Content of sP-selectin and Cytokines in Blood of Patients with Type 2 Diabetes Mellitus and Arterial Hypertension Depending on Diabetes Compensation Condition

The goal of our research has been to study sР-selectin and cytokine content changes, namely ІL-2, ІL-6 and TNF-α in blood plasma of patients with type 2 diabetes mellitus with varying compensation for the disease and arterial hypertension, as well as to investigate a possible interrelation between sР-selectin and cytokines. To achieve the goal 137 patients with type 2 diabetes mellitus with AH of the І-ІІ stages and without AH (72 women and 65 men) have been examined. The levels of sР-selectin, ІL-2, ІL-6, TNF-α in blood serum were determined by means of immunoenzymatic assay method. As a result, a reliable increase in sP-selectin level in blood serum has been detected in patients with type 2 diabetes mellitus with deterioration of diabetes compensation along with АH; increase in ІL-6 level in groups with good and satisfactory compensation for diabetes and TNF -α in groups with bad compensation with the presence of АH. A reverse correlation between the content of sP-selectin and TNF-α in groups of those examined with non-compensated type 2 DM, associated with АH and without AH, stronger in a group of patients with type 2 DM and AH. The level of sP-selectin in blood increases with the deterioration of type 2 DM compensation along with AH. Most probable reverse correlations between sP-selectin and TNF-α levels in groups of patients with insufficient compensation for diabetes may indicate a mutually potential role of these factors in the development and progression of type 2 DM decompensation and AH.


Introduction
Diabetes mellitus (DM) and arterial hypertension (АH) are among the most common diseases in the world.Over 80% of patients with type 2 diabetes mellitus suffer from arterial hypertension.Especially dangerous is a combination of these diseases, as long as in this case cardiovascular risk and death rate in this category of patients significantly increases.P-selectins are adhesive molecules, which are exposed with erythrocytes and platelets in the process of activation, they provide their interaction with leukocytes.Р-selectin of erythrocytes is kept in Weibel-Palade bodies.Its production is stimulated by inflammatory response cytokines, hyperglycemia and vasospasm.Р-selectin plays an important role in the process of endothelial cell interactions with blood cells, coagulation, blood clot formation, including atheromatous plaques and is considered to be the marker of hyperactivity of blood clots.A soluble sР-selectin is detected in the blood Violation of Р-selectin expression may be one of the factors in development of cardiovascular events (Aref S. et al., 2005;Glowinska B. et al., 2005;Jiao J.A. et al., 2010).Up to now discussing and less researched is the problem of sР-selectin in the pathogenesis of endothelial dysfunction development in patients with diabetes and its taking part in acute cardiovascular conditions.R. Füth and other authors has revealed direct connection between diastolic dysfunction of myocardium and the level of sР-selectin despite existence of diabetes and ischemic heart disease.However, such scientists as V. Bláha, T Dogru and К. Gokulakrishnan in their research have discovered reliable increase of sР-selectin in patients with type 2 diabetes, which closely correlated with weight index, markers HOMA R and hyperglycemia.(Bláha V et al., 2006;Dogru T et al., 2006Gokulakrishnan K et al., 2006).In case of АH and type 2 DM combination, a complex of not only hemodynamic changes, impairment of blood coagulation properties, specific to HD, is observed, but also disorders of carbohydrate and lipid metabolisms and signs of systemic inflammation, peculiar to type 2 DM are notable (Yang HI. et al., 2016).Data analysis in contemporary literature proves that chronic systemic inflammation is an important component of type 2 DM development, atherogenesis, remodeling of vascular wall, development of AH, IHD.Cytokines are the endogenous mediators, which regulate the intensity and duration of an immune inflammatory response.Normally, they realize interrelation between non-specific resistance of the body and specific defense.A numerous group of cytokines, especially interleukin (ІL) and tumor necrosis factor α (TNF-α), participate in regulation of the basic body functions both in normal functioning and in a pathological condition.They provide close correlation between different groups of cells (Mattu H. S. et al., 2013).It is supposed that type 2 DM is a result of acute-phase inflammatory response activations, in the course of which, release of cytokines occurs (Corbin KL. et al., 2016;Ibarra Urizar A. et al., 2016).However, at present the biological and pathophysiological effects of P-selectin in the human body, moreover, the influence of P-selectin on diabetes compensation condition and the course of AT, its interrelations with other adipocytokines have been investigated partially and this problem remains the theme of scientific discussions.Thus, the goal ofour investigation was to study the changes of sР-selectin and cytokine contents, namely ІL-2, ІL-6 and TNF-а, in blood plasma of patients with type 2 DM with varying compensation for the disease and arterial hypertension, as well as to investigate a possible interrelation between sР-selectin and cytokines.

