The Effect of TLR Agonists and Myokines on Secretory Activity of Adipogenically Differentiated MSC Cultures.

We studied the effect of bacterial pathogen-associated molecular patterns and myokines on the secretion of adipokines by mesenchymal stem cells (MSC) and products of their adipogenic differentiation. The secretion of adiponectin, adipsin, leptin, and insulin by adipogenically differentiated cell cultures was quantitatively determined using multiplex ELISA. MSC obtained from the stromal vascular fraction of human subcutaneous adipose tissue were shown to secrete a known adipokine adipsin. The ability of white adipocytes to secrete significant amounts of insulin (in vitro) has been shown for the first time. Control cultures of white adipocytes secreted much higher levels of adiponectin, leptin, and insulin when compared to other adipocytes cultures. On the other hand, beige and brown adipocyte cultures secreted more adipsin than white adipocyte cultures. The influence of myokine ß-aminoisobutyric acid on the secretion of adipsin in MSC, white, beige, and brown adipocytes was also studied.

The Effect of LPS and Flagellin on the Process of Lipolysis in Mesenchymal Stromal Cells during Adipogenic Differentiation.

We analyzed the effects of bacterial pathogen-associated molecular patterns (LPS and flagellin) and adrenergic agonist isoproterenol on the content of total and phosphorylated (Ser552) hormone-sensitive lipase in mesenchymal stromal cells and cell products of their adipogenic differentiation. The expression of hormone-sensitive lipase and an increase in the content of its activated phosphorylated form were demonstrated by Western blotting in cells of all three lines of adipogenic differentiation. Under the influence of flagellin, the content of total and phosphorylated forms of hormone-sensitive lipase increased in brown adipocytes, while LPS induced a decrease in the content of total hormone-sensitive lipase in white adipocytes. We hypothesize that bacterial pathogen-associated molecular patterns can activate lipolysis under pathological conditions associated with slow remodeling of the adipose tissue.

[The study of tryptophan metabolite concentrations in blood serum and fecal extracts from obese children]. / Izuchenie soderzhaniia metabolitov obmena triptofana v syvorotke krovi i kalovykh ékstraktakh u detei s ozhireniem.

We found that changes in the concentrations of tryptophan metabolites in the blood serum and in the intestinal contents are one of the mechanisms for the formation of metabolic coupling in the system "macroorganism-intestinal microbiota", which undergoes significant changes in the development of obesity. Although blood kynurenine remained basically unchanged in obese children we found an increase in some of its serum metabolites: anthranilic, kynurenic and xanthurenic acids. It is noteworthy that in the analysis of fecal matter in obese children, revealed a 2-fold increase in the level of kynurenine while the concentration of kynurenine pathway metabolites corresponded to the level of the group of healthy children. This may indicate the metabolic activation of the microbiota associated with the intestinal mucosa. This is also supported by the absence of statistically significant differences in the concentration of indole in healthy children and in obese children in fecal analyses, and a significant increase in the concentration of indole-3-lactate and indole-3-acetate in the blood serum of obese children.

[Humoral regulation of nutritional status in children].

The purpose of the research was the investigation of the characteristics of insulin resistance, levels of adipokines, myokines and collagen metabolites type I in children in orphanages and social rehabilitation centers (SRC). The study involved 69 children aged 7-12 years. The first group consisted of 20 children from SRC, the second - 16 children living in orphanages, control group included 33 children (1-2 health groups) from wealthy families. The average length of stay of children in SRP was 1.0 (0.5-2.5) month, in orphanages - 38 (22-44) months. The groups were comparable in age and sex. Anthropometric indices were determined, in blood serum glucose concentration was measured by glucose oxidase method, blood serum level of insulin, cortisol, leptin, adiponectin, resistin, apelin, myostatin, collagen metabolites were determined by ELISA. Children from orphanages and SRC had significantly lower anthropometric indices in comparison with the control group. Particularly low parameters were observed in children from SRP, which in growth, body weight, chest circumference and body mass index significantly lagged behind the control group peers. The children from SRP as compared with the control group revealed higher insulin resistance index (3.33±0.49 vs 2.29±0.29, p<0.05) and cortisol level (405.21±38.21 vs 313.08±25.97 nmol/l, p>0.05). There were changes (p<0.05) in blood serum content of adipokines - reduced leptin level (5.35±1.55 vs 14.00±3.10 ng/ml) and increased apelin level (4.07±0.82 vs 2.19±0.41 ng/ml), as well as higher levels (p<0.05) of myostatin (183.95±16.3 vs 116.4±9.4 ng/ml) and C-terminal telopeptide of type I collagen (1.11±0.11 vs 0.72±0.08 ng/ml). The children in orphanages compared with the control group had lower levels of insulin (5.04±0.69 vs 11.3±1.12 µIU/ml), high insulin sensitivity - HOMA-IR was 1.01±0.16. The children in orphanages have even more pronounced reduction in leptin concentration (2.69±0.4 ng/ml), a decrease of resistin (4.99±0.32 vs 7.16±0.70 ng/ml, p<0.05), increased concentration (p<0.05) of apelin (3.53±0.67 ng/ml), myostatin (181.17±10.2 ng/ml) and C-terminal telopeptide of type I collagen (2.70±0.22 ng/ml). Features of hormone and cytokine regulation of metabolism make it particularly urgent to prevent disorders of eating behavior and to promote a healthy lifestyle among children left without parental care.

Study of genetic stability of human bone marrow multipotent mesenchymal stromal cells.

Immunophenotype, proliferation rate, and genetic stability parameters of bone marrow multipotent mesenchymal stromal cells were studied. Despite the reduction of proliferative activity by passages 11-12, the cells retained the characteristic immunophenotype. The incidence of spontaneous aneuploidy for autosomes 6, 8, 11 and sex chromosomes was evaluated. Two cultures of mesenchymal stromal cells carrying aneuploid cell clones were detected: with chromosome 8 trisomy and X chromosome monosomy. The results indicate the possibility of genetic transformation and selection of mesenchymal stromal cells with abnormal karyotype during in vitro culturing.