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Statements

Subject Item
n2:RIV%2F00216208%3A11160%2F14%3A10281587%21RIV15-MSM-11160___
rdf:type
n13:Vysledek skos:Concept
rdfs:seeAlso
http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086033
dcterms:description
Obesity and metabolic syndrome is increasing health problem worldwide. Among other ways, nutritional intervention using phytochemicals is important method for treatment and prevention of this disease. Recent studies have shown that certain phytochemicals could alter the expression of specific genes and microRNAs (miRNAs) that play a fundamental role in the pathogenesis of obesity. For study of the obesity and its treatment, monosodium glutamate (MSG)-injected mice with developed central obesity, insulin resistance and liver lipid accumulation are frequently used animal models. To understand the mechanism of phytochemicals action in obese animals, the study of selected genes expression together with miRNA quantification is extremely important. For this purpose, real-time quantitative PCR is a sensitive and reproducible method, but it depends on proper normalization entirely. The aim of present study was to identify the appropriate reference genes for mRNA and miRNA quantification in MSG mice treated with green tea catechins, potential anti-obesity phytochemicals. Two sets of reference genes were tested: first set contained seven commonly used genes for normalization of messenger RNA, the second set of candidate reference genes included ten small RNAs for normalization of miRNA. The expression stability of these reference genes were tested upon treatment of mice with catechins using geNorm, NormFinder and BestKeeper algorithms. Selected normalizers for mRNA quantification were tested and validated on expression of NAD(P)H:quinone oxidoreductase, biotransformation enzyme known to be modified by catechins. The effect of selected normalizers for miRNA quantification was tested on two obesity- and diabetes- related miRNAs, miR-221 and miR-29b, respectively. Finally, the combinations of B2M/18S/HPRT1 and miR-16/sno234 were validated as optimal reference genes for mRNA and miRNA quantification in liver and 18S/RPlP0/HPRT1 and sno234/miR-186 in small intestine of MSG mice. Obesity and metabolic syndrome is increasing health problem worldwide. Among other ways, nutritional intervention using phytochemicals is important method for treatment and prevention of this disease. Recent studies have shown that certain phytochemicals could alter the expression of specific genes and microRNAs (miRNAs) that play a fundamental role in the pathogenesis of obesity. For study of the obesity and its treatment, monosodium glutamate (MSG)-injected mice with developed central obesity, insulin resistance and liver lipid accumulation are frequently used animal models. To understand the mechanism of phytochemicals action in obese animals, the study of selected genes expression together with miRNA quantification is extremely important. For this purpose, real-time quantitative PCR is a sensitive and reproducible method, but it depends on proper normalization entirely. The aim of present study was to identify the appropriate reference genes for mRNA and miRNA quantification in MSG mice treated with green tea catechins, potential anti-obesity phytochemicals. Two sets of reference genes were tested: first set contained seven commonly used genes for normalization of messenger RNA, the second set of candidate reference genes included ten small RNAs for normalization of miRNA. The expression stability of these reference genes were tested upon treatment of mice with catechins using geNorm, NormFinder and BestKeeper algorithms. Selected normalizers for mRNA quantification were tested and validated on expression of NAD(P)H:quinone oxidoreductase, biotransformation enzyme known to be modified by catechins. The effect of selected normalizers for miRNA quantification was tested on two obesity- and diabetes- related miRNAs, miR-221 and miR-29b, respectively. Finally, the combinations of B2M/18S/HPRT1 and miR-16/sno234 were validated as optimal reference genes for mRNA and miRNA quantification in liver and 18S/RPlP0/HPRT1 and sno234/miR-186 in small intestine of MSG mice.
dcterms:title
Reference genes for real-time PCR quantification of messenger RNAs and microRNAs in mouse model of obesity Reference genes for real-time PCR quantification of messenger RNAs and microRNAs in mouse model of obesity
skos:prefLabel
Reference genes for real-time PCR quantification of messenger RNAs and microRNAs in mouse model of obesity Reference genes for real-time PCR quantification of messenger RNAs and microRNAs in mouse model of obesity
skos:notation
RIV/00216208:11160/14:10281587!RIV15-MSM-11160___
n3:aktivita
n12:P n12:I
n3:aktivity
I, P(EE2.3.30.0022), P(GBP303/12/G163)
n3:cisloPeriodika
1
n3:dodaniDat
n17:2015
n3:domaciTvurceVysledku
n9:1405764 n9:8996563 n9:2442256 n9:9958185 n9:2270455 n9:1935240
n3:druhVysledku
n14:J
n3:duvernostUdaju
n15:S
n3:entitaPredkladatele
n20:predkladatel
n3:idSjednocenehoVysledku
41742
n3:idVysledku
RIV/00216208:11160/14:10281587
n3:jazykVysledku
n8:eng
n3:klicovaSlova
qPCR; normalization; monosodium glutamate obesity; miRNA; gene expression; catechin
n3:klicoveSlovo
n6:miRNA n6:qPCR n6:monosodium%20glutamate%20obesity n6:catechin n6:normalization n6:gene%20expression
n3:kodStatuVydavatele
US - Spojené státy americké
n3:kontrolniKodProRIV
[0023E8DD05BA]
n3:nazevZdroje
PLoS ONE
n3:obor
n18:FR
n3:pocetDomacichTvurcuVysledku
6
n3:pocetTvurcuVysledku
6
n3:projekt
n5:GBP303%2F12%2FG163 n5:EE2.3.30.0022
n3:rokUplatneniVysledku
n17:2014
n3:svazekPeriodika
9
n3:tvurceVysledku
Szotáková, Barbora Bártíková, Hana Skálová, Lenka Matoušková, Petra Hanušová, Veronika Boušová, Iva
n3:wos
000330237000085
s:issn
1932-6203
s:numberOfPages
11
n7:doi
10.1371/journal.pone.0086033
n19:organizacniJednotka
11160