NON ENZYMATIC AND ENZYMATIC ANTIOXIDANT STATUS OF WISTAR RAT ERYTHROCYTE AFTER DIETARY ZINC DEFICIENCY

NON ENZYMATIC AND ENZYMATIC ANTIOXIDANT STATUS OF WISTAR RAT ERYTHROCYTE AFTER DIETARY ZINC DEFICIENCY

Neena Nair ^*, Sunita Bedwal, Pooja Sharma, Aastha Saini, Deepa Kumari and Ranveer Singh Bedwal

Department of Zoology, Cell and Molecular Biology Laboratory, Centre for Advanced Studies University of Rajasthan, Jaipur - 302004, Rajasthan, India.

ABSTRACT: Zinc deficiency is prevalent worldwide. The present study deals with the effect of dietary zinc deficiency on erythrocytes of Wistar rats. Male Wistar rats (40-50 g) were divided into 3 groups: control (ZC) fed 100 μg/g zinc; pair fed (PF) fed 100 μg/g zinc but the diet given was based on the diet consumed the previous day and zinc deficient (ZD) fed 1 μg/g zinc. The experiment was carried out for 2, 4 and 6 weeks. Erythrocyte cell suspension revealed significant increase in catalase activity and decrease in glutathione concentration after zinc deficiency. A significant (P<0.05) increase in erythrocyte lysate superoxide dismutase, glutathione peroxidase and glutathione-s-transferase was observed while glutathione reductase decreased significantly after zinc deficiency. The decreased glutathione reductase coordinates with decreased glutathione concentration making erythrocytes prone to oxidative stress while increased superoxide dismutase, catalase, glutathione peroxidase and glutathione-s-transferase reflects generation of excess free radicals not only due to auto-oxidation of haemoglobin but also due to loss of osmotic fragility leading to oxidative stress indicative of perturbation of erythrocytes on account of dietary zinc deficiency.

Erythrocytes, Zinc deficiency, Catalase, Superoxide dismutase, Glutathione, Glutathione enzymes

INTRODUCTION: Prevalence of nutritional deficiency is predicted to be high in the developing countries ¹ with India being one of them ². Approximately more than half of the world’s population is related to trace element deficiencies which have lead to morbidity and mortality. Mild to moderate zinc deficiency is likely to be prevalent throughout the world although severe zinc deficiency is considered rare ^{3, 4}. It is nutritionally an essential element for human-beings, animals and plants, an absolute requirement in genetic make-up of every cell ⁵.

Reactive oxygen species are continuously being generated within the red blood cells on account of high oxygen tension in arterial blood and haeme Fe²⁺ content. Generation of oxidizing radicals such as 0₂^.-, H₂O₂ and OH^. would lead to oxidative stress⁶. Reactive oxygen species mediated cellular damage effects the membrane integrity accounting for loss of osmotic fragility ⁷ as well as loss of cellular homeostasis with subsequent activation of apoptotic signaling pathway ⁸.

Superoxide dismutase (SOD, EC1.15.1.1) a dimeric metalloenzyme catalyzes the dismutation of superoxide anion ( O₂^.- ) into  molecular  oxygen and hydrogen  peroxide  which  occurs  rapidly (~10⁵ M^-1S^-1 pH 7) ⁹. Correlation between SOD activity and life span of erythrocyte has been deduced by Grzelack et al; ¹⁰ Devi et al., ¹¹ reported an increased Cu-Zn-SOD activity in acute lymphocyte and non lymphocyte leukemia’s which reflects its role in protecting erythrocyte from oxidative stress. Catalase (EC1.11.1.6) a heme protein present in peroxisomes and inner mitochondria converts O₂^.-  which traverses through the membrane into H₂O₂ thereby protecting red blood cells as haemoglobin is a catalyst for peroxidative reactions. Giulivi et al., ¹² reported ~2×10^-10 mol/L H₂O₂ in erythrocytes. Al-Naama et al., ¹³ showed increased antioxidant activities (SOD and CAT) as well as weak correlation with malondialdehyde level involving impairement of antioxidant defense system in erythrocytes of sickle cell anemic patients. GSH a vital antioxidant functions as a radical scavenger involved in reduction of superoxide anion against continuous oxidation of haemoglobin as well as oxidative stress damage ¹⁴ and is responsible for maintaining the red blood cell integrity under certain conditions¹⁵ with concentration of 99% in red blood cell ¹⁶ and 20 µM in plasma ¹⁷.

