To investigate the effects of the rhLIF recombinant human leukemia inhibitory factor on the structural and functional changes in the central nervous system in the cuprizone-induced experimental model of multiple sclerosis in mice and to evaluate the involvement of the brain macrophages, T-lymphocytes and antioxidant enzymes in the observed rhLIF effects. Methods. Young 129/Sv mice were fed with the cuprizone neurotoxin daily for three weeks. rhLIF was injected daily (50 μg/kg) after seven days of the cuprizone treatment. We used histology, flow cytometry, spectrophotometric and functional methods. Results. The cuprizone treatment resulted in an increase in the number of the brain and spinal cord neurons with destructive changes, the brain macrophages, CD3 +-cells and the content of malondialdehyde; it changes the behavior of animals, decreases the brain superoxide dismutase and glutathione peroxidase activities. The rhLIF injection partially or completely restored the cuprizone-changed parameters. The effect of rhLIF was preserved during two months after the experiment completion. Conclusions. The neuroprotective effect of rhLIF is connected with a decrease in the cuprizone-induced changes in the number of the brain T-cells and macrophages and the activity of antioxidant enzymes.

The search for tools that increase the effectiveness of cell therapy of demyelinating pathology is relevant. They may be preparations that affect the pathogenetic factors of this pathology, in particular, the pineal hormone melatonin.The purpose of the work is to evaluate the involvement of immune system and antioxidant defense in the implementation of the protective effects of melatonin on morpho-functional disorders in the central nervous system induced by neurotoxin cuprizone.Materials and methods. The toxic demyelination model was induced on 129/Sv mice at the age of 3-5 months by adding cuprizone to food for 3 weeks. Since the 7th day of cuprizone administration, melatonin was injected intraperitoneally at 18:00 daily, at a dose of 1 mg/kg. In the brain of mice, the proportion of CD3+, Nestin+ cells and phagocytic macrophages, the content of malondialdehyde and the activity of antioxidant enzymes was determined. Blood serum was tested for thymic hormone thymulin levels. In the animals, we evaluated the structure of the brain and spinal cord neurons by Nissl staining of histological sections as well as analyzed behavioural reactions in the "open field" test.Results. In the brain of the mice received cuprizone, the proportion of CD3+ and Nestin+ cells, active macrophages and malondialdehyde content increases, glutathione peroxidase and glutathione reductase levels decreases. In the brain and spinal cord of the mice with a cuprizone diet, the proportion of altered neurons increases, and motor and emotional activity decreases. The introduction of melatonin results in a decrease in the relative number of CD3+ cells, active macrophages and malondialdehyde content, increased activity of glutathione peroxidase, glutathione reductase and level of thymulin. In such mice, the proportion of unchanged neurons increases as the number of Nestin+ cells decreases and behavioural responses are also improved.Conclusions. The neuroprotective effect of melatonin in demyelinating pathology is realized through the factors of the immune system and oxidative stress. The results may be useful in the development of new biotechnological approaches to the treatment of demyelinating pathology, in particular, multiple sclerosis.

No abstract

На сьогодні в Україні зареєстровано 8 комбінованих лікарських засоби кропиви собачої, настойку випускають 10 вітчизняних підприємств, фасовану сировину -3 вітчизняні заводи. Настойка собачої кропиви має певні недоліки: непостійність хімічного складу, і відповідно, фармакодинаміки; містить етиловий спирт, тому протипоказана дітям, вагітним, особам, діяльність яких потребує підвищеної уваги тощо. У зв'язку з цим, розробка стандартизованих лікарських засобів на основі настойки трави собачої кропиви, яка б не містила в своєму складі етанол, є актуальним завданням. Мета. Визначити параметри стандартизації модифікованого сухого екстракту з настойки собачої кропиви та розробити проект методик контролю якості на цю субстанцію. Методи дослідження. Об'єктом дослідження був сухий екстракт на основі собачої кропиви настойки (Виробник: ОАО «Лубнифарм», серія 062021), який був модифікований з лізином. Для аналізу використовували три серії екстрактів. При визначенні параметрів стандартизації одержаного сухого екстракту собачої кропиви трави використовували стандартні фармакопейні методики. За основу розробки методик контролю якості на сухий екстракт собачої кропиви були взяті монографії ДФУ «Собачої кропиви трава», «Собачої кропиви настойка» та «Лізину гідрохлорид». Результати дослідження. Визначено параметри стандартизації модифікованого сухого екстракту собачої кропиви трави. Запропоновано контролювати якість одержаного екстракту за такими показниками: описом, розчинністю, ідентифікувати екстракт методом ТШХ за вмістом флавоноїдів, іридоїдів та лізину, втратою в масі при висушуванні, залишковою кількістю органічних розчинників (етанолу), мікробіологічною чистотою, вмістом важких металів, кількісну стандартизацію проводити за вмістом флавоноїдів (не менше 2 %) та іридоїдів (не менше 0,5 %). Проаналізовано три серії екстракту згідно запропонованого проекту МКЯ, які повністю відповідали йому за всіма показниками. Висновки. Визначено параметри стандартизації модифікованого сухого екстракту з настойки собачої кропиви, який був одержаний з використанням лізину, та розроблено проект методик контролю якості на цю субстанцію. Проаналізовано три серії екстракту згідно запропонованого проекту МКЯ, які повністю відповідали йому за всіма показниками. Ключові слова: собача кропива, трава, настойка, модифікація, лізин, сухий екстракт, біологічно активні речовини, параметри, стандартизація.

