The study aimed to assess serum Klotho protein level in type 2 diabetic patients depending on kidney function. Methods. This observational study included 72 patients with diabetes mellitus (DM) and 26 patients with acute coronary syndrome. The control group consisted of 20 healthy subjects. Depending on the presence of albuminuria and glomerular filtration rate (GFR), the diabetics were divided into the following groups: group I included the patients with normal GFR and without albuminuria (n = 25); group ІІ consisted the patients with normal GFR and albuminuria (n = 23); group III – the patients with reduced GFR and albuminuria (n = 24) and group ІV included the patients with acute coronary syndrome (n = 26). The GFR was calculated using the CKD EPI formula (KDIGO 2012). The concentration of Klotho protein was determined by enzyme-linked immunosorbent assay. Results. The development of diabetic nephropathy in type 2 diabetic patients accompanied by a significant decrease of soluble Klotho compared with the controls and the patients of the1-st group. The level of Klotho protein in the group of patients with albuminuria decreased to (490.66 ± 58.76) pg/ml (p <0.05). The lowest concentration of Klotho (443.58 ± 46.92) pg/ml was found in the advanced stages of diabetic nephropathy, namely in patients with albuminuria and impaired renal function. Moreover, a significantly decreased serum Klotho was observed in acute coronary syndrome group in comparison with the control group (p <0.05). There were inverse correlations of Klotho concentration with urinary albumin and blood creatinine levels and a direct correlation between Klotho and GFR. Conclusions. The obtained data indicated the key role of Klotho protein in the formation of renal pathology in type 2 diabetes and the feasibility of practical use of Klotho determination as an early diagnostic marker of renal disorders and cardiovascular risk assessment. The strategies improving Klotho production may be useful in the reduction of both renal and vascular lesions progression in type 2 diabetic patients.

Abstract. The present study aimed to investigate the dynamics of clinical and laboratory parameters and serum Klotho protein level in patients with diabetic kidney disease using nephroprotective therapy combined with an inhibitor of the sodium-glucose cotransporter 2 (SGLT2) dapagliflozin. Methods. A total of 76 type 2 diabetic patients with diabetic nephropathy (DN) were examined in this prospective study. Control group - 20 healthy subjects. 53 patients received a standard course of treatment, which included metformin, renin-angiotensin-aldosterone system blockers and statins. In addition to standard therapy, 23 patients have been prescribed the SGLT2 inhibitor dapagliflozin 10 mg per day. The treatment follow-up period was six months. Klotho concentration was determined by an enzyme-linked immunosorbent assay. Results. The development of DN in type 2 diabetic patients was accompanied by a significant decrease in soluble Klotho protein in comparison with controls and patients without nephropathy. During follow-up, Klotho protein level was changed significantly in the group of DN patients with albuminuria. Standard therapy resulted in Klotho concentration increase by 14% compared to pre-treatment values; a more demonstrative increase in the Klotho level was found in the dapagliflozin group (almost 23%). Conclusions. SGLT2 inhibitor treatment resulted in a significant increase of pleiotropic serum protein Klotho in patients with type 2 diabetes and diabetic kidney disease.

Objective — to determine the prevalence of the Alu Ins/Del polymorphism of the angiotensin‑converting enzyme (ACE) gene in patients with type 2 diabetes mellitus (DM 2) with nephropathy and to identify possible associative relationship between the course of the disease and the genetic profile of the examined subjects. Materials and methods. Examinations involved 73 patients with diabetic nephropathy (DN), treated in the hospital of L. T. Mala Therapy National Institute. The control group consisted of 19 healthy individuals. After the initial examination and depending on the polymorphic variant of the ACE gene, patients were divided into three groups: group I — carriers of D/D polymorphism of the ACE gene (n = 23); group II — patients with I/D polymorphism of the ACE gene (n = 32); Group III — patients with I/I polymorphism of the ACE gene (n = 18). DNA amplification and genotyping were carried out by real‑time polymerase chain reaction (PCR) using the SNP‑Express‑Shot reagent kit (Litech, RF) and the CFX96 touch real‑time PCR product detection system (BioRad). Results. The distribution of the genotypes of the Alu I/D polymorphism of the ACE gene corresponded to the Hardy‑Weinberg equilibrium in all studied groups and did not differ significantly from European populations. Comparison of patients with DN, carriers of different Ins/Del polymorphic variants of the ACE gene, did not show significant differences in the stages of chronic kidney disease, creatinine level, and glomerular filtration rate (GFR). Patients with type 2 DM, homozygous for the I/I allele of the ACE gene, had significantly lower levels of urinary albumin (16.80 [12.61 — 34.20] mk/ mL), compared with I/D heterozygotes (26.07 [20.91 — 44.27] mkg/mL), p < 0.05, which demonstrates the negative effects of D allele on the kidney damage progression in this category of patients, even under the same GFR. Conclusions. Patients with DN who are homozygous for the ACE gene allele I, have significantly lower albumin levels than I/D heterozygotes, demonstrating an association between the D allele and the progression of renal disease in type 2 DM. Determining the ACE gene polymorphism in patients with type 2 DM allows to identify risk groups for the DN development and will help to make individualized adjustments to the scheme of pharmacological therapy.