Glutathione peroxidase (GPx, EC1.11.1.9) catalyses the reduction of hydrogen peroxide and various organic hydroperoxides using GSH protecting not only the cells but also DNA, protein and lipids from oxidative damage ¹⁸. A positive correlation has been reported between SOD in erythrocytes and GPx-1 whole blood ¹⁹ and plasma GPx and free MDA concentration ²⁰. Glutathione reductase (GR, EC 1.6.4.2) catalyses the reduction of glutathione disulphide (GSSG) at the expense of NADPH.H⁺. GR deficiency has been related to several disease states ²¹, reduced erythrocyte life span etc. ^{22, 23} Glutathione-s-transferases (GST, EC 2.5.1.18) catalyzes the nucleophilic addition of GSH to electrophilic centres of a wide variety of potentially toxic compounds protecting the cells from the deleterious effects of reactive oxygen as well as nitrogen species (RONS) and maintaining the cell structure and function ²⁴. The present study evaluates the effect of dietary zinc deficiency on Wistar rat erythrocyte non enzymatic-glutathione and enzymatic-catalase, superoxide dismutase, glutathione peroxidase, glutathione-s-tranferase and glutathione reductase.

MATERIALS AND METHODS:

Synthetic Diet: The synthetic experimental diet was prepared based on ICN Research Diet protocol (1999). The composition of synthetic basal diet: Egg white (180g), corn starch (443g), sucrose (200g), corn oil (100g), cellulose (30g), choline chloride (2 g), A17-76 salt mixture (35 g) , A1N-76 C vitamin antibiotic mix (10 g), DL-methionine (7 g), D-Biotin (20 mg). Zinc contents of the basal diet from each lot was estimated at 213.9 nm in air-acetylene flame on GBC 902 double beam atomic absorption spectrophotometer and zinc concen-tration was adjusted to 1.00 µg/gm and 100 µg/gm by addition of appropriate amounts of zinc sulphate.

Experimental Protocol: Pre-pubertal (30-40 days of age) male Wistar rats (40-50g) were divided into three groups (thirty animals each   for 2, 4 and 6 week) (i) Zinc control (ZC) group fed with diet containing 100 µg/gm zinc (ii) Pair fed (PF) group fed with 100 µg/gm zinc diet but the amount of feed given was equal to the feed consumed by zinc deficient group during the previous day. This group was taken into account so as to study the starvation effects due to reduced intake of diet as well as stress effects. Demineralized water was provided ad libitum (iii) Zinc deficient (ZD) group were fed zinc deficient (1.00 µg/gm) diet and provided demineralized water ad libitum. The animal care and handling was carried out according to the guidelines set by the CPCSEA Committee (No.1678/GO/Re/s/12/CPCSEA dated 16.06.17).

The experiment carried out for 2, 4 and 6 weeks was approved by Departmental Animal Ethics Committee, University of Rajasthan, Jaipur, India. The animals were housed individually in polypropylene cages with stainless steel grills. The polypropylene cages, grills and water bottles were washed with detergent solution, de-mineralized water and finally rinsed in 1% EDTA solution prepared in de-mineralized water so as to avoid contamination and subsequent removal of zinc from cages, grills and bottles. Animals from deficient group were autopsied after 2, 4 and 6 weeks under anaesthesia while supplementation groups were autopsied after 4 weeks. Blood samples from the animals of all the groups were collected by cardiac puncture using heparinized syringe and processed for erythrocyte cell suspension and erythrocyte lysate.

Preparation of Erythrocyte Cell Suspension and Lysate: Red blood cells were washed thrice with PBS (pH 7.4) and erythrocytes were separated from the blood, haemolysed in hypotonic lysing buffer and centrifuged at 27,000 g for 30 min at 4 ºC in Sigma high speed centrifuge using 18015 rotors and membrane was processed by conventional method till free of haemoglobin ²⁵.