The influence of histamine and quercetin, as well as their combined effect on the content of histamine in whole blood, the content of the superoxide anion radical, sialic acids, sulfhydryl groups, the activity of catalase, glutathione peroxidase, and glutathione transferase in rat erythrocytes was studied. It was established that quercetin at a concentration of 0.1 mM causes an increase in the content of histamine in the whole blood of rats, while all other studied concentrations cause a significant decrease in the content of biogenic amine. Adding histamine to whole blood at a concentration of 1 μM leads to a decrease in the content of endogenous histamine in the blood, while at a concentration of 10 μM, it causes an increase in the content of this biogenic amine. The combined effect of exogenous histamine and quercetin mainly leads to an increase in the amount of endogenous histamine in the whole blood of rats. In rat erythrocytes, quercetin causes the generation of the superoxide anion radical. An increase in the superoxide anion radical content occurs under the influence of histamine at a concentration of 0.1; 1 and 10 μM, while under the action of biogenic amine 0.01 μM, the amount of the studied product decreases. The combined effect of histamine and quercetin intensifies the formation of superoxide anion radical in erythrocytes, in addition to the effect of flavonoid in therapeutic concentration. Adding quercetin and histamine to whole blood causes an increase in the content of sialic acids. Such an effect was also found under the combined action of histamine at a concentration of 0.01 µM and quercetin at a concentration of 0.1; 0.5; 3; 5 mM. The combined effect of histamine at a concentration of 10 µM and quercetin at a concentration of 5 mM leads to a decrease in the content of sialic acids in erythrocytes. Addition of quercetin to whole blood causes an increase in the content of sulfhydryl groups, except for a concentration of 5 mM, at which the content of this indicator decreases. Histamine in a concentration of 0.01; 0.1 μM leads to an increase in the content of SH-groups, and at a concentration of 1 μM – to a decrease. Histamine against the background of exposure to quercetin leads to an increase in the content of sulfhydryl groups. The degree of influence of histamine and quercetin, as well as their combined effect, is the same on the content of endogenous histamine in the blood, sialic acids, sulfhydryl groups in erythrocytes. The independent effect of histamine and quercetin causes a weak effect on the indicated indicators of blood and erythrocytes. The combined effect of histamine and quercetin on the content of the studied indicators has a more pronounced effect. The most significant effect of the combined effect of histamine and quercetin was found on the content of the superoxide anion radical and sialic acids. Quercetin in a concentration of 0.1; 3; 5 mM leads to a decrease in catalase activity, while the studied flavonoid, with a concentration of 0.3; 1 mM, leads to an increase in the activity of the studied enzyme. Histamine at a concentration of 0.1 and 10 µM activates catalase, while biogenic amine (at a concentration of 0.01 and 1 µM) reduces the activity of the enzyme. The combined action of histamine and quercetin leads to an increase in the activity of catalase in hemolysates of erythrocytes of rats. Quercetin only at a concentration of 5 mM, histamine at a concentration of 0.1; 1; 10 μM enhances the activity of glutathione peroxidase. Simultaneous addition to the blood of histamine at a concentration of 10 μM and quercetin at a concentration of 3 and 5 mM, as well as the combined effect of histamine at a concentration of 0.01 μM and quercetin at a concentration of 0.1; 0.5; 3; 5 mM results in the intensification of glutathione peroxidase. Quercetin at a concentration of 0.1 and 0.3 mM does not change the activity of glutathione-S-transferase, while the studied flavonoid (at a concentration of 0.5; 1; 3; 5 mM) causes a significant increase in the activity of the studied enzyme. Histamine in a concentration of 0.01; 0.1; 1 and 10 μM significantly dose-dependently activate glutathione-S-transferase. The combined action of histamine, at a concentration of 10 µM, and quercetin, at a concentration of 0.1; 0.5; 3; 5 mM, leads to a decrease in enzyme activity in rat erythrocyte hemolysates compared to samples to which only histamine was added, but glutathione-S-transferase values ​​did not reach control limits. Compared to the control, the activity of glutathione-S-transferase increases under the simultaneous action of histamine and quercetin. Simultaneous exposure to histamine at a concentration of 0.01 µM and quercetin causes a significant increase in the activity of glutathione-S-transferase. Histamine has a significant effect on catalase activity in rat erythrocytes, while the combined administration of quercetin and histamine into the blood has a powerful effect on the work of glutathione peroxidase and glutathione-S-transferase.