Yerbolat Iztleuov¹, Talgar Abilov², Ganiya Zhanabayeva³, Irina Ismailova⁴ and Marat Iztleuov²

¹Department of Radiation diagnostics, Faculty of Medicine, West–Kazakhstan Marat Ospanov State Medical University, Aktobe, Kazakhstan.

²Department of Natural sciences, Faculty of Medicine, West–Kazakhstan Marat Ospanov State Medical University, Aktobe, Kazakhstan.

³Department of General dentist, Faculty of Stomatology, West–Kazakhstan Marat Ospanov State Medical University, Aktobe, Kazakhstan.

⁴Department of General medical practice, Faculty of Medicine, West–Kazakhstan Marat Ospanov State Medical University, Aktobe, Kazakhstan.

Corresponding Author E-mail: ermar80@mail.ru

DOI : https://dx.doi.org/10.13005/bpj/1367

Abstract

Hexavalent chromium presents a particular threat due to its high toxicity. In this study showed the neuroprotective effect of sodium tetraborate in chromic intoxication. The experiment was performed on Wistar rats divided into 4 groups: 1 group - control; 2 group - single intraperitoneal injection of K₂Сr₂O₇ in a dose of 0,5LD₅₀, in the 3 and 4 groups with drinking water received Na₂B₄O₇. Animals of the 3 group - 4 mg/kg of body weight, in 4 group - 72 mg/kg of body weight daily for 10 days, the last administration of Na₂B₄O₇ was combined with a single intraperitoneal injection of K₂Сr₂O₇ at a dose of 14 mg/kg (0,5LD₅₀). Na₂B₄O₇ in a dose of 4 mg/kg leads to a decrease in the level of MDA by 33%, an increase in the activity of catalase by 69%, superoxide dismutase by 21%, GR by 49%. Na₂B₄O₇ at a dose of 72 mg/kg increases the MDA content by 31% in comparison with the data of rats of chromic intoxication. K₂Сr₂O₇ reduces the GSH level by 42%, the non-protein thiol by 36%. Na₂B₄O₇ at a dose of 4 mg/kg increased the GSH level by 53%, the non-protein thiol by 35%, and at a dose of 72 mg/kg reduced the GSH content by 23% (ρ<0.05), the non-protein thiol by 20% (ρ<0.05) in comparison with the data of rats exposed to K₂Сr₂O₇.

Keywords

Antioxidant System Bichromate Potassium; Brain Damage; Lipid Peroxidation; Sodium Tetraborate;

Download this article as: 

Copy the following to cite this article: Iztleuov Y, Abilov T, Zhanabayeva G, Ismailova I, Iztleuov M. Protective Effect of Sodium Tetraborate on Chromium-induced Brain Damage in Rats. Biomed Pharmacol J 2018;11(1).

Copy the following to cite this URL: Iztleuov Y, Abilov T, Zhanabayeva G, Ismailova I, Iztleuov M. Protective Effect of Sodium Tetraborate on Chromium-induced Brain Damage in Rats. Biomed Pharmacol J 2018;11(1). Available from: http://biomedpharmajournal.org/?p=18844

Introduction

Pollution of the environment by heavy metals, as a result of high industrial activity in the late nineteenth and early twentieth centuries, has increased significantly throughout the world. Among heavy metal ions, chromium (VI) is a particular threat due to its high toxicity.¹ Ecopathogenic risk in the Western region of the Republic of Kazakhstan is associated with geochemical features – chromic biogeochemical region, oil and gas processing complex. The first in the countries of independent states and the world’s second-richest chromites ore deposit, South-Kimpersai, is located in the Aktobe region, where a large enrichment plant, chrome compounds and chromium ferroalloys are concentrated. The expansion of these industries leads to an increase in the contingent of persons having professional contact with these compounds, as well as to increase emissions to the environment, which are dangerous for the population not only of the Aktobe region but also of the border regions of Western Kazakhstan in the South Urals.²

Chromium exists in a medium in three stable oxidation states, Cr, Cr⁺³, Cr⁺⁶, which have different toxicity and transport characteristics.³ Cr⁺⁶ is considered the most toxic form, because it has a high oxidation potential, high solubility and mobility across membranes in living (systems) organisms and in the environment.⁴ The toxic effects of chromium are widely believed to be associated with the stimulation of free-radical processes, as well as the formation of intermediates during the reduction of Cr⁺⁶, which have a high reactivity.⁵ Once inside the Cr⁺⁶ cell, Cr⁺³ is reduced, generating active forms of oxygen (AFO), which cause oxidation of macromolecules such as DNA and lipids^{6,7,8,9,10,11} and induce oxidative damage to tissues such as the liver, pancreas, kidney, brain,^12,13,14,15 which has a number of negative consequences for human health, including neurotoxicity, hepatotoxicity, nephrotoxicity, genotoxicity, carcinogenicity and immunotoxicity.^16,17,18,19 And the main role in the implementation of the damaging effect of oxidative stress is played by the hydroxyl radical. The damaging role of AFO is often associated not only with their direct action on cellular structures, but with the initiation of a cascade of processes leading to cell damage.^20,21 It is well known that the brain of mammals is rich in unsaturated fat cells, has a weak antioxidant defense system and is sensitive to damage to induced free radical oxidation (FRO), mediated lipid peroxidation (LPO). Generation of FRO and enhancement of LPO of neutron membranes can lead to oxidative damage to the brain.^22,23,24,15 Reducing antioxidant activity of the brain and increasing the AFO often causes neurotoxicity.^25,26

