Транскрипційна індукція генів – регуляторів рециркуляції і хоумінгу лімфоцитів Madcam1, S1pr1, Cxcr4 і Ccr7 в умовах експериментального гестаційного діабету, що впливає на розподіл Т-клітин у брижових лімфатичних вузлах у нащадків

Abstract

Мета дослідження – з’ясувати рівень експресії мРНК генів Madcam1, S1pr1, Cxcr4 і CCR7 і його вплив на розподіл Th1-, Th17- і Treg-клітин у брижових лімфатичних вузлах у нащадків щурів з експериментальним гестаційним діабетом і за умов формування толерантності до орального введення інсуліну. Formation of peripheral immunological tolerance (PIT) to self-antigens is an important mechanism for preventing the development of autoimmune diseases. Maternal hyperglycemia that develops in gestational diabetes (GD) can influence on the morphogenesis of the immune system and leads to violations of PIT formation to pancreatic antigens. Using mucosa is an attractive way to treatment by administering antigens as tolerogen, especially in children. In animal models oral or intranasal administration of antigen can induce PIT. Mesenteric lymph nodes (MLN) is a major transition point for recirculating lymphocytes of gastrointestinal associated lymphoid tissue and at the same time - the main places for PIT induction. Homing of lymphocytes in MLN is regulated with adressin Madcam1, chemokine receptors Cxcr4 and Ccr7. And sphingosine-1-phosphate receptors S1pr1 activate T-cell exit from MLN. We studied descendants of intact Wistar rats (males), offspring of rats with experimental gestational diabetes (EGD) and descendants of rats with EGD which received short-acting human insulin orally using a pipette for the first 14 days of life at a dose of 30 IU. Each group was studied in age 1 and 6 months. We use RT-PCR method for investigating of mRNA expression levels of genes Madcam1, Cxcr4, Ccr7 and S1pr1 in MLN of experimental rats. As reference gene to determine the relative value of changes in the expression level of target genes was used glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene. Normalized relative quantity of cDNA target genes was determined by the method ΔΔCt. The structure of the population of T-bet+, Rorγt+, Foxp3+-cells (i.e., Th1, Th17 and Treg-lymphocytes) were studied based on the analysis of serial histological sections of MLN and data of their morphometric and densitometric characteristics. We determined the absolute (number of cells per 1 mm2 of area) and relative (%) density the subsets of immunopositive cells in the investigated areas of MLN.