Effects of miR-146a on hepatic INSR gene expression in F1 offspring of paternal rats with high-glucose-high-fat diet
10.3760/cma.j.issn.1000-6699.2019.05.010
- VernacularTitle:miR-146a对父代高脂高糖饮食背景下子代新生鼠肝脏胰岛素受体基因表达的影响
- Author:
Hua TANG
1
;
Wei DANG
;
Yingli WEI
;
Yinli CAO
Author Information
1. 空军军医大学第二附属医院消化内科
- Keywords:
miR-146a;
Methylation;
Insulin receptor;
High-glucose-high-fat diet;
Male;
Sprague Dawley rats
- From:
Chinese Journal of Endocrinology and Metabolism
2019;35(5):404-409
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effect of miR-146a on hepatic INSR gene expression in F1 offspring of paternal rats with high-glucose-high-fat diet ( HGFD) . Methods 5-week-old male SD rats were randomly divided into normal diet ( ND) and HGFD groups. Male rats with ND or HGFD feeding for three months mated with normal female ones. Blood glucose concentration, glucose tolerance, and insulin tolerance in newborn male offspring rats were detected respectively. Differential miRNAs between ND and HGFD groups were compared using next generation sequencing and were then confirmed by real-time quantitative PCR ( qPCR) . The relative expression of INSR mRNA and the methylation level of INSR promoter in liver tissues of the offspring newborn rat were detected and the correlations between them and the relative expression of differential microRNA were analyzed respectively. In vitro, effects of miR-146a on expression and methylation of INSR gene in BRL-3A cells were detected using Western-blot assay and qPCR respectively. Results The fasting glucose concentration of different groups were without significant difference, but glucose tolerance and insulin tolerance of neonatal male rats in HGFD group were decreased significantly . Next generation sequencing has revealed 45 up-regulated miRNAs and 15 down-regulated miRNAs in HGFD group. Among them, differences of 8 miRNAs expression in the enlarged samples were confirmed by qPCR. miR-146a was up-regulated for more than 10 times in the liver of the offspring of HGFD group. Expression level of miRNA-146a was negatively correlated with the relative expression of INSR and positively correlated with the methylation level of INSR in livers of neonatal rats in HGFD group. In vitro, miR-146a ( mimics ) promoted the methylation of INSR gene and inhibited the expression of INSR in BRL-3A cells. Conclusion HGFD feeding to male SD rats leads to the inhibition of hepatic INSR gene expression in neonatal offspring via upregulating miR-146a.
