The purpose of this work is to determine the level of brain neurotrophic factor (BDNF, brain-derived neurotrophic factor) in the blood serum of individuals with alcohol dependence syndrome in the dynamics of alcohol withdrawal syndrome (COA), as well as to compare the results obtained with the clinical picture and the effectiveness of the therapeutic program. Material and methods. The level of BDNF in the blood serum was determined using ELISA. The dynamics of the severity of alcohol withdrawal syndrome (COA), as well as indicators characterizing the psychoemotional status and cognitive abilities of patients, were evaluated using appropriate psychometric scales. The studies were conducted on the 2nd, 8th and 21st days of abstinence. There were examined 70 patients with alcohol dependence syndrome in the dynamics of COA, who met the inclusion criteria and gave informed voluntary consent to participate in the study. Results and discussion. As the symptoms of COA decreased, there was a significant increase in the level of BDNF in the blood serum: on the 21st day of abstinence, the level of BDNF was significantly higher than the values on the 2nd and 8th day of abstinence. On the 8th day of abstinence, BDNF in the blood serum forms a direct correlation with the level of personal anxiety measured using the H scale Ch. Spielberger in the modification of Y.L. Khanin. According to multiple regression analysis on the 21st day of abstinence, BDNF can predict the severity of attraction to alcohol, established using the Pennsylvania scale of attraction to alcohol. Conclusion. Thus, the content of BDNF in the blood serum can act as an independent prognostic marker of the success of care in the treatment of COA.

To study the interrelations between the rs4073 polymorphism of the IL-8 gene, IL-8 serum levels and severity of alcoholic hepatitis (AH).

Some brain-derived neurotrophic factor (BDNF)-targeted miRNAs such as miR-30a-5p may be associated with alcohol addiction; however, their relationship with alcohol withdrawal syndrome (AWS) is not described. We aimed to measure serum BDNF concentration and relative content of miR-30a-5p over the course of alcohol abstinence and compare results with the clinics of AWS. Apart from that, we studied the serum relative content of miR-122, a miRNA that does not target BDNF but is associated with alcohol use disorder. Serum BDNF concentration increases over the course of alcohol abstinence. In contrast, the relative content of miR-122, but not miR-30a-5p, decreased. Moreover, miR-122 (but not miR-30a-5p) negatively correlated with serum BDNF concentrations in the course of AWS. The relative content of miR-122 negatively correlated with depression and anxiety levels on day 8 of abstinence. According to multiple regression analysis, the severity of craving for alcohol and cognitive disturbances may be predictors of serum BDNF concentration on day 21 of abstinence, and vice versa. Thus, serum BDNF concentration and relative content of miR-122 are associated with some aspects of AWS clinical manifestations and may dynamically reflect AWS severity.