Bohr is widely recognized as an important dietary component with numerous beneficial effects on health. Quickly absorbed from the gastrointestinal tract into the bloodstream and in physiological quantities affects a wide range of metabolic processes,^27,28,29 which is associated with antioxidant effects of boron.³⁰ Boron compounds have anti-inflammatory, hypolipidemic and antitumor actions,^31,32 are non-genotoxic³³; have a beneficial effect on the central nervous system.³⁴ Boric acid (BA) and borax are the most common form of boron in humans, which are water soluble and bioavailable.³⁵ According to a number of scientists^36,37,38,39 boric acid, borax have protective effects by modulating the indices of oxidative stress in aluminum-induced hepatotoxicity, titanium, aluminum caused by genotoxicity, thioacetamide induced hepatic insufficiency, and chloride-aluminum-induced neurotoxicity.

The results suggest that boron food supplies have beneficial effects on the central nervous system are among the most favorable assumptions that boron is a useful microelement for humans and give grounds for recommending the use of boron compounds in the violation of the CNS function.^34,40 In vivo and in vitro studies have shown that BA and boron compounds have a protective role against many cancers.^35,41 Boron carriers bind the catalytic site of the 26 S-proteasome and block its activity.^42,35 The proteasome works with a labeled protein called ubiquitin and forms the ubiquitin-proteasome pathway. This pathway plays an important role in the degradation of oxidized, mis-synthesized, damaged and unnecessary proteins in the cell-an important role in degradation of oxidized, incorrectly synthesized, damaged and unnecessary protein in the cell, and plays a key regulatory role in protein groups, involved in intracellular signaling cascades^43,44 and affect the transcription of intracellular signals of processes associated with the permeability and protection of cells and systems.^45,46,47 Consequently, the boron compounds influencing these processes can play the role of a neuroprotective agent. However, according to our information, there is no report on the effects of boron compounds on the brain under conditions of chromic intoxication, i.e. on chromium-induced neurotoxicity.

Based on the foregoing, the purpose of this study is to study the effect of sodium tetraborate on chromium-induced neurotoxicity and oxidative stress.

Materials and Methods

The work was performed on 24 male Wistar rats weighing 170-190 g. The animals were in standard conditions in the vivarium of the Central Research Laboratory of the West Kazakhstan State Medical University named after Marat Ospanov (Aktobe, Republic of Kazakhstan) on a standard diet with free access to food and water. The experiments were carried out in accordance with the European Convention for the Protection of Vertebrates used for experimental and other purposes (Strasbourg, 1986). The program of the experiment was discussed and approved by the local ethics commission of the university.

Ten days after acclimatization, animals were randomly divided into 4 groups (6 rats each): Control group: untreated animals. Experimental group 1: Animals were given a single intraperitoneal injection of potassium dichromate (K₂Сr₂O₇ “Chemistry and Technology” Ltd., Kazakhstan) at a rate of 0,5LD.^₅₀ Experimental group 2: Animals received sodium tetraborate (Na₂B₄O₇ “Farmak” Ltd., Ukraine) in drinking water at a rate of 4.0 mg/kg body weight. Experimental group 3: Animals received sodium tetraborate at a rate of 72 mg/kg of body weight for 10 days and were injected a single intraperitoneal injection of potassium dichromate at a rate of 14 mg/kg (0,5LD₅₀) at the last day.

The choice of the type of boron and chromium compounds, doses, methods of administration, and the duration of the experiment are justified by previous studies⁴⁸ and literature data.^36,37,49 Euthanization of animals in all groups was performed simultaneously 24 hours after the administration of potassium dichromate by the method of cervical instantaneous decapitation under light ether anesthesia in order to avoid stress.

The brain was rinsed in a cooled physiological solution, dewatered with filter paper, the soft marrow was removed from the blood vessels (cleaned), weighed, homogenized (10% weight/volume) in the appropriate buffer (pH-7,4) and centrifuged at 7000g for 20 minutes.

Peroxide oxidation and antioxidant status. The content of malonic dialdehyde (MDA) in the brain tissues was determined spectrophotometric by the method of Draper, Hedley.⁵⁰ The essence of the method: at a high temperature in an acidic medium, MDA reacts with 2-thiobarbituric acid, forming a colored complex with an absorption maximum at 532 nm. The molar extinction coefficient is 1,56*10^-5cm*М^-1. The MDA level was expressed in nmol/g tissue.