Biochemical Estimations: Erythrocyte cell suspension and erythrocyte lysate was obtained using Sigma refrigerated high speed laboratory centrifuge (10,000 rpm). Superoxide dismutase (SOD) ²⁶; catalase ²⁷; glutathione peroxidase (GPx) ^{28, 29}; glutathione-s-transferase (GST) ³⁰ and glutathione reductase (GR) ³¹. Absorbance was read on GBC 911 UV-spectrophotometer, Australia and Systronics UV-vis spectrophotometer 169.

Statistical Analysis: Data are expressed as mean ± SEM. Further, analysis was carried using One Way Analysis of Variance (ANOVA) and if the difference was found significant post-hoc test (Duncan’s Multiple Comparison Test) was carried out and P<0.05 was considered significant. Sigma stat 3.5 software was utilized for analysis (Systat Software Asia Pacific Ltd., Bangalore, India).

RESULTS: Catalase activity in erythrocyte cell suspension increased significantly (P<0.05) after 2, 4 and 6 weeks of zinc deficiency when ZD groups were compared with their respective controls and pairfed groups Table 1.

TABLE 1: CATALASE ACTIVITY AND GLUTATHIONE CONCENTRATION IN WISTAR RAT ERYTHROCYTE CELL SUSPENSION AFTER 2, 4 AND 6 WEEKS OF ZINC DEFICIENCY. (MEAN ± SEM)

ZC vs. PF = a, PF vs. ZD = b, ZC vs. ZD = c. Significant (P<0.05) = *. Multiple comparison of means were performed separately for 2, 4 and 6 weeks sub groups.

Decreased (P<0.05) GSH concentration was evident after 2, 4 and 6 weeks of zinc deficiency when groups were compared Table 1. Superoxide dismutase activity in erythrocyte lysate increased significantly after 2, 4 and 6 weeks of zinc deficiency when ZD groups were compared with their respective controls and paired groups Table 2. Significant (P<0.05) increase was observed in GPx and GST activities after zinc deficiency Table 2. Decrease was significant (P<0.05) in erythrocyte lysate GR activity after 2, 4 and 6 week of zinc deficiency when ZD groups were compared with their respective control groups as well as PF groups Table 2.

TABLE 2: BIOCHEMISTRY OF WISTAR ERYTHROCYTE CELL LYSATE AFTER 2, 4 AND 6 WEEKS OF ZINC DEFICIENCY. (MEAN ± SEM)

ZC vs. PF = a, PF vs. ZD = b, ZC vs. ZD = c. Significant (P<0.05) = *. Multiple comparison of means were performed separately for 2, 4 and 6 weeks sub groups.

DISCUSSION: Erythrocytes are sensitive to reactive oxygen species (ROS) generated constantly even during normal physiological events as auto-oxidation leads to generation of superoxide³². Although ROS are removed by antioxidant defense mechanism but an imbalance may lead to modification either to the membrane or macro-molecules ^{14, 24, 33}. Superoxide dismutase is a major antioxidant enzyme in erythrocyte where O₂^•- are continuously generated by auto-oxidation of haemoglobin. Zinc deficiency in the present study enhanced SOD activity which can be correlated with increased resistance towards toxicity/ damaging effects of superoxide radical. Cooperative action of O₂^•- and H₂O₂ would generate a potent oxidant OH^• involving reduction of Fe III to Fe II ³⁴ leading to enhanced osmotic fragility ⁷ and ultimately cellular injury. Contrary to our observation decrease in erythrocyte SOD activity in 4 week old Sprague-Dawley rats fed zinc deficient diet for 10-16 days was reported ³⁵ whereas Song et al., ³⁶ could not record any variation in zinc deficient rats compared to zinc adequate / control rats. Since H₂O₂ (generated from O₂^•- by SOD) serves as a substrate for Fenton reaction to generate highly reactive OH^•, catalase and glutathione peroxidase enzymes functions as ROS scavenging enzymes by limiting the accumulation of H₂O₂ or organic peroxides ^{37, 38}.

