Abstract

The aim of the work was to study the parameters of hemostasis and their pathogenetic relationship with pro-inflammatory cytokines in hematogenous thrombophilia. Materials and methods: to solve the problem, a prospective study was conducted for the period 2021-2022. The main group included 47 patients registered with the RSSPMC of hematology with diagnoses of chronic leukemia included in the group. The selected patients were aged 18 to 45 years with a history of thrombosis and 36 patients with thrombosis at the moment. The comparison group consisted of 30 practically healthy people. Results and conclusions: At present, in patients with thrombosis, the intensity of the synthesis of positive factors IL-6 and IL-8 prevails over the synthesis of other cytokines, while in the synthesis of negative cytokines TNFα significantly prevails, which apparently determines the severity of thrombosis. A pathogenetic relationship between increased IL-1β, TNF-α, and activation of procoagulant mechanisms in the blood of patients suffering from acute thrombosis was revealed.

1. Introduction

Currently, the course of hematogenous thrombophilia, the severity and prevalence of thrombotic complications are quite unpredictable, thrombosis has become increasingly common in young people. According to current data, thrombophilia is the third leading cause of vascular death ^{1, 5}. Incidence rates increase from 1 per 10,000 per year among people under 40 years old, more than a third of cases occur in people over 60 years old, almost 1% per year occurs in people over 80 years old ^{5, 7}.

Since the etiology of thrombosis is multifactorial, the presence of a thrombophilic defect is only one of many elements that determine this risk ^{3, 4}. In this regard, the study of the synthesis of inflammatory cytokines produced by cells of the immune system and affecting hematopoiesis (being its positive and negative regulators), in normal conditions and in thrombophilias, seems relevant in studying the role of the cytokine system in the formation of a thrombus and the tendency to thrombosis.

The aim of the work was to study the parameters of hemostasis and their pathogenetic relationship with pro-inflammatory cytokines in hematogenous thrombophilia.

2. Materials and Methods

To solve this problem, a prospective study was conducted for the period 2021-2022. The main group included 47 patients registered with the RSSPMC of hematology with diagnoses of chronic leukemia included in the group. The selected patients were aged 18 to 45 years with a history of thrombosis and 36 patients with thrombosis at the moment. The comparison group consisted of 30 practically healthy people comparable in age. The criteria for inclusion in the main study group were the results of clinical, laboratory-instrumental and ultrasound examinations, characteristic of thrombophilia. Exclusion criteria from the study were pregnancy, endocrine diseases, malignant neoplasms, pulmonary and heart failure in patients.

The assessment of indicators of the state of coagulation hemostasis, fibrinolysis, cytokine status in patients of the main group was carried out at the time of their admission to the department, before the start of treatment against the background of severe clinical manifestations of pathology.

Results and its discussion. From the works of domestic and foreign authors, it is known that pro-inflammatory cytokines have the ability to change the adhesive-aggregation properties of platelets, erythrocytes, to initiate the expression of endothelial factors involved in the regulation of blood coagulation and fibrinolysis activity ^{3, 6}.

Thus, the most important regulator of vascular-platelet hemostasis, blood coagulation and fibrinolysis is IL-1β, which increases the content of aggregation stimulators, procoagulants, fibrinolysis inhibitors, in particular von Willebrand factor and platelet activation factor ^{7, 8}. IL-1β and TNF-α activate the blood coagulation process, stimulating the expression of tissue factor on endotheliocytes and monocytes, preventing the formation of thrombomodulin, and reducing the activating effects on protein C ¹⁰.

In this regard, it was advisable to find out whether pro-inflammatory cytokines play the role of mediators that ensure the implementation of procoagulant effects in acute thrombosis. To partially address this issue, the content of IL-1β, IL 4, IL 6, IL 10, TNF-α and INFγ in the peripheral blood during the period of pronounced clinical manifestations was studied.

Spontaneous production of IL-1β in the peripheral blood of patients (Table 1), with thrombosis at the moment (group 2) and a history of thrombosis (group 3) compared with the norm (group 1), was increased in groups 2 and 3 of patients (7.5±1.1 pg/ml), but according to the average statistical values it was not significant (p>0.05), however, when compared with the donor group (1.3±0.2), a significant difference was revealed p<0.001.

