Современные методы исследования атеросклероза и ишемической болезни сердца: проточная цитометрия

 The problem of atherosclerosis, which forms the pathological basis of coronary artery disease (CAD), is one of the most discussed ones in development of cardiovascular diseases. This chronic inflammatory disease involves interactions between different cells, and an atherosclerotic plaque is a complex immunological environment. Modern  quantitative methods  increase the understanding of the pathophysiological processes responsible  for progression of atherosclerotic plaques. Flow cytometry is a powerful modern method that allows for a complex and simultaneous cell analysis. This review is devoted to studies on atherosclerosis and CAD performed  using flow cytometry.   Проблема атеросклероза, формирующего патологическую основу ишемической болезни сердца, является одной из наиболее обсуждаемых в развитии сердечно-сосудистых заболеваний. Это хроническое воспалительное заболевание  включает комплекс сложных взаимодействий между различными клетками, а атеросклеротическая бляшка представляет собой сложную иммунологическую  среду. Современные количественные методы повышают  понимание патофизиологических процессов, ответственных  за прогрессирование атеросклеротической бляшки.  Проточная цитометрия представляет собой мощный  современный метод, позволяющий проводить комплексный анализ клеток одновременно. Данный обзор посвящен  научным исследованиям атеросклероза и ишемической  болезни сердца, выполненным с помощью метода проточной цитометрии.

Klotho protein in men with type 2 diabetes mellitus blood and its association with cardiometabolic risk factors

The aim of the study was to investigate Klotho protein levels in men with type 2 diabetes blood and its associations with several cardiometabolic risk factors. Material and methods. The study included 37 men with diabetes and 141 men without diabetes. Fasting blood samples were collected to measure Klotho protein levels and some biochemical parameters. Results and its discussion. The Klotho protein level in men with diabetes was significantly lower than in men without diabetes (374 [117; 500] and 515 [315; 1009] pg/dl, p<0.0001). Among the examined men with diabetes with a glomerular filtration rate of less than 60 ml/min/1.73 cm2, the concentration of Klotho protein was 4 times lower than in the comparison group (104 [93; 118] and 413 [147; 535] pg/dl, p = 0.014) In men with diabetes, the Klotho protein was inversely correlated with the ratio of waist to hip circumference (-0.329; p = 0.047). But with multivariate analysis, only a tendency towards a negative association of the Klotho protein with abdominal obesity was determined (-0.385, p = 0.078). Conclusion. The content of Klotho protein in men with diabetes is significantly lower, especially in middle-aged men and in those with a reduced glomerular filtration rate. In men with diabetes, the Klotho protein has a negative correlation with the presence of abdominal obesity. In a multivariate analysis among men with diabetes, the Klotho protein tends to be inversely associated with the presence of abdominal obesity

Longitudinal trajectories of blood lipid levels in an ageing population sample of Russian Western-Siberian urban population

This study investigated 12-year blood lipid trajectories and whether these trajectories are modified by smoking and lipid lowering treatment in older Russians. To do so, we analysed data on 9,218 Russian West-Siberian Caucasians aged 45-69 years at baseline participating in the international HAPIEE cohort study. Mixed-effect multilevel models were used to estimate individual level lipid trajectories across the baseline and two follow-up examinations (16,445 separate measurements over 12 years). In all age groups, we observed a reduction in serum total cholesterol (TC), LDL-C and non-HDL-C over time even after adjusting for sex, statin treatment, hypertension, diabetes, social factors and mortality (P 60 years at baseline). In smokers, TC, LDL-C, non-HDL-C and TG decreased less markedly than in non-smokers, while HDL-C decreased more rapidly while the LDL-C/HDL-C ratio increased. In subjects treated with lipid-lowering drugs, TC, LDL-C and non-HDL-C decreased more markedly and HDL-C less markedly than in untreated subjects while TG and LDL-C/HDL-C remained stable or increased in treatment naïve subjects. We conclude, that in this ageing population we observed marked changes in blood lipids over a 12 year follow up, with decreasing trajectories of TC, LDL-C and non-HDL-C and mixed trajectories of TG. The findings suggest that monitoring of age-related trajectories in blood lipids may improve prediction of CVD risk beyond single measurements

The frequency of metabolic syndrome and its individual components in women aged 25–45 years, depending on the level of prolactin