The concentration of glutathione decreased significantly in the present study. Cells deprived of GSH are more prone to oxidative stress even under normal circumstances, since the ability to scavenge or detoxify ROS/FOR is impaired ³⁹ and depletion of GSH levels in erythrocytes has been associated with oxidant burden ⁴⁰. Enhanced oxygen free radical is associated with decreased plasma GSH: GSSG ratio which may be one of the factors responsible for disease states in humans ⁴¹. Decreased food intake, intermittent diarrhea ^{42, 43} leading to decreased protein concentration in erythrocytes of zinc deficient and pair fed groups might be one of the reasons for decreased GSH. Furthermore, oxidation of GSH/GSSG by GPx and GST for scavenging ROS indicative of intracellular redox imbalance has also been correlated to its decreased concentration ⁴⁴. Decrease in GSH may probably be due to decrease in its formation as one of its requirements is GR which has also decreased in the present study. Depletion of GSH may have adverse consequences as it weakens the defense against oxidative stress which may have deleterious effect leading to dysfunction of the cell.

Glutathione peroxidase-1 (GPx-1) is found exclusively in erythrocytes while GPx-3 is a seleno-protein ^{45, 46} with membrane stability (cellular as well as subcellular) dependent on this enzyme. Knock out experiments with catalase or GPx-1 genes in mouse showed that GPx-1 reduces organic peroxides ⁴⁷. The present study demonstrates increased GPx activities after zinc deficiency. Increased GPx activity has been reported in erythrocyte after zinc deficiency ⁴⁸ and after transport stress (GPx-1; beef-cattle Sahin et al., ⁴⁹). Similarly, increased GPx-1 activities have been reported from acute lymphocytic and non lymphocytic leukemia ¹¹. GPx-4 reduces the phospholipid hydroperoxides in normal red cell/mammalian cells ⁵⁰ but is dependent on glutathione. Deficiency would potentiate oxidative damage to the erythrocytes. Thus, the two FOR / ROS scavenging enzymes-catalase and GPx responsible for eliminating H₂O₂/organic peroxides probably function in a coordinated manner to overcome the oxidative stress in erythrocytes after zinc deficiency. Contrary to this, Muller et al., ⁵¹ reported that H₂O₂ generated in human erythrocytes are removed only by catalase.

Glutathione reductase and glutathione-s-transferase decreased in a duration dependent manner after dietary zinc deficiency in the present study. The decreased glutathione-s-transferase activity can be correlated to decreased consumption of GST cofactor-GSH which can account for depletion of intracellular GSH with resultant decreasing levels of relative Erythrocyte-GST activity. Authors^{52, 53} have reported inactivation of erythrocyte GST on account of oxidative stress. The decrease in erythrocyte glutathione reductase has been reported when zinc deficiency was compared with the control ⁴⁸. Glutathione reductase has a significant role in protecting haemoglobin, erythrocyte enzymes and membrane from oxidative damage by increasing the level of GSH ⁵⁴. The decreased GR activity coordinated well with reduced level of GSH in the present study which might have led to increased oxidative stress to erythrocytes of dietary zinc deficiency and increased concentration of GSSG.

CONCLUSION: Dietary zinc deficiency for a period of 2, 4 and 6 weeks potentiated non enzymatic and enzymatic antioxidants of erythrocytes which may be one of the causative factors for dysfunction. If oxidant molecules are not denatured by antioxidant mechanism then erythrocyte destruction would occur. This is indicative that microelement zinc in diet is crucial for the normal function of mammalian erythrocytes.

ACKNOWLEDGEMENT: Dr. Pooja Sharma thanks Department of Science & Technology, New Delhi for the award of Inspire fellowship (No. DST/INSPIRE Fellowship/2011/[102]). Ms. Aastha Saini thanks University Grants Commission for the award of Junior Research Fellowship (No. 22/06/2014 (1) EU-V) and Senior Fellowship. Dr. Deepa Kumari thanks University Grants Commission for award of post doctoral fellowship ([No. F.15-1/2011-12/PDFWM-2011-12-GE-RAJ-3826 (SA-II)]).

The authors thank Head, Department of Zoology, University of Rajasthan and Centre for Advanced Studies programme for providing necessary facilities.

