Bìol. Tvarin. 2020; 22 (2): 38–42.
Received 01.05.2020 ▪ Accepted 01.06.2020 ▪ Published online 01.07.2020

The corrective effect of chromium and zinc citrates on NO-synthase activity of erythrocytes in rats with streptozotocin diabetes

O. Slivinska1, R. Iskra2

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1Andrei Krupynskyi Lviv Medical Academy,
70 Doroshenko str., Lviv, 79007, Ukraine

2Institute of Animal Biology NAAS,
38 V. Stus str., Lviv, 79034, Ukraine

The aim of the research was to investigate the effect of chromium and zinc citrates on the NO-synthase (NOS) activity of erythrocytes in rats with streptozotocin diabetes. In three series of investigations rats were divided into four groups (I — control, II, III and IV — experimental) each one containing 7 animals. During four weeks, in the first series of investigations chromium citrate was added with water to the main diet of animals in III and IV groups in amounts of 10 and 25 μg Cr3+/kg of body weight; in the second series, zinc citrate was added in amounts of 20 and 50 mg Zn2+/kg of body weight respectively. In the third series, the animals of III group received chromium citrate in amount 25 μg Cr3+/kg of body weight and zinc citrate in amount 50 mg Zn2+/kg of body weight with water. The rats of II experimental group in all series received clean water with no citrates added. A month later, in animals of all experimental groups on the background of a 24-hour fasting an experimental diabetes mellitus was induced by a single intraperitoneal injection of streptozotocin in amounts of 45 mg/kg of body weight with the previous injection of nicotinamide. Diabetes occurred on the third day. Animals with a glucose concentration of 14 mmol/L were used for the experiment. The material for the investigation was the blood of rats, in which the concentration of glucose and the relative count of glycosylated hemoglobin were determined, in erythrocytes — NO-synthase activity: general, inducible and constitutive. As a result of the conducted research, it has been found that under streptozotocin induced experimental diabetes in erythrocytes of rats of experimental group II in both series of investigations the activity of general and inducible NOS increased, while the activity of the constitutive NOS did not change compared with the animals of the control group. The introduction of chromium and zinc citrates into the animals’ diet in the above mentioned doses led to the decrease in the activity of the general and inducible NOS compared with the animals of group II with streptozotocin diabetes, indicating a positive effect of the studied microelements on NOS activity in erythrocytes of rats. Thus, the use of chromium and zinc citrates in the diet of rats with diabetes has a normalizing effect on the state of NOS activity, what can reduce the harmful influence of hyperglycemia on the development of oxidative and nitrosative stress.

Keywords: rats, diabetes mellitus, citrates, chromium, zinc, erythrocytes, nitrogen oxide, NO synthase