Catalase activity (CAT) was measured according to Koroluk et al.⁵¹ The reaction is started by adding 2 ml of hydrogen peroxide to 10 μl of the supernatant and after 10 min it is stopped by adding 1 ml of 4% ammonium molybdate. The sample absorption is measured at 410 nm. The activity of CAT is expressed in moles of H₂O₂ min/g of tissue.

The levels of non-protein thiol (NPSH) were determined by the Ellman method.⁵² An aliquot of 500μl supernatant was mixed with 10% trichloroacetic acid. After centrifugation, the protein precipitate was discarded and the free SH-groups in the clear supernatant were determined. An aliquot of 100 μl of supernatant was added to 850 μl of potassium phosphate buffer 1 M (pH=7.4) and 50 μl of 5,5-dithio-bis-2-nitrobenzoic acid – DTNB (10 mM). The colorimetric reaction was measured at 412 nm. The results were expressed as micromoles per gram of tissue (μmol/g tissue).

The level of glutathione (GSH) was determined by Ellman’s method⁵² in the modification of Jollow et al.⁵³ based on the formation of yellow staining, when DTNB (5,5-dithiobis 2-nitrobenzoic acid) is added to the sample containing SH groups. To 0.5 ml of homogenate is added to 3.0 ml of 4% sulfosalicylic acid. The mixture is centrifuged at 1600g for 15 minutes. To 0.05 ml (50 μl) of the supernatant is added the Ellmana reagent. After 10 minutes the extinction was measured at 412 nm. The amount is expressed in μg/g tissue.

The activity of glutathione reductase in the brain was determined by the method of Yawata Y., Tanaka R.⁵⁴ Principle of the method: glutathione reductase (GR) catalyzes the reduction reaction of oxidized glutathione using as the restored equivalent of NADPH₂. Reduction of the level of the latter in the test sample is determined on a spectrophotometer at a wavelength of 340 nm. The molar extinction coefficient of NADPH at E₃₄₀= 6,22мМ*см^-1. GR activity is expressed in nmol of oxidized NADPH/min/mg of protein – (nmol/min/mgPt).

The activity of superoxide dismutase (SOD) was determined by Chevari et al,⁵⁵ based on the reduction of nitrotetrazolium by superoxide radicals, which are formed by reaction with phenazinmetasulfate and the reduced form of nicotinamide adenine dinucleotide (NADH). The formation of nitroformazane, the nitrotetrazolium reduction product, is blocked in the SOD environment. The amount of nitroforman is directly proportional to the activity of SOD. The activity of SOD is expressed as a percentage.

The protein content was determined by the Lowry et al method.⁵⁶

Statistical Analysis

Statistical processing of data was carried out using the “Statistica 10” software package of StatSoft, Inc USA. Verification of the null hypothesis that there was no difference between the observed distribution was performed using the criterion of Shapiro-Wilk’s W-Test. The differences between the samples were estimated: with a normal distribution of paired variables using Student’s t-test and ANOVA in the case of multiple independent variables. The arithmetic mean values M±m of the quantitative indices, represented in the text as M±m, were calculated, where M is the average arithmetic mean, m is the error of the mean. In all statistical analysis procedures, significance level was assumed to be p<0.05.

Results

Assessment of lipid peroxidation (LPO) and non-enzymatic link of antioxidant status in the brain. Analysis of the data showed that under the influence of K₂Cr₂O₇, there is a significant increase in the LPO in t he brain, as evidenced by MDA levels (Table 1). The content of the latter is increased by 67% in comparison with the data of the control group. Preventive administration of sodium tetraborate in low dose (4 mg/kg b.w.) leads to a significant decrease in MDA level in the brain by 33%, whereas the use of Na₂B₄O₇ in a high dose (72 mg/kg b.w.) increases the MDA content by 31% with the data of rats of chromic intoxication.

Table 1: The effect of sodium tetraborate on the content of malonic dialdehyde, glutathione and non-protein thiol in the brain with chromium-induced neurotoxicity

Indicators	Groups of animals
	Control	K2Cr2O7	K2Cr2O7+ Na2B4O7 (4 mg/kg)	K2Cr2O7+ Na2B4O7 (72 mg/kg)
MDA, nmol/g	151±7.0	252±16.0x	170±140	330±19x0
GSH, μg/g	161±8.0	93±4.3x	142±7.0x0	74±3.6х0
NPSH, μmol/g	1.87±0.07	1.20±0.09x	1.62±0.11x0	0.96±0.008х0

Units:  x – p<0.05 in comparison with the control data; o – p<0.05 in comparison with the data of K₂Cr₂O₇

In addition, K₂Cr₂O₇ – intoxication significantly reduced the level in the brain GSH by 42%, non-protein thiol by 36%. Preventive use of Na₂B₄O₇ at a dose of 4 mg/kg of b.w. significantly increased the GSH level by 53% in the brain, a non-protein thiol by 35%, and the administration of sodium tetraborate at a high dose (72 mg/kg MT) reduced the GSH content by 23% (ρ<0.05), non-protein thiol by 20% (ρ<0.05) as compared to the data of rats exposed to K₂Cr₂O₇.