CONFLICT OF INTEREST: There is no conflict of interest.

REFERENCES:

1. Bailey RL, West Jr KP and Black RE: The epidemiology of global micronutrient deficiencies. Annals of Nutrition & Metabolism 2015; 66(S-2): 22-33.
2. Bharti HP, Kavthekar SO, Kavthekar SS and Kurane AB: Prevalence of micronutrient deficiencies clinically in rural school going children. International Jour of Contemporary Pediatrics 2018; 5(1): 234-238.
3. Hambidge KM: Micronutrient bioavailability: Dietary reference intakes and a future perspective. American Journal of Clinical Nutrition 2010; 9: 1430-1432.
4. Nagata M, Kayanoma M, Takahashi T, Kaneko T and Hara H: Marginal zinc deficiency in pregnant rats impairs bone matrix formation and bone mineralization in their neonates. Biological Trace Element Research 2010; 142(2): 190-199.
5. Prasad AS: Zinc: A miracle element. Its discovery and impact on human health. JSM Clinical Oncology Research 2014; 2(4): 1030-1037.
6. Nimse SB and Pai D: Free radicals, natural antioxidants and their reaction mechanisms. Royal Society of Chemistry Advances 2015; 5: 27986-28006.
7. Sharma P, Nair N, Kumari D and Bedwal RS: Morphology and osmotic fragility in Wistar rat erythrocytes after zinc deficiency and supplementation. Indian Journal of Applied Research 2016; 6(10): 314-318.
8. Redza-Dutordoir M and Averill-Bates DA: Activation of apoptosis signalling pathways by reactive oxygen species. Biochimica et Biophysica Acta 2016; 1863: 2977-2992.
9. Samuel ELG, Marcano D, Berka V, Bitner BR, Wu G, Potter A, Fabian RH, Pautler RG, Kent TA, Tsai AL and Tour JM: Highly efficient conversion of superoxide to oxygen using hydrophilic carbon clusters. Proceedings of National Academy of Sciences, USA 2015; 112(8): 2343-2348.
10. Grzelack A, Kruszewski M, Macierzyńsk E, Piotrowski L, Pułaski L, Rychlik B and Bartosz G: The effect of superoxide dismutase knockout on the oxidative stress parameters and survival of mouse erythrocytes. Cellular & Molecular Biology Letters 2008; 14: 23-34.
11. Devi GS, Prasad MH, Saraswathi I, Raghu D, Rao DN and Reddy PP: Free radicals, antioxidant enzymes and lipid peroxidation in different types of leukemia’s. Clinica Chimica Acta 2000; 293: 53-62.
12. Giulivi C, Hochstein P and Davies KJ: Hydrogen peroxide production by red blood cells. Free Radical Biology & Medicine 1994; 16: 123-129.
13. Al-Naama LM, Hassan MK and Mehdi JK: Association of erythrocytes antioxidant enzymes and their cofactors with markers of oxidative stress in patients with sickle cell anemia. Qatar Medical Journal 2015; 2: 14. https://doi.org/ 10.5339/qmj.2015.14.
14. Halliwell B and Gutteridge JM: Free radicals in biology and medicine. Oxford Press New York, Edition 5^th. 2015.
15. Raftos JE, Whilier S and Kuchel PW: Glutathione synthesis and turnover in human erythrocyte: Alignment of a model based on detailed enzyme kinetics with experimental data. Journal of Biological Chemistry 2010; 285: 23557-23567.
16. Marí M, Morales A, Colell A, García-Ruiz C and  Fernández-Checa JC: Mitochondrial glutathione, a key survival antioxidant. Antioxid Redox Signal 2009; 11(11): 2685-700
17. Lushchak VI: Glutathione homeostasis and functions: potential targets for medical interventions. Journal of Amino Acids 2012; 2012: 736837.
18. Ighodaro OM and Akinloye OA: First line defence antioxidants-superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX): Their fundamental role in the entire antioxidant defence grid. Alexandria Journal of Medicine 2017. https://doi.org/10.1016/j.ajme.2017. 09.001.