Background: Hyperprolactinemia is one of the most common hypothalamic-pituitary-endocrine disorders in women of reproductive age, with the highest frequency at the age of 25–44 years. In addition to influencing the reproductive system, it is important to study the effects of prolactin (PRL) on various metabolic links. Available data indicate that the effect of PRL on metabolism depends on its level. In this regard, the study of the relationship of different levels of PRL with anthropometric parameters, indicators of lipid and carbohydrate metabolism in young women is relevant.Aim: To study the frequency of metabolic syndrome (MS) and its individual components in women aged 25–45 years with different levels of prolactin.Materials and methods: Work design — cross-sectional research. A randompopulationsample of women 25–45 agedwas examined. Pregnant and breastfeeding women with macroprolactinoma, and taking antipsychotics were excluded. Information was collected using a structured ­questionnaire, including, but not limited to, the presence of pregnancies, childbirth, menstrual irregularities, and a clinical examination, anthropometric measurements, biochemical and hormonal blood analyzes were performed. Statistical data processing was carried out.Results: According to the inclusion and exclusion criteria, this analysis presents data from 401 women, the average age of the examibed was 36.14±6.19 years. There was no difference in the levels of thyroid-stimulating hormone and prolactin (PRL) in the age groups of 25–34 and 35–45 years. According to the survey, the incidence of thyroid diseases in the studied groups is comparable. Every fifth woman indicated menstrual irregularities. Among women 25–45 years old, women with low-normal PRL values (Me = 4.49 [3.52; 5.41] ng/ml) have more unfavorable metabolic indicators. Metabolic syndrome (MS) was detected in 28%,with a predominant increase in the frequency of abdominal obesity — 55%, hypercholesterolemic LDL — 63%. Women with high PRL (Me = 41.35 [34.78; 45.88] ng / ml) also have an unfavorable metabolic profile: MS was detected in 47%, abdominal obesity — 56%, hypertension — 39%.Conclusions: In women 25–45 years old, low and high PRL values are more often associated with metabolic ill health. PRL values are from 7.8 to 28 ng / ml, i.e. conditionally defined as normal, highly normal and at the level of moderate hyperprolactinemia contribute to the maintenance of a favorable metabolic profile. When deciding on the treatment of women with non-tumor etiology hyperprolactinemia, it is important to assess the metabolic status, expanding their understanding of PRL as a hormone associated only with lactation and with the pituitary-gonad axis

Oxidative and antioxidant changes in blood of young people with premature coronary artery disease and abdominal obesity

Aim. To study the oxidative and antioxidant blood profile in young people with premature coronary artery disease (CAD) and abdominal obesity (AO).Material and methods. The study included 169 people. The main group consisted of 47 patients diagnosed with premature (<45 years of age) CAD, while 22 of them with AO and 25 without AO. The control group included 122 people without CAD, comparable with the main group in sex, age and body mass index. Among them, there are 67 people with AO and 55 people without AO. In all examined patients, the blood levels of total cholesterol, triglycerides, high-density lipoprotein cholesterol, MDA-modified oxidized high-density lipoprotein, superoxide dismutase (SOD), glutathione peroxidase 1, and whole blood total antioxidant capacity (TAC) were determined. Statistical processing was carried out in the SPSS 13.0 program.Results. The blood TAC was 2,3 times lower in persons with CAD than in those without CAD, and amounted to 1,58 mmol of trolox. The blood level of SOD in patients with CAD was 1,16 times higher than in the group of people without CAD. The relative odds of premature CAD were associated with decreased TAC (B=-2,855; Exp(B)=0,058; 95% confidence interval (CI), 0,008-0,392; p=0,004) and increased level of SOD (B=0,105; Exp(B)=1,076; 95% CI, 1,031-1,196; p=0,006), regardless of sex, age, risk factors. Conclusion. Thus, an increased SOD and reduced TAC can be potential biomarkers for premature CAD in people under 45 years of age

Calcification markers and long-term outcomes of coronary artery bypass grafting

Aim. To assess the long-term outcomes of coronary artery bypass grafting (CABG) and their association with calcification biomarkers.Material and methods. The study included 129 men (mean age, 61,5±7,5 years) with coronary atherosclerosis who were admitted for CABG surgery. Patients were divided into 2 groups: with favorable and unfavorable (death, myocardial infarction, stroke, surgery) 5-year prognosis after surgery. Before the surgery, the blood concentrations of calcification biomarkers (osteoprotegerin, osteopontin, osteonectin and osteocalcin) were determined in all patients.Results. Long-term outcomes of myocardial revascularization were studied in 92 patients (71%). An unfavorable long-term 5-year period was identified in 28 men (30,4%). In men with an unfavorable 5-year prognosis, the blood osteocalcin level before CABG was 1,2 times higher than in men with a favorable one. Multivariate linear regression showed that the risk of a 5-year unfavorable prognosis for coronary atherosclerosis after myocardial revascularization was associated with the blood osteocalcin concentration, determined before CABG (B=0,018, R2=0,285, p=0,008).Conclusion. The data obtained indicate the relevance of continuing studies on osteocalcin, including with respect to its contribution to coronary atherosclerosis and calcification

The frequency of metabolically healthy and unhealthy phenotypes in women aged 25–44 years with different levels of thyroid-stimulating hormone, prolactin, leptin