Enzymatic Antioxidant Status in the Cerebrum

The parameters of the enzyme link of the antioxidant system of the control and experimental groups obtained during the experiments are presented in table 2. Chromium-induced brain damage was accompanied by a significant decrease in CAT activity by 33%, GR by 25%. The activity of SOD remained at the level of control group data. Preventive use of Na₂B₄O₇ in a low dose led to a significant increase in CAT activity by 69%, SOD by 21%, GR by 49%, whereas its use in a high dose before chromic poisoning will not cause significant changes in comparison with data of animals, chromium-induced damage to the brain.

Table 2: Effect of Na₂B₄O₇ on the enzyme link of the antioxidant system of the brain with chromium-induced neurotoxicity

Indicators	Groups of animals
	Control	K2Cr2O7	K2Cr2O7+Na2B4O7 (4 mg/kg)	K2Cr2O7+Na2B4O7 (72 mg/kg)
CAT, μmol/min/mg	106±8.0	71±6.0x	120±7.00	81±6.2x
SOD, %	36±1.4	33±1.2	40±2.10	30±2.0
GR, μmol/min/mg	9.3±0.6	7.0±0.42x	10.4±0.520	7.2±0.651x

Units: x – p<0.05 in comparison with the control data; o – p<0.05 in comparison with the data of K₂Cr₂O₇

Discussion

Chromium causes a wide range of toxicological effects and physiological and biochemical dysfunctions, which involve serious health risks, including neurotoxicity.^57,18,11,15 The present study evaluated the possible consequences of different doses of sodium tetraborate (4 mg/kg and 72 mg/kg b.w.) on the central nervous system in rats exposed to potassium dichromate intraperitoneally (0,5LD₅₀ – 14 mg/kg b.w.). The brain has a relatively weak system of antioxidant protection, contains a high content of phospholipids, polyunsaturated fatty acids in comparison with other organs, consumes 20% of oxygen in the body, especially susceptible to oxidative stress and, as shown, in previous studies^58,59,13,15 Cr⁺⁶ can lead to increased LPO in the brain. As established in this study, in fact, the MDA level increased significantly under the conditions of chromic intoxication amid a significant decrease in the level of GSH and NPSH in the brain, reflecting consumption through oxidative stress. There are several ways of depletion of the level of the latter in the conditions of chromic intoxication. First, as shown by Hojo, Satomi,⁶⁰ GSH can be the source of an electron donor for the conversion of Cr⁺⁶ to Cr⁺³. Secondly, the sulfhydryl group of the cysteine ​​of the glutathione fragment has a high potency to metals, forming thermodynamic stable mercaptoid complexes with several metals. Thirdly, GSH and NPSH can be oxidized by interaction with free radicals induced by K₂Cr₂O₇.

Preventive use of tetraborate in a low dose led to a significant decrease in MDA and an increase in the content of GSH and NPSH, whereas tetraborate in a high dose, on the contrary, increased the level of MDA. An increase in the level of GSH may be due to the effect of low doses of Na₂B₄O₇ on the synthesis of de novo GSH and its regulation, or both. Therefore, in order to counteract the progressive formation of free radicals arising and leading to damage and death of brain neurons when exposed to K₂Cr₂O₇, i.e. to prevent its neurotoxicity, it is necessary to maintain the levels of GSH and NPSH.

The damaging role of FRO under the influence of Cr⁺⁶ is associated not only with their effect on cellular structures, but with the initiation of a cascade of processes leading to damage to cells and cell membranes,^20,21 including, can change the permeability of the hemoencephalic barrier (HEB) due to degeneration of endothelial cells and pericytes. On the other hand, the results obtained in this study showed that chromatin intoxication leads to a significant decrease in the activity of CAT, GR. Oral administration of Na₂B₄O₇ at low dose before exposure to K₂Cr₂O₇ is accompanied by an increase in the activity of all studied enzymes and the level of GSH and NPSH, in comparison with the data of animals exposed to Cr⁺⁶. Preventive use of Na₂B₄O₇ in a high dose does not lead to a significant increase in CAT, SOD, GR, and the content of GSH and NPSH, on the contrary, decreases in comparison with the data of rats with chromium-induced brain damage.

Thus, sodium tetraborate in low dose inhibits chromium-induced LPO in the brain, in high, on the contrary, stimulates the FRO lipids. Consequently, tetraborate in a low dose shows an antioxidant effect (neuroprotective effect). It is reported that the toxicity induced by vanadium,⁶¹ titanium,³⁷ aluminum,³⁸ arsenic⁶² can be prevented by the addition of boron compounds. Recently, we have shown that boric acid (5 mg/kg) when combined orally with potassium dichromate (3 mg/kg) inhibits the development of chromium-induced oxidative stress by inhibiting LPO and increasing the power of antioxidant status.⁶³ A prophylactic use of sodium tetraborate at a dose of 4 mg/kg b.w. with chromic intoxication (0.33 LD₅₀) leads to a decrease in MDA level, increased activity of CAT and GSH level, preventing chromium-induced liver damage and genotoxicity of Cr(VI).⁶⁴

