22 IN T E R N A L m E d Ic IN E ISSN 2413-6077. IJmmR 2017 Vol. 3 Issue 1 dOI 10.11603/IJMMR.2413-6077.2017.1.7867 ThE EffEcT Of L-ARGININE ON OXIdATIVE sTREss ANd mIcROALBUmINURIA IN PATIENTs WITh TYPE 2 dIABETEs mELLITUs ANd chRONIc KIdNEY dIsEAsE L. P. Martynyuk, L. Z. Vons, O. O. Ruzhytska I. HORBACHEVSKY TERNOPIL STATE MEDICAL UNIVERSITY, TERNOPIL, UKRAINE Background. Diabetic kidney disease (DKD) is one of the severest complications of diabetes. Microalbumi- nuria (MAU) is one of the first signals of DKD and an important pathogenetic mechanism of disease progression. With diabetes antioxidant properties worsen significantly. Objective. The aim was to investigate the effect of L-arginine on oxidative stress parameters and microal- buminuria in type 2 diabetes mellitus and chronic kidney disease patients. Methods. In total, 57 patients with type 2 diabetes mellitus and chronic kidney disease and 30 healthy in- dividuals (control group) took part in the study. The patients were divided into 2 congruent groups. The 1-st group of patients (n=33), in addition to standard therapy, received L-arginine 4.2 g intravenously for 5 days, after that they took it 1.0 g orally three times a day during meals for 1 month. The second group of patients (n=24) received a standard therapy. The concentration of lipid peroxidation products was measured by a spectrophotometric method. The de- termination of MAU was carried out in morning portion of urine immunological semiquantitative using test strips. Results. Significant improvement in indexes of lipid peroxidation was observed in both groups after thera- py (p˂0.01), but in patients treated with L-arginine it was more expressed (p˂0.01). The standard therapy did not significantly affect the level of MAU (p˃0.05). The patients treated with L-Arginine, showed a significant reduction in MAU (p˂0.01). Conclusions. L-arginine facilitates the correction of lipid peroxidation processes and reduces the severity of microalbuminuria in patients with diabetic kidney disease that slows down its progression. KEy wORDS: diabetes mellitus, chronic kidney disease, lipid peroxidation, microalbuminuria, L-arginine. Corresponding author: Liudmula Vons, Department of Internal Medicine №3, I. Horbachevsky Ternopil State Medical Uni- versity, 1 Clinichna Street, Ternopil, Ukraine, 46002 Phone number: +380965406768 E-mail: vonsluyda@gmail.com Introduction In recent decades a number of patients with diabetes mellitus (DM) is increased, the patients with type 2 diabetes mellitus are a significant share of them [1, 2, 3]. Diabetic kidney disease (DKD) is one of the severest complications of diabetes; it is regis- tered in 20–30% of patients with diabetes. Among the patients, who receive renal replace- ment therapy, the share of people with DKD is about 40–50% [2, 4]. It is established that 10- 20% of patients with diabetes die from chronic renal failure [4, 5]. Microalbuminuria (MaU) is one of the first signals of DKD. It is not only the symptoms but also an important pathogenetic mechanism of progression [6, 7]. According to recent studies, MAU is considered to be an independent predic- tor of progressive renal disease [6, 7]. Despite the fact that MAU is regarded as a risk factor, in the timely diagnosis and adequate treatment it can achieve the level of recourse normoalbu- minuric [6, 8]. But in 30–45% of patients over 10 years old the disease progresses to pro- teinuria. On the other hand, in 20% of patients over 20 years old a terminal kidney failure is developing either [6, 7]. With diabetes antioxidant properties wors- en significantly. the activity of antioxidant de- fence in the tissues is reduced; a lipid peroxida- tion is disordered that has repeatedly been proven in experimental diabetes [9, 10]. It is important to reveal as soon as possible the disorders of pro- and antioxidant system for diagnosis and monitoring of the health state. The timely correction of changes can prevent the progression of vascular complications of diabetes [9]. International Journal of Medicine and Medical Research 2017, Volume 3, Issue 1, p. 22–25 copyright © 2017, TSMU, All Rights Reserved L. P. Martynyuk et al. 23 IN T E R N A L m E d Ic IN E ISSN 2413-6077. IJmmR 2017 Vol. 3 Issue 1 The aim of our research was to investigate the effect of L-arginine on oxidative stress pa- rameters and microalbuminuria in type 2 dia- betes mellitus and chronic kidney disease pa- tients. Methods A total of 57 patients with type 2 diabetes mellitus and I-V stages of chronic kidney dis- ease took part in the study. There were 27 (47%) men and 30 (53%) women among them. The average age of the patients was (58.7±1.2) years old. The average duration of diabetes is (11.4±0.9) years. The average illness period of CKD is (2.5±0.3) years. The average weight of patients was (81.7±1.4) kg. 30 healthy individu- als composed the control group. The diagnosis was established by the Sec- ond National Congress which was adopted as nephrology classification of diseases of the urinary system in 2005. The stage of CKD was determined by glomerular filtration rate (gfR), defined by the formula CKD-EPi [11]. The concentration of thiobarbituric acid reactive substances (TBARS) was determined in the blood serum by a spectrophotometric me- thod [12]. We also evaluated the concentrations of glutathione (SH-group) [13], catalase [14], superoxide dismutase (SOD) [15] in the blood plasma. The determination of MAU was carried out in morning portion of urine immunological semiquantitative using test strips CLINITEK® Microalbumin 2 Strips (Germany) by means of the unit CLINITEK Status® + Analyser (Germa- ny). MAU was diagnosed at the level of urinary albumin