Genska ekspresija enzima antioksidativne zaštite u tkivima pacova i humanoj krvi : uticaj spoljnih faktora i korelacija sa biohemijskim markerima oksidativnog stresa
Gene expression analyses of antioxidative enzymes in rats' tissues and human blood: the influence of external factors and correlation with biochemical markers of oxidative sress
Author
Ninić, Ana R.
Mentor
Spasojević-Kalimanovska, Vesna
Committee members
Spasić, SlavicaBogavac-Stanojević, Nataša

Kotur-Stevuljević, Jelena

Joiksić, Gordana

Metadata
Show full item recordAbstract
Genska ekspresija je proces koji obuhvata transkripciju gena, obradu transkripta i
njegovu translaciju u protein. Kvantifikacijom genske ekspresije može se videti da li
dolazi do ushodne ili nishodne regulacije specifičnih gena i posledica takve regulacije.
Značajnu ulogu u zaštiti od oksidativnog stresa (prekomerne količine stvorenih
slobodnih radikala) u intra- i ekstraćelijskom prostoru imaju enzimi antioksidativne
zaštite, superoksid-dismutaza (SOD) i paraoksonaza (PON). Interakcije gena
antioksidativne zaštite i fizičke aktivnosti, gena i tipa dijete, gena i terapije
hipolipemicima mogu da modifikuju ekspresiju tih gena; fizička aktivnost slobodnim
radikalima, aterogena dijeta i terapija atorvastatinom svojim komponentama, direktno ili
indirektno interagujući sa transkripcionim faktorima.
Cilj istraživanja je bio da se ispita uticaj fizičke aktivnosti na gensku ekspresiju
izoenzima SOD (Cu/Zn SOD i Mn SOD) u limfocitima sportista i uticaj aterogene dijete
i terapije atorvastatinom... na gensku ekspresiju izoenzima PON (PON1, PON2, PON3) u
jetri, belom masnom tkivu i aorti eksperimentalnih životinja. Pored toga, cilj je bio i da
se uporede metode za izolaciju RNK iz krvi i ispita njihov uticaj na vrednosti genske
ekspresije izoenzima SOD.
Istraživanje je obuhvatilo odrasle osobe (kontrolna grupa), grupu fudbalera pre i nakon
12 nedelja intenzivnog treniranja i eksperimentalne životinje koje su podeljene u 3
grupe (1. grupa na normalnoj dijeti – ND grupa, 2. grupa na aterogenoj dijeti - AD i 3.
grupa na aterogenoj dijeti i terapiji atorvastatinom - ADA grupa). RNK je izolovana
TRIzol™ metodom (TRI) iz mononuklearnih ćelija periferne krvi (TRIMĆPK) i na
aparatu ABI Prism™ 6100 Nucleic Acid PrepStation (ABI) iz krvi (ABIKRV) i
mononuklearnih ćelija periferne krvi (ABIMĆPK). Takođe, RNK je izolovana pomoću
TRIzol™ metode iz ispitivanih organa pacova. Za kvantifikaciju relativne genske
ekspresije korišćena je Real time PCR metoda.
Fudbaleri su nakon 12 nedelja treniranja bili u oksidativnom stresu. Vrednosti genske
ekspresije Cu/Zn SOD se nisu statistički značajno promenile (p=0,106), dok su
vrednosti Mn SOD bile značajno veće (p=0,031) nakon treniranja. Vrednosti genske ekspresije Cu/Zn SOD su bile parametar koji nezavisno utiče na ukupnu aktivnost SOD
u plazmi (p=0,025), a koncentracija tiobarbituratna kiselina-reagujućih supstanci je
imala ulogu prediktivnog faktora (p=0,017) za vrednosti genske ekspresije Mn SOD...
Gene expression is a process that includes transcription, transcript processing and its
translation into protein. Gene expression quantification can be used to see weather
specific genes are up or down regulated and consequences of such regulation. Enzymes
of antioxidative defense system, superoxide dismutase (SOD) and paraoxonase (PON)
have important roles in protection against oxidative stress (excessive amount of
generated free radicals) in intra- and extracellular space. Interaction of antioxidative
defense genes and physical activity, genes and type of diet, genes and hipolipemic drugs
can modify the expression patterns of these genes; physical activity by free radicals
formation, atherogenic diet and atorvastatin therapy with its components, directly or
indirectly interacting with transcription factors.
The aim of this study was to investigate the influence of physical activity on SOD
isoenzymes’ (Cu/Zn SOD and Mn SOD) gene expression in sportsmen’ lymphocytes
and the influence o...f atherogenic diet and atorvastatin therapy on paraoxonase
isoenzymes’ (PON1, PON2 and PON3) gene expression in liver, white adipose tissue
and aorta of experimental animals. In addition, we compared methods for RNA
isolation from blood and examined their influence on SOD isoenzymes’ gene
expression.
The study included control group, soccer players before and after 12 weeks of intensive
trainings and rats who were divided into 3 groups (1. group on normal diet - ND group,
2. group on atherogenic diet - AD group and 3. group on atherogenic diet with
atorvastatin therapy- ADA group). RNA was isolated using Trizol™ method (TRI) from
peripheral blood mononuclear cells (TRIPBMC) and using ABI Prism ™ 6100 Nucleic
Acid PrepStation (ABI) from blood (TRIBLOOD) and from peripheral blood mononuclear
cells (ABIPBMC). RNA was also isolated from rats’ organs using TRI method. Real time
PCR was used for relative gene expression quantification.
Soccer players were in oxidative stress after 12 weeks of intensive trainings. Cu/Zn
SOD gene expression levels did not change significantly after training (p=0.106), while
those of Mn SOD were significantly higher (p=0.031) after than before training. Cu/Zn SOD expression levels were independent parameter that influenced total SOD activity
in plasma (p=0.025). Concentration of thiobarbituric acid reactive substances was
predictive parameter for Mn SOD gene expression values (p=0.017)...