Boron (its compounds) under these conditions exhibits an antioxidant property, due to the means to hydroxyl groups⁶⁵ and the ability to form diester bridges between cis-hydroxyl-containing molecules. Another mechanism that reduces the toxicity of chromium to neurons in the brain should be sought, obviously, in the increase in the level of GSH and NPSH and activation of antioxidant enzymes.⁶⁶ The third mechanism is probably related to the fact that sodium tetraborate in low dose has a stabilizing effect affecting the permeability of the HEB under these conditions.³⁹

Thus, the preventive use of Na₂B₄O₇ at a low dose reduces the peroxidation of brain lipids induced by K₂Cr₂O₇ – intoxication, and also increases the activity of antioxidant brain enzymes (SOD, KAT, GR) and increases the capacity of the non-enzyme link (GSH and NPSH) of the antioxidant system. The mechanisms by which sodium tetraborate has a protective effect under conditions of chromium-induced neurotoxicity may be associated with a decrease in the formation of free radicals induced by Cr⁺⁶, an increase in the activity of antioxidant enzymes and a level of antioxidant substances, which in turn neutralize free radicals or through antioxidant effects of direct boron, as a result, the permeability of the blood-brain barrier also decreases.

The use of sodium tetraborate in high dose (72 mg/kg) did not show the expected positive effect; on the contrary, LPO in brain was intensified against a background of low active antioxidant enzymes (KAT, GR) and a low level of GSH and NPSH, which is consistent with the Hu Q. et al (2014), who observed that a low concentration of boron plays a protective role in the development of the spleen, while a high concentration of boron can damage the organs and produce a toxic effect due to the accumulation of MDA and the development of oxidative stress.⁶⁷ Oxidative stress caused by an imbalance between pro- and antioxidant levels⁶³ can initiate several metabolic and functional dysregulation, which ultimately leads to cell death,⁶⁸ i.e. oxidative neuronal necrosis.

It is known that FRO forming upon the reduction of Cr⁺⁶ in Cr⁺³ are mediators of cellular structures.⁶⁹ In this regard, it can be assumed that Na₂B₄O₇ at low doses, increasing antioxidant capacity, preserving the prooxidant-antioxidant status, inhibits LPO in the brain and thereby prevents (destroys) the destabilization and disintegration of neuronal membranes, stabilizes the permeability of HEB, and as a consequence shows neuroprotective effects. However, Na₂B₄O₇ in high dose (72 mg/kg b.w.) reduces the power of antioxidant status, disrupts pro and antioxidant balance, increases LPO, disintegrates and destabilizes membranes, increases the permeability of HEB, thereby increasing the neurotoxicity of K₂Cr₂O₇, i.e. cell membrane in neurons are destroyed and death of brain cells occurs.

Conclusions

A low dose of sodium tetraborate can protect the brain from chromium-induced damage in rats. However, a high dose of Na₂B₄O₇ does not prevent chromium-induced damage in the brain and even increases the neurotoxicity of Cr⁺⁶.

Acknowledgement

The authors are grateful to the anonymous reviewers for their assessment of the study.

Conflict of Interest

The authors declare that they do not have any potential conflicts of interest.

Funding source

To finance the study, the author’s personal funds were used.