Формирование сердечно-сосудистой патологии при чрезмерном употреблении алкоголя сопряжено с повышением концентрации в крови таких медиаторов воспаления, как интерлейкины 6 (IL6) и 8 (IL8) и хемоаттрактанта моноцитов CCL2 (C-C motif ligand 2), а также молекул, участвующих в функционировании эндотелия, в частности фактора роста сосудистого эндотелия (VEGFA, vascular endothelial growth factor А), молекулы клеточной адгезии (ICAM1, intercellular adhesion molecule 1) и эндотелина (EDN1). Предполагается, что данный процесс генетически детерминирован, однако до настоящего момента исследований в этом направлении не проводилось. Целью работы явилось изучение ассоциации носительства аллелей полиморфных локусов, расположенных в генах IL6 (rs1800795), IL8 (rs4073), CCL2 (rs1024611), VEGFA (rs699947 и rs2010963), ICAM1 (rs281437) и EDN1 (rs1800541) с содержанием соответствующих полипептидов в циркуляторном русле и развитием сердечно-сосудистых заболеваний на фоне хронического злоупотребления алкоголем. В исследование были включены лица, злоупотребляющие алкоголем, без выраженной соматической патологии, а также пациенты, у которых на фоне злоупотребления развились заболевания сердечно-сосудистой системы. Уровень IL6, IL8, CCL2, VEGFA, ICAM1 и EDN1 в сыворотке крови оценивали посредством ИФА. Аллели полиморфных локусов были определены посредством ПЦР в режиме реального времени. Установлено, что среди лиц, злоупотребляющих алкоголем, с клинически выраженной патологией сердечно-сосудистой системы значительно чаще встречаются только носители гомозиготного генотипа GG или аллеля G полиморфного локуса в гене IL6 (rs1800795). Кроме того, носительство генотипа GG повышает вероятность развития сердечно-сосудистых заболеваний при хроническом злоупотреблении алкоголем. Однако, дополнительное влияние оказывают демографические факторы и клинические характеристики пациентов. В частности, введение поправки на возраст и пол, а также учет наличия цирроза печени, гипертензии и сахарного диабета, сопровождающих злоупотребление алкоголем, нивелируют повышение риска патологии сердечно-сосудистой системы. Ассоциации полиморфных вариантов в генах IL6 (rs1800795), IL8 (rs4073), CCL2 (rs1024611), VEGFA (rs699947 и rs2010963), ICAM1 (rs281437) и EDN1 (rs1800541) с содержанием белковых продуктов соответствующих генов в циркуляторном русле выявлено не было. Cardiovascular diseases in alcohol abusers are associated with elevation of plasma levels of proinflammatory cytokines such as IL6, IL8 and CCL2 as well as molecules involved in endothelial functioning including VEGFA, ICAM1 and EDN1. This phenomenon is supposed to be genetically determined. However to date the issue has not been investigated. Thus, we aimed to study the relationship between carriage of SNPs of IL6 (rs1800795), IL8 (rs4073), CCL2 (rs1024611), VEGFA (rs699947 and rs2010963), ICAM1 (rs281437) and EDN1 (rs1800541) genes with the serum levels of their products and the development of cardiovascular diseases in alcohol abusers. The study included alcohol abusers without apparent somatic pathology and alcohol abusers with clinical manifestations of cardiovascular disease. Serum levels of IL6, IL8, CCL2, VEGFA, ICAM1 and EDN1 were estimated by EIA. SNPs were determined by means of real-time PCR. We found that among the SNPs studied only carriers of homozygous GG genotype and G allele of IL6 (rs1800795) were more frequent in alcohol abusers with cardiovascular diseases. Moreover, carriage of homozygous GG genotype of IL6 (rs1800795) increases the probability of development of cardiovascular pathology in alcohol abusers. However, adjustment for age, gender and the presence of liver cirrhosis, hypertension and diabetes mellitus as co-variates eliminates the enhanced risk of cardiovascular pathology. Polymorphisms of IL6 (rs1800795), IL8 (rs4073), CCL2 (rs1024611), VEGFA (rs699947 and rs2010963), ICAM1 (rs281437) and EDN1 (rs1800541) did not determine serum levels of the related polypeptides.