Reports that the risk of developing cardiovascular diseases in obesity is not the same, led to the allocation of metabolically healthy and unhealthy phenotypes (MHP and MUHP), this concept is based on the ability of adipose tissue to produce a number of adipokines, one of which is leptin. Hormones such as thyroid-stimulating hormone (TSH) and prolactin (PRL) are interesting from the point of view of their effect on metabolism. Aim of the study was to investigate the prevalence of MHP and MUHP in young women with different body mass index (BMI) and hormonal status (TSH, PRL, leptin) in different phenotypes. Material and methods. A group of women (n = 655) was selected from a representative sample of the Novosibirsk population aged 25–44 to study clinical and laboratory parameters. The design is a cross – sectional, observational, single – centre study. IDF, 2005 and NCEP ATP III, 2001 criteria were used to evaluate the MHP and MUHP. Results and discussion. The prevalence of MUHP in young women in Novosibirsk was 22.3 %, MHP – 77.7 % according to IDF, 2005; according to NCEP ATP III, 2001 – 13.1 and 86.9 %, respectively. The prevalence of MUHP increased with increasing BMI from 4.0 to 72.0 % according to IDF, 2005 and from 2.3 to 58.0 % according to NCEP ATP III, 2001, respectively. In obese women, MUHP was detected twice as often as MHP – 72 and 28 % according to IDF, 2005, 58 and 42 % according to NCEP ATP III, 2001. TSH and PRL do not provide information about metabolic health in young women. Leptin content is associated with BMI. The threshold value of the leptin level was 18.3 ng/ml with maximum sensitivity and specificity (Se = 53.3 %, Sp = 81.5 %), 14.5 ng/ml with equal sensitivity and specificity (Se = 65.7 %, Sp = 65.7 %). The area under ROC curve (AUC) for diagnosis of MUHP was 0.727 (SE = 0.029, p < 0.0001). Conclusion. The frequency of MHP in young women decreases with increasing BMI. MUHP is 3.5 times less common than MHP according to IDF criteria, 2005. TSH and PRL are not associated with the metabolic phenotype in young women. A leptin level more than 18.3 ng/ml has been identified as one of the markers for the recognition of MUHP in women aged 25–44 years, regardless of BMI

Changes induced in mouse lipid metabolism by simultaneous impact of antisense oligonucleotide derivatives to <i>apoB</i>, <i>PCSK9</i>, and <i>apoCIII</i> mRNAs

Development of new drugs able to decrease the level of “bad” cholesterol, in particular, based on antisense oligonucleotide derivatives (ASOs), remains relevant for the patients with familial hypercholesterolemia and/or intolerant to statins. The goal of the work was to assess the changes in the lipid metabolism caused by variants of joint impact of the ASOs targeted to the mRNAs of its key genes: apoB, PCSK9, and apoCIII. Female C57BL/6J mice; nuclease-protected 13- and 20-nucleotide ASOs, and standard protocols for quantification of lipoproteins (HDL CHL, non-HDL CHL, and total CHL) and ALT in the blood serum were used in the work. The following combinations of ASOs were four times injected to the mouse caudal vein: 1) ASO to apoB, 2) ASO to apoCIII, 3) ASO to apoB and ASO to PCSK9, 4) ASO to apoB, ASO to PCSK9, and ASO to apoCIII, 5) ASO to apoB (three doses), ASO to PCSK9, and ASO to apoCIII (two doses), 6) ASO to PCSK9 and (ASO to apoCIII – only in the fourth administration). Triple weekly administration of these ASO combinations resulted in a decrease in non-HDL CHL by 25, 16, 35, 47, 60, and 7 %, respectively, as compared with the control and 1.8-, 1.5-, 1.9-, 2.4-, 3.1, and 1.24-fold higher HDL CHL/ non-HDL CHL ratio. The subsequent ASO injection with concurrent switching to a high-fat diet after 1 week resulted in a decrease in the non-HDL CHL by 28, 2, 28, 70, 33, and 49 % for ASOs (1–6), respectively, as compared with the control; the HDL CHL/non-HDL CHL ratio was 1.5-, 1.1-, 2-, 3.7-, 1.9-, and 2-fold better. The ALT concentration for all ASO combinations remained within the norm for the control animals, demonstrating the absence of any hepatotoxic effect. The best efficiency of ASOs requires selection of concentrations for single ASOs and their combinations as well as of the order and timing of administration. Thus, a new antisense approach is proposed

NEW ASPECTS OF THE USE OF PROTEIN TISSUE-SPECIFIC MARKERS IN THE ESTIMATION OF SEVERITY OF COMMUNITY-ACQUIRED PNEUMONIA

The purpose of the study was to assess the associations of proteins of pulmonary surfactants A (SP-A) and D (SP-D) with the severity of community-acquired pneumonia. Material and methods. A survey of 188 patients diagnosed with community-acquired pneumonia (CAP). The average age of the examined patients was 54.3 ± 16.5 years (M ± SD). In 102 patients (54.3 %) severe CAP was verified. All patients underwent clinical, functional, diagnostic and laboratory studies. Results and discussion. It was determined that SP-A and SP-D content was significantly higher in patients with severe CAP compared to patients with a mild course of this disease. The direct association of severe CAP with an elevated SP-AP level, directly associated with an increase in the concentration of C-reactive protein in the blood and erythrocyte sedimentation rate, have been confirmed in early studies. Using partial correlation analysis, it was established that the severe course of CAP is directly related to SP-A (r = 0.221; p = 0.003) and SP-D content (r = 0.262; p &lt; 0.001) regardless of age, smoking, and body weight. Thus, direct associations of SP-A and SP-D with a severe course of CAP reflect the high pathogenetic significance of these protective factors in infectious lung damage