References

1. Mahvi A.H. Application of agricultural fibers in pollution removal from aqueous solution. Int. J. Environ. Science Technology. 2008;5(2):275-285.
   CrossRef
2. Mamyrbaev A.A. Toxicology of chromium and its compouds. Aktobe. 2012;284.
3. Papassiopi N, Kontoyianni A, Vaxevanidou K, Xenidis A. Assessment of chromium biostabilization in contaminated soils using standard leaching and sequential extraction techniques. Sci. Total. Environ. 2009;407:925–936.
   CrossRef
4. Becquer T, Quantin C, Sicot M, Boudot J.P. Chromium availability in ultramafic soils from New Caledonia. Science of the Total Environment. 2003;301(1-3):251-261.
   CrossRef
5. Valko M, Morris H and Cronin M.T. Metals, Toxicity and Oxidative Stress. Current Medicinal Chemistry. 2005;12:1161-1208.
   CrossRef
6. Aruldhas M.M, Subramanian S, Sekhar P, Vengatesh G, Chandrahasan G, Govindarajulu P, Akbarsha M.A. Chronic chromium exposure – induced changes in testicular histoarchitecture are associated with oxidative stress: study in non – human primate (Macaca radiata Geoffrey). Human Reprod. 2005;20(10):2801-2813.
   CrossRef
7. Goulart M, Batoreu M.C, Rodrigues A.S, Laires A and Rueff J. Lipoperoxidation products and thiol antioxidants in chromium exposed workers. Mutagenesis. 2005;20:311-315.
   CrossRef
8. Wise S.S, Holmes A.L and Wise J.P. Hexavalent chromium – induced DNA damage and repair mechanisms. Environ. Health. 2008;23(1):39-57.
9. Wang X , Young-Ok S,  Chang Q, Sun L,  Hitron A,  Budhraja A,  Zhang Z,  Ke Z,  Chen F. NADPH Oxidase Activation Is Required in Reactive Oxygen Species Generation and Cell Transformation Induced by Hexavalent Chromium. Toxicol Sci. 2011;123(2):399-410.
   CrossRef
10. Kopec A.K, Kim S, Forgaes A.L, Zacharewsri T.R, Proctor D.M, Harris M.A, Haws L.C, Thompson Ch M. Genome – wide gene expression effects in B6C371 mouse intestinal epithelia following 7 and 90 days of exposure to hexavalent chromium in drinking water. Toxicology and applied pharmacology. 2012;259:13–36.
    CrossRef
11. Casalegno C, Schifanella O, Zennaro E, Marroncelli S, Briant R. Collate literature data on toxicity of Chromium (Cr) and Nickel (Ni) in experimental animals and humans. Supporting publications EN-478 . Available online: www.efsa.europa.eu/publications. 2015;287.
12. Solis-Heredia M.J, Quintanilla-Vega B, Sierra-Santoyo A, Hernandez J.M, Brambila E, Cebrian M.E, Albores A. Chromium increases pancreatic metallothionein in the rat. Toxicology.  2000;142:111–117.
    CrossRef
13. Bagchi D, Balmoori J, Bagchi M, Ye X., Williams C.B, Stohs S.J. Comparative effects of TCDD, endrin, naphthalene and chromium (VI) on oxidative stress and tissue damage in the liver and brain tissues of mice. Toxicology. 2002;175:73-82.
    CrossRef
14. Fatima S, Arivarasu N.A, Banday A.A, Yusufi A.N, Mahmood R. Effect of potassium dichromate on renal brush border membrane enzymes and phosphate transport in rats. Hum Exp. 2005;24(12):631-8.
    CrossRef
15. Soudani N, Troudi A, Amara I.B, Bouaziz H, Boudawara T, Zeghal N. Ameliorating effect of selenium on chromium (VI)-induced oxidative damage in the brain of adult rats. J. Physiol. Biochem. 2012;68:397–409.
    CrossRef
16. Brien T.J, Ceryak S, Patierne S.R. Complexities of chromium carcinogenesis: role of cellular response, repair and recovery mechanism. Mutation Res. 2003;533:3-36. .
    CrossRef
17. Sudha S, Kripa S.K, Shibily P, Shyn J. Elevated Frequencies of Micronuclei and other Nuclear Abnormalities of Chrome Plating Workers Occupationally Exposed to Hexavalent Chromium. Iran J. Cancer. Prev. 2011;3:119-124.
18. Namiesnik J, Rabajiczyk A. Speciation analysis of chromium in enviconmental samples. Critical reviews in environ Sci. Technol. 2012;42(4):327-377.
    CrossRef
19. Fang Z, Zhao M, Zhen H, Chen L, Shi P, Huanol P. Genotoxicity of 3 and hexavalent chromium compounds in vivo and their modes of action on DNA damage in vitro. Plos one 9(8): e103194, doi: 10.1371/ journal Pone. 2014;0103194 .
20. Orellana J.A, Sáez P.J, Shoji K.F, Schalper K.A, Palacios-Prado N, Velarde V, Giaume C, Bennett M.V, Sáez J.C. Modulation of brain hemichannels and gap junction channels by pro-inflammatory agents and their possible role in neurodegeneration. Antioxid Redox Signal. 2009;11(2):369-99.
    CrossRef
21. Suzaki Y, Yoshizumi M, Kagami S, Koyama A, Tsuchiya K, Takeda E, Tamaki T. Hydrogen peroxide stimulates c-Src-mediated big mitogen-activated protein kinase 1 (BMK1) and the MEF2C signaling pathway in PC12 cells: potential role in cell survival following oxidative insults .Biol. Chem. 2002;277:9614-9621.
