Borate Ameliorates Sodium Nitrite-Induced Oxidative Stress Through Regulation of Oxidant/Antioxidant Status: Involvement of the Nrf2/HO-1 and NF-κB Pathways


The widespread industrial use of nitrite in preservatives, colorants, and manufacturing rubber products and dyes increases the possibilities of organ toxicity. Lithium borate (LB) is known as an antioxidant and an oxidative stress reliever. Therefore, this study is aimed at examining the effect of LB on nitrite-induced hepatorenal dysfunction. Twenty-eight male Swiss mice were divided into four equal groups. Group 1, the control group, received saline. Group 2 received LB orally for 5 consecutive days at a dose of 15 mg/kg bw. Group 3, the nitrite group, received sodium nitrite (NaNO2) on Day 5 (60 mg/kg bw intraperitoneally). Group 4, the protective group (LB + NaNO2 group), received LB for 5 days and then a single dose of NaNO2 intraperitoneally on Day 5, the same as in Groups 2 and 3, respectively. Samples of blood and kidney were taken for serum analysis of hepatorenal biomarkers, levels of antioxidants and cytokines, and the expression of genes associated with oxidative stress and inflammation. NaNO2 intoxication increased markers of liver and kidney functions yet decreased reduced glutathione (GSH), superoxide dismutase (SOD), and catalase activities in blood. NaNO2 also increased the expression of tumor necrosis factor (TNF-α), interleukin-1β and interleukin-6 (IL-1β and IL-6). Pre-administration of LB protected mice from oxidative stress, lipid peroxidation, and the decrease in antioxidant enzyme activity. Moreover, LB protected mice from cytokine changes, which remained within normal levels. LB ameliorated the changes induced by NaNO2 on the mRNA of nuclear factor erythroid 2-related factor 2 (Nfr2), heme oxygenase-1 (HO-1), nuclear factor-kappa B (NF-κB), transforming growth factor-beta 2 (TGF-β2), and glutathione-S-transferase (GST) as determined using quantitative real-time PCR (qRT-PCR). These results collectively demonstrate that LB ameliorated NaNO2-induced oxidative stress by controlling the oxidative stress biomarkers and the oxidant/antioxidant state through the involvement of the Nrf2/HO-1 and NF-κB signaling pathways.

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Data Availability

Data are available from the corresponding author upon request.


  1. 1.

    Adelakun SA, Ukwenya VO, Ogunlade BS, Aniah AJ, Ibiayo GA (2019) Nitrite-induced testicular toxicity in rats: therapeutic potential of walnut oil. JBRA Assist Reprod 23(1):15–23.

    Article  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Dennis M, Wilson L (2003) Nitrates and nitrites. In: Encyclopedia of Food Sciences and Nutrition. Reference Work 2nd ed.

  3. 3.

    Kostić T, Andrić S, Kovacević R, Marić D (1998) The involvement of nitric oxide in stress-impaired testicular steroidogenesis. Eur J Pharmacol 346(2-3):267–273.

    Article  PubMed  Google Scholar 

  4. 4.

    Lundberg JO, Weitzberg E, Gladwin MT (2008) The nitrate-nitrite-nitric oxide pathway in physiology and therapeutics. Nat Rev Drug Discov 7(2):156–167.

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Shiva S (2013) Nitrite: a physiological store of nitric oxide and modulator of mitochondrial function. Redox Biol 1(1):40–44.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Carlstrom M, Montenegro MF (2019) Therapeutic value of stimulating the nitrate-nitrite-nitric oxide pathway to attenuate oxidative stress and restore nitric oxide bioavailability in cardiorenal disease. J Intern Med 285(1):2–18.

    CAS  Article  PubMed  Google Scholar 

  7. 7.

    Carlström M, Persson AE, Larsson E, Hezel M, Scheffer PG, Teerlink T, Weitzberg E, Lundberg JO (2011) Dietary nitrate attenuates oxidative stress, prevents cardiac and renal injuries, and reduces blood pressure in salt-induced hypertension. Cardiovasc Res 89(3):574–585.

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    McNally B, Griffin JL, Roberts LD (2016) Dietary inorganic nitrate: from villain to hero in metabolic disease? Mol Nutr Food Res 60(1):67–78.

    CAS  Article  PubMed  Google Scholar 

  9. 9.

    Butler AR, Feelisch M (2008) Therapeutic uses of inorganic nitrite and nitrate: from the past to the future. Circulation 117(16):2151–2159.

    CAS  Article  PubMed  Google Scholar 

  10. 10.

    Nossaman VE, Nossaman BD, Kadowitz PJ (2010) Nitrates and nitrites in the treatment of ischemic cardiac disease. Cardiol Rev 18(4):190–197.

    Article  PubMed  PubMed Central  Google Scholar 

  11. 11.

    King AM, Glass KA, Milkowski AL, Sindelar JJ (2015) Impact of clean-label antimicrobials and nitrite derived from natural sources on the outgrowth of clostridium perfringens during cooling of deli-style Turkey breast. J Food Prot 78(5):946–953.

    CAS  Article  PubMed  Google Scholar 

  12. 12.

    Al-Rasheed NM, Fadda LM, Attia HA, Ali HM, Al-Rasheed NM (2017) Quercetin inhibits sodium nitrite-induced inflammation and apoptosis in different rats organs by suppressing Bax, HIF1-α, TGF-β, Smad-2, and AKT pathways. J Biochem Mol Toxicol 31(5).