  1. Bardsley JK, Want LL. Overview of diabetes. Critical Care Nursing Quarterly. 2004; 27 (2): 106‒112. https://doi.org/10.1097/00002727-200404000-00002
  2. Bharti SK, Singh SK. Metal based drugs: current use and future potential. Der Pharmacia Lettre. 2009; 1 (2): 39‒51. Available at: https://www.researchgate.net/publication/267961271
  3. Bhattacharya S, Chakraborty Patra S, Basu Roy S, Kahn NN, Sinha AK. Purification and properties of insulin-activated nitric oxide synthase from human erythrocyte membranes. Arch. Physiol. Biochem. 2001; 109 (5): 441–449. https://doi.org/10.1076/apab.109.5.441.8042
  4. Birgani GA, Ahangarpour A, Khorsandi L, Moghaddam HF. Anti-diabetic effect of betulinic acid on streptozotocinnicotinamide induced diabetic male mouse model. Braz. J. Pharm. Sci. 2018; 54 (2): e17171. https://doi.org/10.1590/s2175-97902018000217171
  5. Cockcroft JR, Webb DJ, Wilkinson IB. Arterial stiffness, hypertension and diabetes mellitus. J. Hum. Hypertens. 2000; 14: 377‒380. https://doi.org/10.1038/sj.jhh.1001023
  6. Dejam A, Hunter CJ, Pelletier MM, Hsu LL, Machado RF, Shiva S, Power GG, Kelm M, Gladwin MT, Schechter AN. Erythrocytes are the major intravascular storage sites of nitrite in human blood. Blood. 2005; 106 (2): 734‒739. https://doi.org/10.1182/blood-2005-02-0567
  7. Doddigarla Z, Parwez I, Abidi S, Ahmad J. Effect of chromium picolinate and melatonin either in single or in a combination in alloxan induced male Wistar rats. J. Biomedical Sci. 2016; 6: 1. https://doi.org/10.4172/2254-609X.100051
  8. Drel VR. The main mechanisms of the onset and development of diabetic complications: the role of nitric stress. Biol. Stud. 2010; 4 (2): 141–158. https://doi.org/10.30970/sbi.0402.085
  9. Dunn MF. Zinc-ligand interactions modulate assembly and stability of the insulin hexamer — a review. Biometals. 2005; 18: 295‒303. https://doi.org/10.1007/s10534-005-3685-y
  10. Hadwan MH, Almashhedy LA, Alsalman ARS. Study of the effects of oral zinc supplementation on peroxynitrite levels, arginase activity and NO synthase activity in seminal plasma of Iraqi asthenospermic patients. Reprod. Biol. Endocrinol. 2014; 12: 1. https://doi.org/10.1186/1477-7827-12-1
  11. Heltianu C, Guja C. Role of nitric oxide synthase family in diabetic neuropathy. J. Diabetes Metab. 2011; S5: 002. https://doi.org/10.4172/2155-6156.S5-002
  12. Higaki Y, Hirshman MF, Fujii N, Goodyear LJ. Nitric oxide increases glucose uptake through a mechanism that is distinct from the insulin and contraction pathways in rat skeletal muscle. Diabetes. 2001; 50 (2): 241–247. https://doi.org/10.2337/diabetes.50.2.241
  13. James PE, Lang D, Tufnell-Barret T, Milsom AB, Frenneaux MP. Vasorelaxation by red blood cells and impairment in diabetes. Circ. Res. 2004; 94 (7): 976–983. https://doi.org/10.1161/01.RES.0000122044.21787.01
  14. Kahn NN, Acharya K, Bhattacharya S, Acharya R, Mazumder S, Bauman WA, Sinha AK. Nitric oxide: the “second messenger” of insulin. IUBMB Life. 2000; 49 (5): 441–450. https://doi.org/10.1080/152165400410308
  15. Kleinbongard P, Schulz R, Rassaf T, Lauer T, Dejam A, Jax T, Kumara I, Gharini P, Kabanova S, Özüyaman B, Schnürch HG, Gödecke A, Weber AA, Robenek M, Robenek H, Bloch W, Rösen P, Kelm M. Red blood cells express a functional endothelial nitric oxide synthase. Blood. 2006; 107 (7): 2943–2951. https://doi.org/10.1182/blood-2005-10-3992
  16. Kopilas MA, Dang LNT, Anderson HDI. Effect of dietary chromium on resistance artery function and nitric oxide signaling in the sucrose-fed spontaneously hypertensive rat. J. Vasc. Res. 2007; 44 (2): 110–118. https://doi.org/10.1159/000098483
  17. Kosiakova GV, Gulaya NN. The N-stearoylethanolamine effect on the NO-synthase way of nitrogen oxide formation and phospholipid composition of erythrocyte membranes in rats with streptozotocine diabetes. Ukr. Biochem. J., 2007; 79 (6): 53–59. Available at: http://ubj.biochemistry.org.ua/index.php/journal-archiveac/2007qq/6-gfs/4428-n-no (in Ukrainian)