    CrossRef
22. Bagchi D, Vuchetich P.J, Bagchi M, Hassoun E.A, Tran M.X, Tang L, Stohs S.J. Induction of oxidative stress by chronic administration of sodium dichromate (chromium VI) and cadmium chloride (cadmium II) to rats. Free Radiation Biol Med.  1997;22:471–478.
    CrossRef
23. Travacio M,  María Polo J,  Llesuy S . Chromium(VI) induces oxidative stress in the mouse brain. Toxicology. 2000;150(1-3):137-146.
    CrossRef
24. Iztleuov E.M. Pharmacological correction of reproductive disorders with excess intake of hexavalent chromium (experimental study). Diss. Candidate of Medical Sciences: 00.25-Aktobe. 2007;108.
25. Egemberdieva R.E, Kismanova G.N, Dosmambetova S.K, Darzhanova K.B. Microelement composition and morphological characteristics of the brain of rats when exposed to chemical compounds. Questions of morphology and clinic. – Almaty. 2004;12:412-424.
26. Campbell A. The role of aluminum and copper on neuroinflammation and Alzheimer’s disease.  Journal of Alzheimer’s Disease. 2006;10:165-172.
    CrossRef
27. Nielson F.H. The nutritional importance and pharmacological potential of boron for higher animals and human. In: Goldbach H. E, Rerkasem B, Wimmer M.A, Brown P.H, Thellier M, Bell R.W. (eds) Boron in plant and animal nutrition. Kluwer Academic / Plenum New York. 2002;37-49.
28. Cheng J.Y, Peng K.M, Jin E.H, Zhand Y, Lui Y. Effect of additional boron on tibias of African ostrich chicks. Biol Trace Elem. Res. 2011;144:538-549.
    CrossRef
29. Ying X.Z, Cheng S.W, Wang W, Lin Z.Q, Chen Q.Y, Zhang W, Kuo D.Q, Shen Y, Cheng X.J, Peng L,Luc Z. Effect of boron on osteogenic differentiation of human bone marrow stromal cells. Trace Elem. Res. 2011;144: 306-315.
    CrossRef
30. Turkez H, Geyikoglu F, Tatar A, Keles S, Ozkan A. Effects of some boron compounds on peripheral human blood. Naturforsch. 2007;62:889-896.
    CrossRef
31. Barronco W.T, Kim D.H, Stell S.L, Eckhert C.D. Boric acid inhibits stored Ca+2 release in D-145 prostate cancer cells. Cell Biol. Toxical. 2008;25:309-320.
    CrossRef
32. Korkmaz M,  Uzgören E, Bakirdere S, Aydin F, Ataman O.Y. Effects of dietary boron on cervical cytopathology on micronucleus frequency in exfoliated buccal cells. Environ Toxicol.  2007;22:17-25.
    CrossRef
33. Ornat S.T, Konur M. Cytogenetic Evaluations of Peripheral Blood Samples of Boron Workers. In the proceeding of 2 International Boron Symposium Eshisehir, Turkey.  2004;559-562.
34. Nielsen F.H, Penland J.G.  Boron deprivation alters rat behavior and brain mineral composition differently when fish oil instead of safflower oil is the diet fat source. Nutr Neurosci. 2006;9:105–112.
    CrossRef
35. Barranco W.T, Eckhert C.D. Cellular changes in boric acid-treated DU-145 prostate cancer cells.  J. Cancer. 2006;94:884–890.
    CrossRef
36. Pawa S, Ali S. Boron ameliorates fulminant hepatic failure by counteracting the changes associated with oxidative stress. 2006;160:89-98.
37. Turker H. Effect of boric acid and borax on titanium dioxidate genofoxicity. Appl. Toxicol. 2008;28:654-664.
38. Turkez H, Geyikoglu F. and Cjlak C. The protective effect of boric acid on aluminium – indiced hepatotoxicity and genotoxicity in rats. Turk. J. Biol. 2011;35:293-301.
39. Colak S, Geyikoǧlu F, Keles O.N, Turkez H, Topal A, Unal B. The neuroprotective role of boric acid on aluminum chloride-induced neurotoxicity. Toxicol Ind Health. 2011;27:700–710.
    CrossRef
40. Nielsen F.H. Update on human health effects of boron. Journal of Trace Elem Medicine and  Biology. 2014. http://dx.doi.org/10.1016/j.jtemb.2014.06.023 http://dx.doi.org/10.1016/j.jtemb. 2014.06.023.
    CrossRef
41. Scorei R, Ciubar R, Ciofrangeanu C.M, Mitran V, Cimpean A. Comparative effects of boric acid and calcium tetraborate on breast cancer cells. Biological Trace Element Research. 2008;122:197–205.
    CrossRef
42. Zorzi E.B,  Basso G, Rosolen A.  Bortezomib-mediated 26S proteasome inhibition causes cell-cycle arrest and induces apoptosis in CD-30+ anaplastic large cell lymphoma Leukemia.  2007;21:838-842.
    CrossRef
43. Dhananjayan S, Ismail A, Nawaz Z. Ubiquitin and the control of transcription. Essays in Biochem. 2005;41:69-80.
    CrossRef
44. Navon A. and Ciechanover A. The 26S proteasome: from basic mechanisms to drug targeting. The Journal of Biologocal Chemistry. 2009;284(49):33713-33718.
    CrossRef
