Features of immune metabolism of lymphocytes in pancreatic lymph nodes during experimental steptozotocin-induced diabetes mellitus and after introduction of metformin.

Autor: D. A. Putilin, A. M. Kamyshny
Jazyk: Russian<br />Ukrainian
Rok vydání: 2016
Předmět:
Zdroj: Морфологія, Vol 10, Iss 2, Pp 61-68 (2016)
Druh dokumentu: article
ISSN: 1997-9665
DOI: 10.26641/1997-9665.2016.2.61-68
Popis: Background. Metabolic changes in terms of developing diabetes, especially hyperglycemia, can directly affect immune metabolism of lymphocytes. Objective: The aim was to find out the level of mRNA gene expression of Glut1, mTOR and AMPK1α in pancreatic lymphatic nodes (PLN) in rats with experimental streptozotocin-induced diabetes mellitus (ESIDM) and after introduction of metformin. Methods. Glut1, mTOR and AMPK1α mRNA expression were analyzed by real-time reverse transcriptase-polymerase chain reaction. Total RNA was extracted from PLN tissue by Trizol RNA Prep 100 (Isogen, Russia), according to the manufacturer's instructions. RNA was re-suspended in RNase free water, quantified and subjected to RT-PCR reaction using RT-PCR kit; RT (Syntol, Russia). To determine the level of Glut1, mTOR and AMPK1α mRNA, RT-PCR was performed in real-time by thermal cycler CFX96 ™ Real-Time PCR Detection Systems (Bio-Rad Laboratories, Inc., USA). The relative level of gene expression were studied with rat reference genes GAPDH by the method ΔΔCt. Statistical analysis were conducted using available software «Bio-Rad SFX Manager 3.1» (Bio-Rad, USA). Results. It was established that hyperglycemia caused the transcript induction of genes of glucose transporter Glut1 (in 9,9-28,9 times, p
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