Фиброз печени и его конечная стадия - цирроз являются одной из причин летальных исходов при хронических заболеваниях печени. Алкоголь и его метаболиты вызывают накопление внеклеточного коллагена, приводящего к фиброзу, вследствие цитокинового дисбаланса, процессов фиброгенеза и фибролиза. Цель. Изучить иммуновоспалительные факторы, ассоциации полиморфизма гена коллагена I типа COL1A1_1 C/A (rs1107946) с развитием фиброза печени у пациентов, злоупотребляющих алкоголем. Методика. Генотипировано 46 пациентов с фиброзом печени, злоупотребляющих алкоголем (срок употребления алкоголя 15,6±9,5 лет). Группы сформированы в зависимости от стадии фиброза печени, определенного методом непрямой эластометрии: 1-я - F0+1 (n=20), 2-я - F3+4 (n=36). Критерии исключения: острый алкогольный гепатит, хронические заболевания печени неалкогольного генеза, общие воспалительные и аутоиммунные процессы. У всех больных определяли концентрации интерлейкина (ИЛ)-6, ИЛ-8, ФНО-альфа, VEGF-A, s-ICAM-1, ЭТ-1. Полиморфизм гена COL1A1_1 определяли методом Real-Time PCR. В качестве контрольной группы полиморфизма гена коллагена были обследованы 30 здоровых мужчин и женщин. Результаты. Уровни ИЛ-6, ИЛ-8, s-ICAM-1, ET-1 зависили от степени алкогольного фиброза печени: во 2-й группе они были выше, чем в 1-й (p<0,05). Плотность печени статически значимо (p<0,05) коррелировала с уровнем ИЛ-6 (r=0,6), ИЛ-8 (r=0,77), s-ICAM-1 (r=0,58), ET-1 (r=0,56). Cтатически значимых различий уровней ФНО-альфа и VEGF-A между группами не выявлено (р>0,05). У больных с фиброзом печени, страдающих алкогольной зависимостью, частота встречаемости аллеля А статически значимо выше, чем в контрольной группе (38,3% против 12,5%, χ2 = 7,6212; р=0,005768). Частота гетерозигот СА существенно не отличается в группе больных и контрольной группе (38% и 25% соответственно, р=0,4425). Гомозиготы АА были выявлены только в группе больных с фиброзом печени, частота встречаемости составила 19,1%. Заключение. Выявлена прямая корреляционная связь между уровнем цитокинов (IL-8, IL-6), молекул эндотелиальной дисфункции (EТ1, sICAM-1) с уровнем фиброза печени. Наличие аллеля А гена COL1A1_1 может являться одним из генетических факторов развития фиброза печени у больных, злоупотребляющих алкоголем. Liver fibrosis and its final stage - cirrhosis is one of the causes of deaths in chronic liver diseases. Alcohol and its metabolites mediate the accumulation of extracellular collagen, leading to fibrosis mediated by cytokine imbalances, processes of fibrogenesis and fibrolysis. The purpose. To investigate the immune-inflammatory factors and association between the COL1 A1_1 C/A (rs1107946) polymorphism and liver fibrosis in patients abusing alcohol. Мethods. 46 patients were genotyped (the period of alcohol consumption was 15.6±9.5 years) with liver fibrosis, alcohol abusers. They were divided into 2 groups depending on the stage of liver fibrosis: 1- F0+1, n=10; 2- F3+4, n=36. Exclusion criteria were chronic non-alcoholic liver disease, acute alcoholic hepatitis, general inflammatory and autoimmune diseases. We analyzed serum concentrations of interleukin (IL)-6, IL-8, TNF-α, VEGF-A, s-ICAM-1, ET-1 and polymorphism of the COL1A1_1 gene was determined using the Real-Time PCR analysis. The stage of liver fibrosis was identified with the elastography method (FibroScan, Echosens, France). Results. The levels of IL-6, IL-8, s-ICAM-1, ET-1 depended on the degree of alcoholic liver fibrosis. The liver fibrosis was significantly correlated with the level of IL-6 (r = 0.6), IL-8 (r = 0.77), s-ICAM-1 (r = 0.58), ET- 1 (r = 0.56) (p <0.05). There were no significant differences in the levels of TNF-alpha and VEGF-A between two groups (p> 0.05). It was found that the frequency of allele A among patients was 38.3% which is significantly higher than in the control group (12.5%; р=0.005768). The frequency of heterozygotes CA was not significantly different between the group of patients and the control group (38% and 25% respectively, p = 0.4425). Homozygous AA genotypes were revealed only in patients with liver fibrosis, the frequency was 19.1%. Conclusion. A positive correlation between the level of cytokines (IL-8, IL-6), endothelial dysfunction molecules (ET1, sICAM-1) and the level of liver fibrosis was revealed. The presence of allele A of the COL1A1_1 gene may be one of the genetic factors involved in the development of liver fibrosis in patients abusing alcohol.