45. Meiners S, Ludwig A, Lorenz M, Dreger H, Baumann G, Stangl V, Stangl K. Nontoxic proteasome inhibition activates a protective antioxidant defense response in endothelial cells. Free Radical Biology and Medicine. 2006;40(12):2232-2341.
    CrossRef
46. Fasanaro P, Capogrossi M.C, Martelli F. Regulation of the endothelial cell cycle by the ubiquitin-proteasome system. Cardiovascular Research. 2010;85(2):272-80.
    CrossRef
47. Stangl K, Stangl V. The ubiquitin-proteasome pathway and endothelial (dys)function. Cardiovasc Res. 2010;85(2):281–90.
    CrossRef
48. Iztleuov M.K. Pathogenesis homeostasis disorders caused by excessive intake of chromium in organisms and ways of their correction. Dissertation of the doctor of medical sciences. Moscow.  2004;361.
49. Weir R.J, Fusher R.S. Toxicologic studies on borax and boric acid. Appl. Pharmacol.  1972;23:351-364.
    CrossRef
50. Draper H.H, Hadley M. Malondialdehyde determination as index of lipid peroxidation. Methods Ezymol. 1990;86:421-431.
    CrossRef
51. Koroluk M.A, Ivanova L.I, Mayorova I.G, Tokarev V.E. Method for the determination of catalase activity // Laboratory work. 1988;5:16-18.
52. Ellman G.L. Tissue sulfhydryl groups. Biochem. Biophys. 1959;82:70-77.
    CrossRef
53. Jollow D.J, Mitchell J.R, Zampaglione N, Gillette J.R. Bromobenzene – induced liver necrosis. Protective role of glutathione and evidence for 3,4 – bromobenzene oxide as the hepatotoxic metabolite. Pharmacology.1974; 11:151-169.
    CrossRef
54. Yawata Y, Tanara K.R. Regulatory mechanism of glutathione reductase activity in human red. Biologia. 1974; 43(1):99-109.
55. Chevari S, Andyal T, Shtrenger Ya. Determination of antioxidant blood parameters and their diagnostic value in the elderly // Laboratory work. 1991;10:9-13.
56. Lowry O. H, Rosebrough N. J, Farr A. L and Randall R. J. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 1951;193:265–275.
57. Bielicka A, Bojanowska I, Wisniewshi A. Two Faces of Chromium – Pollutant and Bioelement. Polish Journal of Environmental studies. 2005;14(1):5-10.
58. Bagchi D, Bagchi M, Stohs S.J. Chromium (VI)-induced oxidative stress, apoptotic cell death and modulation of p53 tumor suppressor gene. Mol. Cell. Biochem. 2001;222:149–158.
    CrossRef
59. Iztleuov E.M, Iztleuov M.K, Kydyrbaeva E.A. Prenatal effects of potassium dichromate and their correction in the offspring of rats Medicine and Public Health: materials of the IV International Scientific Conference (Kazan, May M422016) – Kazan: «Young Scientist. 2016;23-26.
60. Hojo Y, Satomi Y. In vitro nephrotoxicity induced in mice by chromium (VI): Involvement of glutathione and chromium (V). Biol Trace Elem Res. 1991;31:21-31.
    CrossRef
61. Geyikoglu F, Turkez H. Boron compounds reduce vanadium tetraoxide genotoxicity in human lymphocytes. Environmental toxicology and pharmacology Q.G. 2008;342-347.
62. Kucukkurt I, Ince S, Demirel H.H, Turkmen K, Arbel E, Cerik Y. The effect of boron on Arsenic – induced lipid peroxidation and antioxidant status in male and femalt rats. Biochem Mol Toxicol. 2015;10:1002./jbt. 21729,
63. Iztleuov M, Umirzakova Z, Iztleuov Ye, Sambaeva S,  Iztleuova G, Yesmukhanova D, Akhmetova A, Medeuova R and Kolishbaeva I. The effect of Chromium and Boron on the Lipid peroxidation and Antioxidant status (in experiment). Biotechnol Ind. J. 2017;13(1):125.
64. Iztleuov Y.M, Kubenova N.N, Ismailova I.V, Iztleuov M.K. Protective Action of Sodium Tetraborate on Chrom-induced Hepato- and Genotoxicity in Rats. Biomedical & Pharmacology Journal. 2017;10(3):1239-1247.
    CrossRef
65. Bolanos L, Lukaszewski K, Bonilla L and Blevins D. Why boron? Physiol. Biochem. 2004;42:907-912.
    CrossRef
66. Ince S, Kucukkurt I, Gigerci I.H, Fidan A.F, Ergavus A. The effects of diatary boric acid and borax supplementation on lipid peroxidation antioxidant activity and DNA damage in rats. of Trace Elements in Melicine and Biology. 2010;24(3):161-164.
    CrossRef
67. Hu Q, Li S, Qiao E, Tang Z, Jin E, Jin G, Gu Y. Effects of Boron on Structure and Antioxidative Activities of Spleen in Rats. Biol Trace Elem Res. 2014;158:73–80.
    CrossRef
68. Subramanian S, Rajendiran G, Sekhar P, Gowri Ch, Govindarajulu A, Aruldhas M.M. Reproductive toxicity of chromium in adult bonnet monkeys (Macaca radiate Geoffrey). Reversible oxidative stress in the semen. Toxicology and Applied pharmacology. 2006;215:237-249.
    CrossRef
69. Pritchard K.A, Ackerman A, Kalyanaraman B. Chromium VI increases endothelial cell expression of ICAM-1 and decreases nitric oxide activity. J. Environ. Pathol. Toxicology Oncology. 2000;19:251-260.