19. Covas MI, Coca L, Ricos C and Marrugat J: Biological variation of superoxide dismutase in erythrocytes and glutathione peroxidase in whole blood. Clinical Chemistry 1997; 43: 1991-1993.
20. Cilary PH, Bratter P, Negretti de Bratter, Khassanava L and Etrenne JC: Metal ion in biology and medicine. John Libbey Eurotext, Paris 1998; 5: 400-404.
21. Zwieten R van, Verhoeven, AJ and Roos D: Inborn defects in the antioxidant systems of human red blood cells. Free Radical Biology Medicine 2014; 67: 377-386.
22. Kmerbeek NM, van Zweieten R, deBoer M, Morren G, Vuil H, Bannink N, Lincke C, Dolman KM, Becker K, Schirmer RH, Gromer S and Roos D: Molecular basis of glutathione reductase deficiency in human blood cells. Blood 2007; 109: 3560-3566.
23. Qiao M, Kisgati M, Cholewa JM, Zhu W, Smart EJ, Sulistio MS and Asmis R: Increased expression of glutathione reductase in macrophages decreases atherosclerosis lesion formation in low density lipoprotein receptor-deficient mice. Arteriosclerosis Thrombosis and Vascular Biology 2007; 27: 1375-1382.
24. Espinosa-Diez  C, Miguel  V, Mennerich D, Kietzmann T, Sánchez-Pérez  P,  Cadenas S  and Lamas S:  Antioxidant responses and cellular adjustments to oxidative stress. Redox Biology 2015; 6: 183-197.
25. Steck TL and Kant JA: Preparation of impermeable ghosts and inside-out vesicles from human erythrocyte membranes. Methods in Enzymology 1974; 31: 172-180.
26. Kono Y: Generation of superoxide radical during auto-oxidation of hydroxylamine and an assay for superoxide dismutase. Archives of Biochemistry and Biophysics 1978; 186: 189-195.
27. Sinha AK: Colorimetric assay of catalase. Analytical Biochemistry 1972; 47: 389-394.
28. Paglia DE and Valentine WN: Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. Journal of Laboratory Clinical Metabolism 1967; 70: 158-169.
29. Briviba K, Kissner R, Koppenol WH and Sies H: Kinetic study of the reaction of glutathione peroxidase with peroxynitrite. Chemical Research in Toxicology 1998; 11: 1398-1401.
30. Warholm M, Guthenberg C, Von Bohr C and Mannervik B: Glutathione transferase from human liver. Methods in Enzymology 1985; 113: 499-505.
31. Carlberg I and Mannervik B: Glutathione reductase. Methods in Enzymology 1985; 113: 484-490.
32. Mohanty JG, Nagababu E and Rifkind JM: Red blood cell oxidative stress impairs oxygen delivery and induces red blood cell aging. Frontiers in Physiology 2014. doi: 10.3389/fphys.2014.00084.
33. Rahal A, Kumar IA, Singh V, Yadav B, Tiwari R, Chakraborty S and Dhama K: Oxidative stress, pro-oxidants, and antioxidants: The Interplay. Bio Med Res Inter 2014; doi.org/10.1155/2014/761264.
34. Liochev SI and Fridovich I: The Haber-Weiss cycle-70 years later: An alternative view. Redox Report 2002; 7(1): 55-57.
35. Taysi S, Cikman O, Kaya A, Demircan RB, Gumustekin K, Yilmaz A, Boyuk A, Keles M, Akyuz M and Turkeli M: Increased oxidant stress and decreased antioxidant status in erythrocytes of rats fed with zinc-deficient diet. Biological Trace Element Research 2008; 123: 161-167.
36. Song Y, Leonard SW, Traber MG and Ho E: Zinc deficiency affects DNA damage, oxidative stress, anti-oxidant defenses and DNA repair in rats. Journal of Nutrition 2009; 139: 1626-1631.
37. Covas MI, Coca L, Ricos C and Marrugat J: Biological variation of superoxide dismutase in erythrocytes and glutathione peroxidase in whole blood. Clinical Chemistry 1997; 43: 1991-1993.
38. Ho Y-S, Xiong Y, Ma W, Spector A and Ho DS: Mice lacking catalase develop normally but show differential sensitivity to oxidant tissue injury. Journal of Biological Chemistry 2004; 279: 32804-32812.