Materials and Methods
The investigation involved 137 patients with type 2 DM with АH of the І-ІІ stages and without АH (72 women and 65 men), who sought consultation at the endocrinology department of Danylo Halytskyi Lviv National Medical University.Anthropological examinations, which included measurement of the patients' weight and height, were conducted for all patients.Presence of obesity and its degree was diagnosed based on classification criteria of the World Health Organization (1997) with determination of body mass index (BMI): according to the formula: BМI = body weight (kg) / height (m 2 ).The diagnosis of type 2 DM was made according to the WHO experts' recommendations.Compensation for type 2 DM was verified according to the order of the Ministry of Health of Ukraine No 1118 dated 21.12.2012.The diagnosis of АH was made based on case history data, clinical and instrumental examination after exclusion of symptomatic hypertension according to WHO recommendations.
After standard procedure of diagnostics, the patients' daily blood pressure was monitored with electronic automatic device for measurement of blood pressure VAT41-2.Analysis of data was carried out by means of «АRIАDА» software (with automatic calculation of standard indices).
Statistical processing of the results was performed by means of parametric methods of Student t-test and correlation analysis with determination of Pearson's code, and in case of normal distribution inconsistency-by means of Kolmogorov-Smirnov test and correlation analysis with determination of Spearman's correlation coefficient.While comparing the groups, ANOVA test was applied.The results are presented in the form of the mean value and standard deviation and correlation coefficient r.The reliable value was considered to be the one at Р<0.05.

Differences in compensation
The level of s with the group found that in increases sign ТNF-α level i group of patie 2 DM compen   and АH.ІL-2, in its turn, activates cytotoxic T-cells, monocytes, microphages, which increase the synthesis and secretion of ІL-6, ТNF-α.The latter exhibits a similar activity as ІL-2 concerning inflammatory and immune reactions (Nelaeva AA. et al., 2013).
AH is associated with endothelial dysfunction and oxidative stress.Endothelial dysfunction is considered to be an important component of insulin resistance in type 2 diabetes mellitus and АH.It results in activated state, characterized by increased agglutination and platelet aggregation, hypersecretion of Р-selectin by platelet membranes.It was also observed in our patients in case of compensation deterioration for type 2 DM with AH.It has been proven that Р-selectin level is increased in type 2 diabetes mellitus with marked angiopathies (Blankenberg S. 2003;Penman A. et al., 2015;Kolahdouz P. et al., 2015).Respectively, the increased level of sP-selectin may be regarded as an early marker of vascular complications of type 2 DM (Woollard KJ. et al., 2014).D. Bednarska-Chabowska with co-authors, investigating the level of selectin in patients with types 1 and 2 diabetes mellitus, revealed that in case of type 2 DM , associated with AH, presence of AH causes increase in P-selectin level, which coincides with our results.Moreover, these authors made a conclusion that the degree of endothelial tissue damage in diabetes mellitus depends on concomitant hypertension.P-selectin concentration increase is proportional to the degree of existing damage to vascular wall (Bednarska-Chabowska D. et al., 2002).

Conclusions
The level of sP-selectin in blood increases with deterioration of type 2 diabetes mellitus compensation and АH.
Highly reliable correlations between the levels of sP-selectin and ТNF-α in groups of patients with poor compensation for diabetes mellitus may confirm a mutually potential role of these factors in the development and progression of type 2 DM decompensation and AH.

Figure
Figure 1.Co

Table 3 .
Corre DM and AH a