Oct 6, 2014 - Bulletin of Experimental Biology and Medicine, Vol. 157, No. ... duction, Siberian Division of the Russian Academy of Medical Sci- ences, Irkutsk ...
DOI 10.1007/s10517-014-2649-z Bulletin of Experimental Biology and Medicine, Vol. 157, No. 6, October, 2014
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GENERAL PATHOLOGY AND PATHOPHYSIOLOGY Integral Indicator of Oxidative Stress in Human Blood L. I. Kolesnikova, N. V. Semyonova, L. A. Grebenkina, M. A. Darenskaya, L. V. Suturina, and S. V. Gnusina
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 157, No. 6, pp. 680-683, June, 2014 Original article submitted February 14, 2014 Here we performed individual evaluation of the oxidative stress index that serves as an integral criterion for the balance of LPO–antioxidant defense system in women with endocrine pathology (type 1 diabetes mellitus and infertility with hyperprolactinemia). The state of the LPO–antioxidant defense system was estimated from blood levels of LPO substrates with conjugated double bonds, conjugated dienes, ketodienes, conjugated trienes, thiobarbituric acid-reactive substances, retinol, α-tocopherol, reduced and oxidized glutathione, and SOD activity. The use of this oxidative stress index allowed us to diagnose oxidative stress in female patients with endocrine pathology. Key Words: oxidative stress index; diabetes mellitus; infertility Pathogenesis of neuroendocrine diseases that are highly prevalent in humans is closely related to nonspecific biochemical processes, including LPO in cells and in the body, determining reactivity and adaptive capacities of the body [9]. Activation of free radical processes and decrease in the buffer capacity of antioxidant defense (AOD) lead to oxidative stress, one of the pathogenetic stages in various endocrine diseases (e.g., type 1 and 2 diabetes mellitus (DM) [3] and endocrine infertility [4,6]) accompanied by changes in the protein, lipid, carbohydrate, and nucleic acid metabolism and salt-water balance [8,10]. Integral criteria are extensively used in clinical practice for detection of pathological states and diagnosis of a variety of disturbances caused by environmental factors. These criteria are more sensitive to imbalance in the LPO–AOD system than individual parameters. The complexity and multicomponent naResearch Center for Problems of Family Health and Human Reproduction, Siberian Division of the Russian Academy of Medical Sciences, Irkutsk, Russia. Address for correspondence: natkor_84@ mail.ru. N. V. Semyonova
ture of the LPO–AOD system hamper quantitative analysis of oxidative stress and prediction of the biological response and therapeutic effect, which complicates interpretation of the results and antioxidant treatment. Oxidative stress index (OSI) was proposed to be evaluated from the prooxidant/antioxidant ratio in patients with different pathologies [1,5,7]. Here we performed individual evaluation of OSI in female patients with endocrine pathology (DM1 and infertility with hyperprolactinemia). OSI was used as an integral systemic criterion for abnormalities of the LPO–AOD system [2].
MATERIALS AND METHODS Two endocrine disorders (DM1 in reproductive-age women and female infertility with hyperprolactinemia) were used as the models to study oxidative stress and reliability of OSI. The inclusion criteria for women with DM1 (main group, N=15; mean age 35.0±3.4 years, DM1 history 12.8±2.0 years) were verified diagnosis of DM1 and insulin therapy. The exclusion criteria for the main
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Bulletin of Experimental Biology and Medicine, Vol. 157, No. 6, October, 2014 GENERAL PATHOLOGY AND PATHOPHYSIOLOGY
group were severe associated somatic diseases and severe manifestations of diabetic complications (chronic renal failure and microangiopathy). We used WHO classification and diagnostic criteria of DM (1999) and National Standards for the Care of DM Patients (2002). The control group consisted of 20 women (mean age 28.2±1.5 years) without DM1, acute disorders, or exacerbations of chronic diseases. The inclusion criteria for women with infertility and hyperprolactinemia (main group, N=60; mean age 26.0±5.6 years) were an increase in serum prolactin concentration >600 U/ml during the follicular phase of the menstrual cycle (exceeding the reference level in at least 2 cycles); absence of pregnancy in regular sex life without contraception for at least 1 year; disturbances in the menstrual cycle (oligomenorrhea in anovulation or luteal phase defect; or luteal phase defect with regular menstruation); and galactorrhea. Magnetic resonance tomography of the pituitary gland was performed to exclude/confirm organic diseases of the pituitary gland or boundary regions (pituitary micro- and macroadenoma, empty sella syndrome, and CNS tumors). The exclusion criterion for the main group was severe concomitant somatic pathology. The control group consisted of 61 fertile women (mean age 22.0±2.3 years) without neuroendocrine disorders. Their case history included pregnancy with delivery over the last 2 years. Control group women were characterized by regular menstrual cycle and had no severe somatic diseases. All subjects signed informed consent form. The study was performed according to the ethical principles of the World Medical Association Declaration of Helsinki (2000). The intensity of LPO and state of AOD were evaluated from the content of their individual components in blood serum and hemolysate. The amount of LPO
substrates and products (compounds with conjugated double bonds, conjugated dienes, ketodienes, and conjugated trienes) was measured spectrophotometrically. The concentration of thiobarbituric acid-reactive LPO products was estimated fluorometrically in the reaction with thiobarbituric acid (TBA). Activity of AOD system was evaluated by the contents of α-tocopherol, retinol, and reduced and oxidized glutathione. SOD activity was determined by the dynamics of epinephrine autooxidation. The measurements were performed on a Shimadzu RF-1501 spectrophotometer. The results were analyzed with Statistica 6.1 software (StatSoft). Between-group differences for independent samples were evaluated by parametric Student’s test and nonparametric Mann–Whitney test at critical significance level of 5%.
RESULTS The content of LPO substrates in women with both pathological states was elevated by 51% (DM1, p