excretion within 30–300 mg/day, nor- moalbuminuria – less than 30 mg/day. The patients with DKD were divided into 2 congruent groups: the 1-st group of patients (n=33), who, in addition to standard therapy received L-arginine 4.2 g intravenously for 5 days, after that they took it 1.0 g orally three times a day during meals for 1 month. The second group of patients (n=24) received a standard therapy. The results were compared with those of the healthy controls (n 30). The data were subjected to statistical re- search processing. For this purpose the applica- tion package Statistica (StartSoft USA, v.12) was used: the method of parametric and nonpara- metric statistics, Spearman rank correlation to determine the connection between the studied parameters. Statistically significant differences in p˂0.05 are considered. Results Analyzing the results, it was found outthat 100% of patients with DKD have lipid peroxi- dation disorders (Fig. 1). The average level of MAU in the DKD pa- tients before treatment is in 25.6 times higher than in the control group (Fig. 2). We have analysed the findings of MAU, depending on the method of treatment in DKD patients groups before and after treatment (Table 2). Discussion The average levels of TBARS and SH-groups in the patients with DKD before treatment were significantly increased compared with the cor- responding values in the control group (p˂0.01). Significantly reduced SOD and catalase activity were detected in the observed DKD patients before treatment compared with the control group (p˂0.01). no significant difference in lipid peroxidation indexes was noted between two DKD patients groups before treatment (р˃0.05). Significant improvement in lipid peroxidation indexes was observed in both DKD patients groups after therapy (p˂0.01), but in the pa- Fig. 1. The level of lipid peroxidation indexes in DKD patients before treatment and control groups. * – The difference between the control group and DKD patients group is significant (р˂0.01). Fig. 2. The level of MAU in the DKD patients before treat- ment and in the control groups. * – The difference between the control group and the DKD patients group is significant (р˂0.01) L. P. Martynyuk et al. 24 IN T E R N A L m E d Ic IN E ISSN 2413-6077. IJmmR 2017 Vol. 3 Issue 1 tients treated with L-arginine it was more ex- pressed (p˂0.01). the standard therapy did not significantly affect the level of MaU (p˃0.05). a significant reduction in MaU (p˂0.01) was proved in the patients treated with L-Arginine. The role of oxidative stress in the pathogen- esis of diabetes has been examined repeat- edly in the experimental models. Renal cortical superoxide production is increased in the early stage of experimental diabetes that leads to vasoconstriction of afferent arteriole [16]. Recent studies in experimental models of dia- betic nephropathy indicate that vascular syn- thesis of NO protects from progression of renal lesions in diabetes [17]. L-arginine is a semi- essential amino acid and also the main source for the generation of NO via nitric oxide syn- thase [18]. Recent study demonstrated that L-arginine supplementation in type II diabetic rats was beneficial by preserving glomerular filtration rates, presumably via increased renal endothe- lial nitric oxide synthase levels, that leads to renal vasodilation [19]. In our present research L-arginine supple- mentation, which stimulates nitric oxide syn- thesis, causes a pronounced improvement in lipid peroxidation indexes and a significant re duction in MAU in the patients with DKD. Conclusions In patients with DKD, the decrease in activity of SOD and catalase and the increase of TBARS The effects of treatment on lipid peroxidation indexes are presented in Table 1. Table 1. The dynamics of lipid peroxidation indexes in DKD patients before and after treatment depending on different treatment regimens, M±m Indexes Groups p group І (n=33) group ІІ (n=24) p1 p2 p3 TBARS, mkmol/l Before treatment 6.26±0.20 6.20±0.13 р˃0.05 After treatment 3.81±0.27 5.27±0.46 р˂0.01 р˂0.01 р˂0.05 SOD, % Before treatment 41.76±0.68 41.45±0.66 р˃0.05 After treatment 55.61±1.42 43.21±1.18 р˂0.01 р˂0.01 р˃0.05 SH-groups, mlmol/l Before treatment 42.59±1.23 43.71±2.08 р˃0.05 After treatment 57.05±1.98 51.14±1.65 р˂0.05 р˂0.01 р˂0.05 Catalase, % Before treatment 53.74±2.16 51.46±1.18 р˃0.05 After treatment 31.39±2.80 47.25±2.88 р˂0.01 р˂0.01 р˃0.05 notes: 1. p1 – significant difference in performance between groups before and after treatment. 2. p2 – significant difference in performance in the first group before and after treatment. 3. p3 – significant difference in performance in the second group before and after treatment. Table 2. The dynamics of MAU levels, depending on different treatment regimens, M±m МАU mg/day Groups p group І (n=23) group ІІ (n=24) p1 p2 p3 Before treatment 180.7±19.9 188.6±15.6 р˃0.05 After treatment 51.4±15.9 149.2±15.5 р˂0.01 р˂0.01 р˃0.05 notes: 1. p1 – significant difference in performance between groups before and after treatment. 2. p2 – significant difference in performance in the first group before and after treatment. 3. p3 – significant difference in performance in the second group before and after treatment. L. P. Martynyuk et al. 25 IN T E R N A L m E d Ic IN E ISSN 2413-6077. IJmmR 2017 Vol. 3 Issue 1 and SH-group parameters is observed that proved a lipid peroxidation disorder. L-arginine in the standard therapy facilitates the correction of lipid peroxidation processes and reduces the severity of microalbuminuria in patients with diabetic kidney disease that slows down its progression. References 1. Schieppati A. 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