  18. Li H, Raman CS, Glaser CB, Blasko E, Young TA, Parkinson JF, Whitlow M, Poulos TL. Crystal structures of zinc-free and -bound heme domain of human inducible nitric-oxide synthase. Implications for dimer stability and comparison with endothelial nitric-oxide synthase. J. Biol. Chem. 1999; 274: 21276–21284. https://doi.org/10.1074/jbc.274.30.21276
  19. Mitchell DA, Erwin PA, Michel T, Marletta MA. S-nitrosation and regulation of inducible nitric oxide synthase. Biochemistry. 2005; 44 (12): 4636–4647. https://doi.org/10.1021/bi0474463
  20. Niedowicz DM, Daleke D. The role of oxidative stress in diabetic complications. Cell Biochem. Biophys. 2005; 43: 289–330. https://doi.org/10.1385/CBB:43:2:289
  21. Panchyshyn OB, Panasyuk NB, Biletska LP, Sklyarov AY. The changes of NO-synthases and arginase activity in pancreatic tissue under conditions of L-arginine and aminoguanidine administration in streptozocin-induced hyperglycemia. Tavriya Medical and Biological Messenger. 2012; 15 (3/1 (59)): 260–262. Available at: http://dspace.nbuv.gov.ua/handle/123456789/44702 (in Ukrainian)
  22. Pari L, Uma Maheswari J. Hypo-glycemic effect of Musa sapientum L. in alloxan-induced diabetic rats. J. Ethnopharmacol. 1999; 68 (3): 321–325. https://doi.org/10.1016/S0378-8741(99)00088-4
  23. Seven A, Güzel S, Seymen O, Civelek S, Bolayirli M, Yigit G, Burçak G. Nitric oxide synthase inhibition by L-NAME in streptozotocin induced diabetic rats: impacts on oxidative stress. Tohoku J. Exp. Med. 2003; 199 (4): 205–210. https://doi.org/10.1620/tjem.199.205
  24. Shrivastava R, Upreti RK, Seth PK, Chaturvedi UC. Effects of Chromium on the immune system. FEMS Immunol. Med. Microbiol. 2002; 34 (1): 1–7. https://doi.org/10.1111/j.1574-695X.2002.tb00596.x
  25. Stuehr DJ, Griffith OW. Mammalian nitric oxide synthases. In: Meister A (ed.). Advances in Enzymology and Related Areas of Molecular Biology. 1992; 65: 287–346. https://doi.org/10.1002/9780470123119.ch8
  26. Sumbaev VV, Yasynskaya IN. The effect of DDT on the nitric oxide synthase activity in the liver, lungs and brain of rats. Modern Problems of Toxicology. 2000; 3: 3–7. Available at: http://medved.kiev.ua/arhiv_mg/st_2000/00_3_1.htm (in Russian)
  27. Szabó C, Ischiropoulos H, Radi R. Peroxynitrite: biochemistry, pathophysiology and development of therapeutics. Nat. Rev. Drug. Discov. 2007; 6: 662–680. https://doi.org/10.1038/nrd2222
  28. Tian L, Lawrence DA. Metal-induced modulation of nitric oxide production in vitro by murine macrophages: lead, nickel, and cobalt utilize different mechanisms. Toxicol. Appl. Pharmacol. 1996; 141 (2): 540–547. https://doi.org/10.1006/taap.1996.0320
  29. Van den Oever I, Raterman HG, Nurmohamed MT, Simsek S. Endothelial dysfunction, inflammation, and apoptosis in diabetes mellitus. Mediators of Inflammation. 2010: 792393. https://doi.org/10.1155/2010/792393
  30. Vitak TY, Wasser SP, Nevo E, Sybirna NO. Effect of medicinal mushrooms on L-arginine/NO system in red blood cells of streptozotocin-induced diabetic rats. Advances in Diabetes and Metabolism. 2016; 4 (2): 25–31. https://doi.org/10.13189/adm.2016.040201 Available at: http://www.hrpub.org/journals/article_info.php?aid=3642
  31. Wolff SP, Jiang ZY, Hunt JV. Protein glycation and oxidative stress in diabetes mellitus and ageing. Free Radic. Biol. Med. 1991; 10 (5): 339–352. https://doi.org/10.1016/0891-5849(91)90040-A
  32. Yamamoto A, Wada O, Manabe S. Evidence that chromium is an essential factor for biological activity of low-molecular weight chromium-binding substance. Biochem. Biophys. Res. Commun. 1989; 163 (1): 189–193. https://doi.org/10.1016/0006-291X(89)92119-0
  33. Yamaoka J, Kume T, Akaike A, Miyachi Y. Suppressive effect of zinc ion on iNOS expression induced by interferon-γ or tumor necrosis factor-α in murine keratinocytes. J. Dermatol. Sci. 2000; 23 (1): 27–35. https://doi.org/10.1016/S0923-1811(99)00062-6




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