Background: Uncontrolled use of alcohol can lead to the development of cirrhosis of the liver, which is manifested by fibrosis with the formation of regenerative nodes, an increase in pressure in the portal vein system and impaired liver function. Hepatic endothelium dysfunction during the formation of portal hypertension is accompanied by an increase in the level of protein molecules involved in the functioning of the endothelium: vascular endothelial growth factor A (VEGF-A), a soluble form of the intercellular adhesion molecule (s-ICAM-1) and endothelin-1 (ET -one). It is assumed that elevated levels of VEGF-A, s-ICAM-1 and ET-1 in alcoholic liver cirrhosis (AHC) may be interconnected with the structure of polymorphic loci, the promoter regions of the respective genes, which in turn may be a genetic risk factor for developing cirrhosis.Aims: Investigate the relationship of carriage of variant forms of polymorphic loci located in the promoter regions of VEGF-A, ICAM-1 and ET-1 with the level of the corresponding proteins in the blood serum and the risk of AHC.Materials and methods: The main group consisted of patients with pathological dependence on alcohol, aggravated by cirrhosis of the liver (AHC, n=60). The control group consisted of persons suffering from alcohol abuse, without liver pathology (AA, n=24). The observation period was the period of hospitalization. The serum levels of VEGF-A, s-ICAM-1 and ET-1 were evaluated by enzyme immunoassay. The distribution of variant forms of polymorphic loci located in the promoter regions of the VEGF-A genes (rs699947 and rs2010963), ICAM1 (rs281437) and ET-1 (rs1800541) in the studied sample was performed by real-time PCR.Results: The development of alcoholic cirrhosis was accompanied by a significant increase in the concentration of VEGF-A, s-ICAM-1 and ET-1 in serum. At the same time, direct correlations between the concentrations of VEGF-A, s-ICAM-1 and ET-1 in serum and the diameter of the portal vein in persons with liver cirrhosis were revealed. Patients with AHC are often carriers of the G allele of rs1800541 locus, located in the promoter of the ET-1 gene, compared with individuals suffering from control without liver pathology, which is associated with an increased risk of developing cirrhosis in alcohol dependence. The carriage of the C allele rs699947, as well as the C allele rs2010963 located in the promoter of the VEGF gene was associated with an increased level of VEGF-A in the AHC compared to carriers of this allele in the AA group. In addition, in the group of patients with AHC, carriers of allele C, homozygous CC genotype and heterozygous GC genotype of rs2010963 locus compared with carriers of G allele or homozygous GG genotype, respectively, were characterized by elevated serum VEGF-A levels.Conclusion: Carrier allele G of the rs1800541 locus (ET-1) is a risk factor for liver cirrhosis with alcohol abuse. The carriage of the C allele rs699947, as well as the C allele rs2010963 located in the promoter of the VEGF gene, can determine the elevated serum VEGF-A level in the AHC.

КЛИНИЧЕСКАЯ ФАРМАКОЛОГИЯ И ТЕРАПИЯ, 2018, 27 (5) Значение иммуновоспалительных и генетических факторов в развитии алкогольного фиброза печениА.С.Иванов 1 , И.В.Гармаш 1 , О.С.Аришева 1 , Н.Н.Теребилина 2 , В.Ю.Баронец 2 , Д.И

