Immunophenotypical aspects of lung and spleen macrophages derived animals with the model of alloxan diabetes (type I) and their correction by sodium aminodigydrophtalazindione in vitro

Macrophages are found in all tissues and organs and display functional plasticity, which is necessary to maintain homeostasis, tissue regeneration and immunity. The macrophage phenotype is determined by microenvironment signals. Macrophages are traditionally classified into subsets- such as classically (M1) or alternatively (M2) activated macrophages. In the pathogenesis of diabetes mellitus (T1DM), M1 macrophages contribute to damage to the islets of Langerhans, loss of β-cells, causing autophagy, which can result in development of persistent infection increasing the risk of death from influenza or pneumonia in patients with type 1 diabetes. Therefore, it seems Important to study functional response of resident macrophages in organs and tissues not targeted in development of diabetes mellitus, as well as in response to ADPH stimulation that showed modulatory effect on immunocompetent cells.
In this study morphological and functional characteristics of macrophage cell cultures obtained from different sites in intact animal (IA) and modeled type 1 diabetes mellitus (DM1) were investigated. For this, we examined macrophage cell cultures isolated from rat liver and peritoneal cavity to be stimulated in vitro for 24 and 72 hours with a sodium aminodigydrophtalazindione. Cells, nucleus, cytoplasm area were measured and nuclear cytoplasmic ratio (NCR) were calculated. The phenotype was determined by surface expression of CD163 (M2-macrophages) and CD80 (M1-macrophages) receptors. Macrophage cytokine activity was determined by measuring IL-1α, IL-10 и TNFα level. ADPH effects on animal macrophages with DM1 after 24 h of exposure also led to a changedmorphometric parameters (decreased size of the nucleus and cells of the spleen macrophages, increased size of the nucleus of the alveolar macrophages, increased NCR in spleen macrophages) and production activity of the cells (increased levels of IL-1α and TNF α in almost all cell populations). After 72 h of cultivation, the levels of IL-1α and TNFα decreased in alveolar macrophages, splenic macrophages, whereas TNFα level was decreased, but IL-1α asmount was increased. The expression of surface cell markers for M1 and M2 phenotypes was also affected by ADPH so that CD163 expression was increased in stimulated alveolar macrophages isolated from animals with type 1 diabetes.