The effects of interleukin-10 and fibroblasts growth factor-2 in mice with toxic cuprizone model of demyelination

Abstract

Cytokines and growth factors exhibit neurotropic, anti-inflammatory and immunomodulatory properties, and therefore can affect the functioning of the nervous system at demyelinating disorders.Purpose. To identify changes in T-lymphocytes, macrophages, oxidative stress and antioxidant defence factors, endocrine thymus function in the brain and behaviour of mice receiving neurotoxin cuprizone and recombinant human proteins: interleukin-10 (rhIL-10) and fibroblast growth factor (rhFGF-2).Materials and methods. 4-6-month-old 129/Sv mice received cuprizone with food every day for 3 weeks. From the 7th day of cuprizone diet, they received different doses of rhIL-10 and rhFGF-2. The content of СD3+ Т-cells, macrophages, malondialdehyde, activity of antioxidant enzymes in the brain and the level of thymulin in the blood were determined. Behavioural reactions were assessed in the “open field” test.Results. In the brain of mice receiving cuprizone and rhIL-10, there was a decrease in the number of СD3+ Т-cells and the activity of macrophages, which significantly increased under the influence of the toxin. After cytokine injection, the activity of superoxide dismutase, catalase and glutathione peroxidase increased significantly in the brain, as well as the level of thymulin in the blood. Motor, emotional and exploratory activity of mice, significantly suppressed by the cuprizone, was increased after the introduction of rhIL-10. The effect of rhIL-10 on the test parameters is more pronounced at a dose of 5 μg/kg than 50 μg/kg. After injection of rhFGF-2 in the mice with cuprizone diet, there is a decrease in the activity of brain macrophages and an increase of the level of thymulin in the blood depending on the dose of this factor; the motor activity of these animals increased regardless of the rhFGF-2 dose.Conclusion. The injections of rhIL-10 and rhFGF-2 provide dose-dependent positive effects on the pathogenetic factors of experimental demyelinating pathology, as well as the functional state of the nervous system. Whereas, the injections of rhIL-10 have more pronounced effects.

The effects of combined administration of human umbilical cord-derived multipotent mesenchymal stromal cells and melatonin or fibroblast growth factor-2 to aged mice with a toxic cuprizone model of demyelination

“…RhFGF-2, dissolved in saline, was administered to mice intraperitoneally at a dose of 20 μg/kg, starting from the 11 th day of the cuprizone diet, with an interval of 48 hours, a total of 7 injections. This growth factor administration scheme has shown its effectiveness in our previous studies on adult mice with a cuprizone model of demyelination [21].…”

“…Fibroblast growth factor-2 (FGF-2, basic FGF) alters the migration, proliferation, and differentiation of MMSCs [17,18]. This growth factor also reduces the number of microglial cells and macrophages in the brain, changes the activity of antioxidant enzymes, and improves behavioral responses in animals with experimental models of multiple sclerosis [19][20][21][22]. The production of FGF-2 in MMSCs changes under the influence of melatonin [23].…”

“…Recombinant human fibroblast growth factor-2 (rhFGF-2) expressed in E. coli cells according to the standard procedure [21]. After the lysis of bacterial cells, the resulting protein was purified by affinity chromatography using Heparin-Sepharose to a homogeneity (not less than 98 %).…”

“…The factors of oxidative stress and antioxidant protection of the brain. The level of malondialdehyde (MDA) in the brain of mice was measured using a μQuant spectrophotometer (BioTek, USA) at a wavelength of 535 nm by the intensity of staining of the trimethine complex, which is formed between MDA and thiobarbituric acid, as described previously [10,21].…”

“…The activity of antioxidant enzymes was measured in supernatants of brain homogenates by the spectrophotometric method on a μQuant spectrophotometer (BioTek, USA), as we described earlier [10,21]. Superoxide dismutase (SOD) activity was measured in conventional units according to its ability to inhibit the reaction of adrenaline autooxidation into adrenochrome at pH 10.2 per 1 mg of protein per 1 min; catalase activity -in micromoles of utilized Н 2О2 per 1 mg of protein per 1 min; activity of glutathione peroxidase (GP) and glutathione reductase (GR) -in nanomoles of oxidized NADPH per 1 mg of protein per 1 min.…”