Document Type : Original Article (s)
Authors
1
MSc Student, Department of Genetics and Molecular Biology, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran
2
Department of Medical Biotechnology, School of Medicine, Arak University of Medical Sciences, Arak, Iran
3
Department of Biology, School of Sciences, University of Isfahan, Isfahan, Iran
4
Associate Professor, Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
Abstract
Background: Type 2 diabetes (T2D) is the most prevalent chronic metabolic disease. MicroRNAs act as regulators of gene expression by inhibiting translation or promoting degradation of target mRNAs. Recent reports indicate that microRNAs can play important role in establishment of diabetes and has roles in type 2 diabetes different aspects including insulin insensitivity. To detect a noninvasive screening method for type 2 diabetes, we evaluated the expression level of miR-103 in peripheral blood mononuclear cells (PBMC) samples from pre-diabetes and type 2 diabetes rats in compare to normal group (n = 8 in each group) using real-time polymerase chain reaction (RT-PCR) method.Methods: Animal model type 2 diabetes was made with combination of high-fat diet (HFD) and low dose of stretozotocin (STZ) injection. Blood glucose level, serum insulin, and cholesterol, low density lipid and triglyceride levels were measured after 10-weeks diet interference. Peripheral blood mononuclear cells isolated from blood samples of three groups of animals, and then total RNA was obtained using Trizol reagent. All microRNAs converted to cDNA (complementary DNA). Expression level of rno-miR-103 was assessed via one-step SYBR GreenⅠ relative real-time polymerase chain reaction, using specific LNA TM qRT-PCR primer set for amplifying rno-miR-103. Statistical analysis was performed using SPSS20 software.Findings: Combination of high-fat diet and low dose of stretozotocin effectively induced type 2 diabetes in rats. rno-miR-103 expression level in peripheral blood mononuclear cells samples of pre-diabetic (high-fat diet) subjects and type 2 diabetes rats significantly elevated in compare to normal group.Conclusion: The high-fat diet/stretozotocin-induced type 2 diabetes animal model is the most suitable way for mimicking pre-diabetes and diabetes in men. The result of miRNA expression assay in peripheral blood mononuclear cells indicates the importance of rno-miR-103 in type 2 diabetes pathogenesis in early stages and has potential as a clinically useful noninvasive source to predict the disease status.
Keywords