Diabetic kidney disease develops in approximately 40% of patients who are diabetic and is the leading cause of chronic kidney disease worldwide. The majority of patients actually die from cardiovascular diseases and infections before needing kidney replacement therapy. The progression of fibrotic processes in the diabetic kidneys occurs with the participation of universal mediators of inflammation. Given the clinical and social significance of diabetic kidney damage, the determination of interleukin-1β and interleukin-6 in blood serum may expand the understanding of this problem, as well as help in the development of individualized therapeutic approaches. The purpose of the study is to investigate interleukin-1β and interleukin-6 levels in the serum of patients with type 2 diabetes mellitus, depending on kidney functional state in the dynamics of therapy and with additional prescription of dapagliflozin. Materials and methods. 72 patients with type 2 diabetes were examined. The control group consisted of 20 healthy individuals. The patients were divided into the following groups: group I – type 2 diabetic patients with normal glomerular filtration rate and without albuminuria (n = 25); group II – patients with type 2 diabetes with normal glomerular filtration rate and albuminuria (n = 23); group III – patients with type 2 diabetes with decreased glomerular filtration rate and albuminuria (n = 24). The complex pathogenetic therapy of patients of group 1 included renin-angiotensin-aldosterone system blockers, metformin and hypolipidemic agents. Patients of group 2 were additionally prescribed dapagliflozin. The control was carried out after 6 months. Results and discussion. According to the results of the study, the development of diabetic nephropathy in patients with type 2 diabetes is accompanied by a significant increase in the level of interleukin-1β and interleukin-6 in the blood plasma compared with the control and in patients with diabetes without signs of nephropathy. Nephroprotective therapy contributed to a significant decrease of interleukin-1β and interleukin-6 levels in the blood serum in all groups of patients. The highest response to treatment was observed in patients with diabetic nephropathy and albuminuria. Most significant decrease of albuminuria, normalization of blood pressure and lipid spectrum improvement were observed in patients with the initial stages of the disease. Additional using of dapagliflozin leads to a more significant decrease in the content of proinflammatory cytokines in the blood serum of patients with type 2 diabetes mellitus with diabetic nephropathy. Conclusion. The results of the study indicate the advisability of using interleukin-1β and interleukin-6 as a diagnostic marker of cardiac disorders, assessment of prognosis, and improvement of the cardionephroprotective strategy in diabetic patients. The therapeutic potential of dapagliflozin in the treatment of diabetic nephropathy and the prevention of cardiovascular events in patients with diabetes is of great scientific interest and requires further research

Objective — to reveal an association of I/D polymorphism of ACE gene with metabolic factors of cardiovascular risk (CVR) such as visceral obesity (VO), insulin resistance (IR), dyslipidemia (DLP) as well as with indices characterized structural and functional state of left cardiac chambers in patients (pts) with diabetic nephropathy (DN) and essential hypertension (EH). Materials and methods. The investigation was conducted at the Clinical Department of Arterial Hypertensions and Kidney Diseases in L. T. Mala National Therapy Institute of NAMS of Ukraine (Kharkiv). Clinical examinations involved 82 pts, 42 (51.2 %) females and 40 (48.8 %) males with DN of I — IV stage and EH of II — III stage, aged 31 to 82 years old (an average age is (61.65 ± 1.28) years old). Three genotypes of I/D polymorphism of ACE gene included II — 18 (24.7 %), ID — 32 (43.8 %) and DD — 23 (31.5 %) of cases correspondingly were investigated in 73 (89.0 %) pts. Anthropometric measurements with calculations of body mass index (BMI), body fat percentage (BFP), body fat mass (BFM), fat mass index (FMI) were conducted by known formulas in all pts. Blood lipids were detected by enzyme method with measuring of total cholesterol, high‑density lipoprotein cholesterol (HDL‑C) and triglycerides (TG). Levels of cholesterol of very low density and low‑density lipoprotein (VLDL‑C, LDL‑C) were calculated by standard formulas. Blood glucose level was measured by glucose oxidase method and serum insulin was detected by immunoassay method. IR indices such as HOMA‑IR, triglyceride glucose index (TGGI) and METS‑IR (metabolic score for insulin resistance) were obtained by known formulas. I/D polymorphism of ACE gene (rs 4646994) was investigated with a usage of polymerase chain reaction. Structural and functional state of left cardiac chambers was estimated using transthoracic echocardiography. Systolic and diastolic blood pressure (SBP and DBP) were measured by Korotkov’s method. The results were statistically obtained with computer programs Microsoft Office Excel 2003 and Statistica 23.0. Results. In pts with II genotype, LDL‑C levels positively correlated with BFP (r = 0.490; p = 0.046) and BFM (r = 0.484; p = 0.049) in conditions with hypertriglyceridemia due to activation of visceral fat lipolysis in which a reverse correlation between relative thickness of left ventricle wall (RTLVW) and serum TG concentration (r = –0.540; p = 0.02) could be explained. In pts with ID genotype, presence of D allele in genotype was associated with an increase in RTLVW due to SBP influence (r = 0.358; p = 0.045). In genotype ID there was a relationship between IR index HOMA‑IR and BMI (r = 0.399; p = 0.029) and HOMA‑IR and FMI (r = 0.402; p = 0.025) that demonstrated VO participation of in the development of IR in pts with DN and EH and genotype ID. In pts with DN and EH, who had DD genotype, SBP level reversely correlated with HDL‑C (r = –0.498; p = 0.018) that could be explained by influence of IR on elevation of SBP and reduction of HDL‑C. In pts with DD genotype both SBP and DBP almost equally influenced on the left atrial diameter (r = 0.460; p = 0.036 and r = 0.453; p = 0.034 correspondingly). In pts with DN and EH whose had DD genotype IR index METS‑IR positively correlated with RTLVW (r = 0.419; p = 0.047) and left ventricle myocardial mass index (r = 0.518; p = 0.011) that confirmed an association of D allele of ACE gene with left ventricle hypertrophy caused both by EH and IR. Conclusions. It has been established that in patients with diabetic nephropathy and essential hypertension, the I/D polymorphism of ACE gene was associated with such metabolic factors of cardiovascular risk as visceral obesity, insulin resistance, dyslipidemia, and pathological left ventricle remodeling.