39. Kurutas EB: The importance of antioxidants which play the role in cellular response against oxidative/nitrosative stress: Current state. Kurutas Nutrition J 2016; 15: 71-92.
40. Rao GM, Naser SA and Vandana: Reduced blood glutathione and erythrocyte stability in osteoarthritis. Biomedical Research 2005; 16(3): 201-203.
41. Zitka O, Skalickova S, Gumulec J, Masarik M, Adam V, Hubalek J, Trnkova1 L, Kruseova J, Eckschlager T and Kizek R: Redox status expressed as GSH: GSSG ratio as a marker for oxidative stress in paediatric tumour patients. Oncology Letters 2012; 4: 1247-1253.
42. Kumari D, Nair N and Bedwal RS: Effect of dietary zinc deficiency on testes of Wistar rats: Morphometric and cell quantification studies. Journal of Trace Element Medicine and Biology 2011; 25: 47-53.
43. Joshi S, Nair N and Bedwal RS: Dietary zinc deficiency affects dorso-lateral and ventral prostate of Wistar rats: Histological, biochemical and trace element study. Biological Trace Element Research 2014; 161(1): 91-100.
44. Circu ML and Aw TY: Glutathione and modulation of cell apoptosis. Biochimica et Biophysica Acta 2012; 1823: 1767-1777.
45. Avissar N, Within JC, Allen PZ, Palmer IS and Cohen HJ: Anti-human plasma glutathione peroxidase antibodies. Immunologic investigations to determine plasma glutathione peroxidase protein and selenium content in plasma. Blood 1989; 73: 318-323.
46. Collins JF: Molecular, genetic, and nutritional aspects of major and trace minerals. Academic Press, New York, London 2017: 449-456.
47. Johnson RM, Ho YS, Yu DY, Kuypers FA, Ravindernath Y and Goyette Jr GW: The effects of disruption of genes for peroxiredoxin-2, glutathione peroxidase-1 and catalase from erythrocyte oxidative metabolism. Free Radical Biology and Medicine 2010; 48: 519-525.
48. Taysi S, Cikman O, Kaya A, Demircan B, Gumustekin K, Yilmaz A, Boyak A, Keles M, Akyuz M and Turkeb M: Increased oxidant stress and decreased antioxidant status in erythrocytes of rats fed with zinc deficient diet. Biological Trace Element Research 2008; 123: 161-167.
49. Sahin T, Gurgoze SY, Camkerten I and Yuksek N: The effect of transport stress on erythrocytes glutathione peroxidase activity, plasma concentration of lipid peroxidation and some trace element levels in beef-cattle Journal of Animal and Veterinary Advances 2009; 8: 222-227.
50. Imai H and Nakagawa Y: Biological significance of phospholipid hydroperoxide glutathione peroxidase (PHGPx, GPx 4) in mammalian cells. Free Radical Biology and Medicine 2003; 34(2): 145-169.
51. Muller S, Hans-Dieter R and Stremmel W: Direct evidence for catalase as predominant H₂O₂-removing enzyme in human erythrocytes. Blood 1997; 90(12): 4973-4978.
52. Evelo CTA: Toxicological stress indicators in human red blood cells; changes in glutathione and glutathione-s-transferase as biological markers for electrophilic and oxidative stress. Thesis, Maastricht 1995; 77-100.
53. Spooren AAMG and Evelo CTA: In-vitro haematotoxic effects of three methylated hydroxylamines. Archives of Toxicology 1997; 71: 299-305.
54. Chang JC, Vander Hoeven LH and Haddox CH: Glutathione reductase in the red blood cells. Annals of Clinical and Laboratory Science 1978; 8: 23-29.
    

    ---

    How to cite this article:

    Nair N, Bedwal S, Sharma P, Saini A, Kumari D and Bedwal RS: Non enzymatic and enzymatic antioxidant status of Wistar rat erythrocyte after dietary zinc deficiency. Int J Pharm Sci & Res 2019; 10(2): 763-68. doi: 10.13040/IJPSR.0975-8232.10(2).763-68.

    ---

    All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.

    ---