iv
Curriculum Vitae... v
Acknowledgement... . vi
Table of Content... viii
List of Obbreviation... xi
List of Figures ... xiv
List of Tables... xv
List of appendixs ... xvi
English Abstract ... xvii
Indonesia Abstrak... xvi
CHAPTER I. INTRODUCTION... 1
Background of study... 1
Research Questions... 4
Research Objective... 4
Research Benefits ... 4
Reserach Originality ... 5
ix
Page
CHAPTER II.LITERATURE REVIEW... 8
High fat diet... 8
Effect of high fat diet toward inflammation... 10
Main risk factor of a high fat diet... 11
Obesity... 11
Cardiovascular Disease... 12
Olive oil ... 13
Introduction... 13
Olive oil and its other benefits ... 15
Olive oil compounds which have important medical. ... 16
phenolic compounds. ... 16
polyphenols. ... 16
Oleic acid. ... 17
The effect of olive oil in inflammation... 17
Markers of Inflammation ... 18
CRP... 18
IL6 ... 20
CHAPTER III. THEORETICAL FRAMEWORK... 22
Theoretical Framework Literates ... 22
Theoretical Framework... 23
Conceptual Framework... 24
Hypothesis... 24
Major Hypothesis ... 24
Minor Hypothesis ... 24
CHAPTER IV.RESERACH METHOD... . 25
Scope of Study ... 25
Design of Study... 25
Population and Sample ... 26
Variables of Study ... 27
Operational Definiton ... 28
Research Instrument and Material... 28
Research Method... 29
Research Flow ... 30
Data Collection... 31
Data Analysis ... 31
Ethics ... 32
CHAPTER V. RESULTS... 33
The result of analyzed data ... 33
CHAPTER VI. DISCUSSION... 35
Limitation of study ... 37
CHAPTER VII. CONCLUSION, SUGGESTION, FURTHER RESEARCH... 38
References... 39
xi
LIST OF ABBREVIATION
BP : Blood pressure
BMI : Body mass index
Ca2 : Ionized calcium COX-1 : Cyclooxygenase-1
COX-2 : Cyclooxygenase-2
CRP : C-reactive protein
CVD : Cardiovascular disease
CHD : Coronary heart disease
DASH : Dietary Approaches to Stop Hypertension DAB : 3,3-diaminobenzidinetetrahydrochloride EVOO : Extra virgin olive oil
HDL : High-density lipoprotein
HFD : High fat diet
HsCRP : High sensitive C-reactive protein HSD : Honestly significant difference
K : Control group consist of control negative and control positive KOH : Potassium Hydroxide
LRP1 : Lipoprotein receptor-related protein MCP-1 : Monocyte chemoattractant protein MDA : Aortic malonyl dehydrogenase MUFA : Monounsaturated fatty acids
NFκB : Nuclear factor Κb
NaOH : Sodium Hydroxide
IL-6 : Interleukin 6 IFNγ : Interferon gamma IL-1 : Interleukin 1
IgG : Immunoglobulin
OLAB : Plasma autoantibodies against oxLDL OxLDL : Oxidized low density lipoprotein
OT1 : Outcome from treatment group I
OT2 : Outcome from treatment group II
OT3 : Outcome from treatment group III OA : Oleic acid
PARS : Peroxisome proliferatorsactivated receptor gamma PC : Phenolic compounds
R : Random allocation in 5 groups SMCs : Smooth muscle cells
TF : Tissue factor
TFPI : Tissue factor pathway inhibitor
TC : Cholesterol
TG : Triglyceride
TMD : Traditional Mediterranean diet
TNFα : Tumor necrosis factor
xiii
VOO : Virgin olive oil
WHO : World health organization
XT1 : Treatment group I ( high fat diet and olive oil dossage I) XT2 : Treatment group I ( high fat diet and olive oil dossage II) XT3 : Treatment group I ( high fat diet and olive oil dossage III)
LIST OF FIGURES
Page
Figure 1. Olive oil... 14
Figure 2. Theoretical Framework... 23
Figure 3. Conceptual Framework... 24
Figure 4.Desige of sudy... 25
Figure 5. Research Flow ... 30
xv
LIST OF TABLES
Page Table1. Previous report related to study on protective effect olive oil on high fat diet 6 Table 2. Operational definition ... 28 Table 3. IL-6 levels in HFD... 33 Table4. CRP levels in HFD... 34
LIST OF APPENDIXS
Page Appendix 1 . Statisticalanalysis ... 48 Appendix 2 . Ethical clearance... 63 Appendix 3 . Photo of the experiment... 65
xvii ABSTRACT
Background:High-fat diet is the most lethal habit which often leads to excess weight gain which increase the risk of obesity it may increase health problems.Substances found in extra-virgin olive oil has anti-inflammatory effects and the high concentration of mono-unsaturated fatty acids in olive oil may be beneficial for the heart.
Objective :To investigate the extra virgin olive oilhave an effect to IL6 and CRP in high fat diet wistar rats.
Method :Twenty five male Wistar rats were divided into the following five groups: rats on standard diet; rats on high fat diet; rats on high fat diet + EVOO 1ml/kg/day; rats on high fat diet +EVOO 2ml/kg/day; and rats on high fat diet + EVOO 3ml/kg/day. . After 8 weeks the rats were IL-6 and CRP levels were measured by means of ELISA.
Result:There was no significant different were observed between all groups for IL6 ( p = 0.755) and the test homogeneity of variances was non significant ( p = 0.637). There is no significant different were observed between all groups for CRP ( p = 0.939).
Conclusions:EVOO at any doses studied dosen’t reduce the IL-6 and CRP levels ofHFD rats.
Keywords :Extra virgin olive oil, high fat diet, IL-6, CRP, rats.
ABSTRAK
LATAR BELAKANG : Diet tinggi lemak adalah kebiasaan paling mematikan yang seringmenyebabkankelebihanberatbadandenganpeningkatanrisikoobesitas yang dapat meningkatkan masalah kesehatan. Zat yang ditemukan dalam ekstra alami minyak zaitun mempunyai efek anti-inflamasi dan konsentrasi asam lemak tak jenuh dalam minyak zaitun memiliki manfaat bagi jantung.
TUJUAN: Penelitian ini bertujuan untuk mengetahui ekstra alami minyak zaitun berpengaruh terhadap IL-6 dan CRP pada tikus Wistar yang diberi diet tinggi lemak.
METODE : 25 tikus Wistar jantan dibagi dalam 5 kelompok. Tikus dengan diet standar, tikus dengan diet tinggi lemak, tikus dengan diet tinggi lemak + EVOO 1 ml/kg/hari, tikus dengan diet tinggi lemak + EVOO 2 ml/kg/hari, tikus dengan diet tinggi lemak + EVOO 3 ml/kg/hari. Setelah 8 minggu tikus dibedah dan dilakukan pengukuran kadar IL-6 dan CRP dengan menggunakan ELISA.
HASIL : Tidak terdapat perbedaan signifikan rata-rata kadar IL-6 antara semua kelompok perlakuan (p=0,755) dan teshomogenitas tidak signifikan (p=0,637).
Tidak terdapat perbedaan signifikan rata-rata kadar CRP antara semua kelompok perlakuan (p=0,939).
SIMPULAN : Dalam penelitian ini, EVOO pada berbagai dosis tidak menurunkan kadar IL-6 dan CRP pada tikus Wistar yang diberi diet tinggilemak.
Kata kunci :Ekstra alami minyak zaitun, Diet Tinggi Lemak, IL-6, CRP.