Aim. To estimate the contribution of liver cirrhosis (LC) to the development of heart diseases in alcohol abusers. Subjects and methods. The investigation included 80 patients with alcoholic LC without a history of cardiovascular and respiratory diseases and, as a control group, 32 alcohol abusers without a history of chronic diseases of the liver and cardiovascular and respiratory systems; 45 patients with alcoholic cardiomyopathy (ACM) and congestive heart failure without a history of coronary heart disease and valvular diseases, among whom 11 patients were found to have LC. In addition to standard clinical examination, all the patients underwent electrocardiography, by estimating the corrected QT interval (QTc), standard echocardiography; and those without ACM underwent estimation of left ventricular (LV) kinetics using speckle-tracking echocardiography. Results. The patients with alcoholic LC were found to have a higher LV ejection fraction and a more obvious impairment of LV global longitudinal deformity, and more commonly LV diastolic dysfunction. 16 of the 80 patients with LC were observed to have moderate pulmonary hypertension while the mean pulmonary artery pressure (MPAP) was within the normal range in all the patients without LC. A prolonged QTc interval was revealed in the patients with LC. The duration of QTc was directly correlated with the MELD severity of LC. The patients with chronic heart failure in the presence of ACM and CL showed a more obvious LV diastolic dysfunction, as estimated by E/E’, a greater LV mass index, and a higher MPAP than those with ACM without LC. Conclusion. The LC patients both with ACM and without a history of diseases of the heart were noted to have its more evident disorders as diastolic dysfunction and elevated MPAP. Those without ACM were observed to have impaired LV global deformity and a prolonged QTc interval.

Parameters reflecting oxidative stress (OS) have been studied in 37 patients with alcoholic liver disease (ALD) during admission to the hospital and 2 weeks after the beginning of therapy. The patients were divided into 3 groups: alcoholic hepatitis (AH), alcoholic cirrhosis with hepatic insufficiency (the group C by the Child-Paquet scale) and terminal stage patients (subsequently died). All patients were characterized by a significant increase in plasma products of lipid peroxidation (conjugated diene and malondialdehyde) and a decrease of the ceruloplasmin level. The coefficient C OS significantly exceeded normal values both on admission and after the 2-week course of traditional therapy. This suggests an important role of the OS with ALD.

AIM:To estimate the contribution of immuno-inflammatory changes to the formation of clinical and hemodynamic features in alcoholic patients with chronic heart failure (CHF).SUBJECTS AND METHODS:Forty-five males with CHF in the presence of alcohol-induced heart damage (AIHD) who had been admitted to therapeutic units for decompensated heart failure were examined. A control group consisted of 20 men with the CHF severity comparable with the NYHA classification in the presence of prior myocardial infarction. All the patients underwent examination of the immune-inflammatory status--the cytokines: interleukin (IL) 6, IL-8, IL-12, tumor necrosis factor-alpha (TNF-alpha), transforming growth factor-beta1, endotoxin, cellular immune parameters, and cardiac structure and function by echocardiography.RESULTS:The patients with CHF in the presence of AIHD, as compared to those with ischemic cardiomyopathy, showed the higher levels of inflammatory cytokines (IL-6, TNF-alpha, IL-12, and endotoxin) and cell-mediated immunity changes (the smaller count of suppressor T cells, natural killer cells, and a shift of the T-helper/T-suppressor ratio towards the T-helper population). The magnitude of these changes correlated with the severity of CHF and cardiac morphofunctional changes.CONCLUSION:The relationship of immuno-inflammatory changes to the severity of CHF and the morphofunctional state of the heart irrespective of the etiology of heart failure demonstrated the role of immune inflammation in its pathogenesis particularly in alcoholic patients who were found to have more marked immuno-inflammatory changes than in those with ischemic cardiomyopathy.

We had analyzed the diagnostic findings of 42 cirrhosis patients with infectious complications in the research. There have been evaluated the diagnostic significance of inflammatory markers determination (endotoxin, D-dimers, C-reactive protein) under out-of-hospital and nosocomial infections and its dynamics during antibacterial therapy.