The level of spontaneous production of IL-6 was increased by 26 times in patients of group 2, and by 17 times in patients with a history of thrombosis compared with the norm (63.2±5.4 pg/ml, 42.11±9.49 pg /ml and 2.4±0.2 pg/ml, respectively). And the spontaneous production of IL-10 in the peripheral blood was 5 times higher than that in donors 15.1±2.8 pg/ml and 2 times in patients with a history of thrombosis 7.3±1.1 pg/ml, in contrast from a group of healthy donors 3.61±0.64 pg/ml (correspondingly, P1-2<0.001; P2-3>0.05).

Thus, the indicators of spontaneous production of IL-1β, IL-6 and IL-10, which in patients with acute thrombosis and a history of thrombosis (3 months) were increased by more than 10 times, apparently indicate that the process thrombus formation directly depends on the level of pro-inflammatory cytokines and their concentration in the peripheral blood, and as a result of the hypercoagulable state.

TNFα and IFNγ have been studied as negative regulators of hematopoiesis. Spontaneous synthesis of IFNγ in peripheral blood was almost 5 times higher than that in the donor group (16.2±1.1 pg/ml, 12.45±1.18 and 3.33±0.19 pg/ml, respectively). Another negative hematopoietic factor (TNFα), during acute thrombosis, as we found, sharply, 660 times higher than the average value in donors, and in patients with a history of thrombosis, it was 550 times higher than the conventionally accepted normative value (13.2±3.2 pg/ml, 11.02±4.6 and 0.02±0.003 pg/ml, respectively) (Table 1).

Thus, it can be considered that the negative regulators of hematopoiesis (TNFα and IFNγ) have a significant effect on the spontaneous production of pro-inflammatory cytokines in patients with current thrombosis and a history of thrombosis.

Spontaneous production of IL-4 in peripheral blood was comparable to the norm (>0.05). IL-4 is a direct TNFα antagonist, and a strong inverse correlation was found between these two cytokines (r = -0.63). Therefore, in general, taking into account the hyperproduction of TNFα, characteristic of thrombosis, it can be said that the cells of the immune system in this disease synthesize an insufficient amount of the antagonist of negative factors IL-4.

Comparing the levels of spontaneous production of cytokines in the group of donors and in the group of patients, we can conclude that normally, the intensity of the production of positive factors IL-6, TNFα and IFNγ prevails over the synthesis of other cytokines in the peripheral blood.

3. Conclusions

1. In various diseases accompanied by intravascular coagulation against the background of leukemia, changes in the composition of cytokines are observed. The level of TNF, IFN and IL6 and the production of positive hematopoietic factors by peripheral blood mononuclear cells: IL-1β, and IL-8 in chronic myeloproliferative diseases are tenfold increased and have pathogenetic significance in the development of hematogenous thrombophilia.

2. The development of thrombosis in these pathologies is due to the fact that peripheral blood mononuclear cells produce an ultra-high amount of the cytokine TNFα, which inhibits hematopoiesis, and a high amount of IFNγ.

3. Cells of the immune system in CMPD synthesize an insufficient amount of the cytokine IL-4, which is an antagonist of the cytokine TNFα, which inhibits hematopoiesis.

4. In patients with thrombosis, at the moment, the intensity of the synthesis of positive factors IL-6 and IL-8 prevails over the synthesis of other cytokines, while in the synthesis of negative cytokines, TNFα significantly prevails, which apparently determines the severity of thrombosis.

5. A pathogenetic relationship between increased IL-1β, TNF-α, and activation of procoagulant mechanisms in the blood of patients suffering from acute thrombosis has been revealed.

6. Determination of the levels of IL-1β, TNF-α in blood serum in patients with acute thrombosis can be used to predict the development and timely correction of shifts in the coagulation potential of the blood, which will expand the possibility of successfully choosing an adequate treatment strategy and timely prevention of the development of complications of the thrombotic process.

References