Glutamatergic neurotransmission has been suggested to modulate cue-induced drug-seeking behavior. Here we examined the effects of metabotropic glutamate receptor agonists on alcohol self-administration and cue-induced reinstatement. Rats were trained to self-administer 10% w/v ethanol under an FR1 schedule of reinforcement during 30-min sessions. In the reinstatement experiments, ethanol and a non-rewarding quinine solution (available on alternating days) were paired with olfactory stimuli (S+/S−) as well as light (CS+) or tone (CS−) stimuli. Following extinction training, reinstatement of responding was induced by the ethanol-associated stimuli (S+/CS+). The mGlu2/3 receptor agonist LY379268 (0, 1, 3 and 5 mg/kg i.p.) and the mGlu8 receptor agonist (S)-3,4-DCPG (0, 5, 10 and 15 mg/kg i.p.) attenuated alcohol self-administration and reinstatement at doses that decreased also spontaneous locomotor activity. The results suggest that metabotropic glutamate receptors may have a role in the modulation of alcohol seeking and self-administration. However, further studies with ligands with fewer motor-suppressant side effects are needed.

We studied the effect of bioactive peptide HLDF-6 on functional activity of the endogenous antinociceptive system in the offspring of morphine-tolerant animals. Disturbances in this system included changes in the thermonociceptive threshold and enkephalinase A activity in various brain structures. The peptide acted as a potent regulator of the homeostasis in systems responsible for the synthesis and catabolism of endogenous opioids. HLDF-6 effectively corrected disorders of the endogenous antinociceptive system.

We carried out a complex physiological, neurochemical, and neuroimmunologic study of the formation of tolerance to analgetic effect of morphine and analyzed enkephalinase A activity in different brain structures and serotonin antibodies in the serum. More early development of morphine tolerance and a sharp increase in serum antibody titer was found in the offspring of morphine-tolerant rats. This points to an imbalance in the neurotransmitter system and can serve as a diagnostic marker of endogenous opioid system pathology.

RATIONALE:Regulatory neuropeptide systems appear to modulate anxiety and emotionality, since anxiety in rats can be increased by intracerebroventricular (ICV) administration of diazepam-binding-inhibitor fragment (DBI) and decreased by ICV administration of neuropeptide Y (NPY) or substance P (SP).OBJECTIVE:Involvement of these three neuropeptides in genetic predisposition to anxiety was studied in two inbred rat strains.METHODS:Levels of anxiety to novel environments were first measured in Fischer-344 (F-344/N) and Wistar Albino Glaxo (WAG/G) rats using open-field conflict, hole-board, black and white box, elevated-plus maze and Vogel lick suppression procedures. Levels of SP, DBI and NPY in the hippocampus, midbrain and hypothalamus of F-344/N and WAG/G rats were then measured without stress (basal levels) or after stress induced by shuttle-box, shock-avoidance testing. Finally, effects of ICV injection of SP or NPY rats were measured in F-344/N and WAG/G rats using the hole-board test.RESULTS:F-344/N rats had elevated level of anxiety compare to WAG/G rats with all five procedures. Levels of SP in the hippocampus, midbrain and hypothalamus of F-344/N rats were significantly lower than in WAG/G rats and levels of SP decreased in WAG/G, but not F-344/N, rats after stress. Levels of DBI in the hippocampus and midbrain of F-344/N rats were also lower than in WAG/G rats, but they increased in F-344/N rats after stress. In contrast, levels of NPY were higher in the midbrain of F-344/N rats than in WAG/G rats, especially after stress. ICV injection of SP or NPY decreased anxiety in the black and white box in both F-344/N and WAG/G rats, but F-344/N rats were more sensitive.CONCLUSIONS:These findings support the hypothesis that decreased levels of SP in certain brain regions may contribute to high levels of anxiety in rats. Decreased levels of DBI and increased levels of NPY in high-anxiety animals may act as compensatory mechanisms.

The use of naloxone hydrochloride (0.2-0.4 mg) in complex therapy of adolescent heroin addicts significantly prolonged the half-life of serum leu-enkephalin, slightly elevated the thresholds of thermal nociceptive reactions, and improved some clinical indices (considerably reduced drug addiction, eliminated affective disorders, etc.), which are important for deactualization of drug addiction and promoting remission.