  13. 13.

    Chui JS, Poon WT, Chan KC, Chan AY, Buckley TA (2005) Nitrite-induced methaemoglobinaemia - aetiology, diagnosis and treatment. Anaesthesia 60(5):496–500.

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Ansari FA, Ali SN, Arif H, Khan AA, Mahmood R (2017) Acute oral dose of sodium nitrite induces redox imbalance, DNA damage, metabolic and histological changes in rat intestine. PLoS One 12(4):e0175196.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  15. 15.

    Ansari FA, Mahmood R (2016) Sodium nitrite enhances generation of reactive oxygen species that decrease antioxidant power and inhibit plasma membrane redox system of human erythrocytes. Cell Biol Int 40(8):887–894.

    CAS  Article  PubMed  Google Scholar 

  16. 16.

    Sherif IO, Al-Gayyar MM (2013) Antioxidant, anti-inflammatory and hepatoprotective effects of silymarin on hepatic dysfunction induced by sodium nitrite. Eur Cytokine Netw 24(3):114–121.

    CAS  Article  PubMed  Google Scholar 

  17. 17.

    Hazman Ö, Bozkurt MF, Fidan AF, Uysal FE, Çelik S (2018) The effect of boric acid and borax on oxidative stress, inflammation, ER stress and apoptosis in cisplatin toxication and nephrotoxicity developing as a result of toxication. Inflammation 41(3):1032–1048.

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Yamada KE, Eckhert CD (2019) Boric acid activation of eIF2α and Nrf2 is PERK dependent: a mechanism that explains how boron prevents DNA damage and enhances antioxidant status. Biol Trace Elem Res 188(1):2–10.

    CAS  Article  PubMed  Google Scholar 

  19. 19.

    Khaliq H, Juming Z, Ke-Mei P (2018) The physiological role of boron on health. Biol Trace Elem Res 186(1):31–51.

    CAS  Article  PubMed  Google Scholar 

  20. 20.

    Pawa S, Ali S (2006) Boron ameliorates fulminant hepatic failure by counteracting the changes associated with the oxidative stress. Chem Biol Interact 160(2):89–98.

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Uluisik I, Karakaya HC, Koc A (2018) The importance of boron in biological systems. J Trace Elem Med Biol 45:156–162.

    CAS  Article  PubMed  Google Scholar 

  22. 22.

    Tanaka M, Fujiwara T (2008) Physiological roles and transport mechanisms of boron: perspectives from plants. Pflugers Arch - Eur J Physiol 456(4):671–677.

    CAS  Article  Google Scholar 

  23. 23.

    Mahabir S, Spitz MR, Barrera SL, Dong YQ, Eastham C, Forman MR (2008) Dietary boron and hormone replacement therapy as risk factors for lung cancer in women. Am J Epidemiol 167(9):1070–1080.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  24. 24.

    Kot FS (2009) Boron sources, speciation and its potential impact on health. Rev Environ Sci Biotechnol 8(1):3–28.

  25. 25.

    Hacioglu C, Kar F, Kacar S, Sahinturk V, Kanbak G (2020) High concentrations of boric acid trigger concentration-dependent oxidative stress, apoptotic pathways and morphological alterations in DU-145 human prostate cancer cell line. Biol Trace Elem Res 193(2):400–409.

    CAS  Article  PubMed  Google Scholar 

  26. 26.

    Yildirim S, Celikezen FC, Oto G, Sengul E, Bulduk M, Tasdemir M, Ali Cinar D (2018) An investigation of protective effects of litium borate on blood and histopathological parameters in acute cadmium-induced rats. Biol Trace Elem Res 182(2):287–294.

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Ansari FA, Khan AA, Mahmood R (2018) Protective effect of carnosine and N-acetylcysteine against sodium nitrite-induced oxidative stress and DNA damage in rat intestine. Environ Sci Pollut Res Int 25(20):19380–19392.

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Ansari FA, Ali SN, Khan AA, Mahmood R (2018) Acute oral dose of sodium nitrite causes redox imbalance and DNA damage in rat kidney. J Cell Biochem 119(4):3744–3754.

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Ansari FA, Khan AA, Mahmood R (2018) Ameliorative effect of carnosine and N-acetylcysteine against sodium nitrite induced nephrotoxicity in rats. J Cell Biochem 120:7032–7044.

    CAS  Article  Google Scholar 

  30. 30.

    Soliman M, Aldhahrani A, Alkhedaide A, Nassan MA, Althobaiti F, Mohamed WA (2020) The ameliorative impacts of Moringa oleifera leaf extract against oxidative stress and methotrexate-induced hepato-renal dysfunction. Biomed Pharmacother 128:110259.

    CAS  Article  PubMed  Google Scholar 

  31. 31.

    Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95(2):351–358.

    CAS  Article  Google Scholar 

  32. 32.

    Beutler E, Duron O, Kelly BM (1963) Improved method for the determination of blood glutathione. J Lab Clin Med 61:882–888

    CAS  PubMed  Google Scholar 

  33. 33.

    Paglia DE, Valentine WN (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 70(1):158–169

    CAS  PubMed  Google Scholar 

  34. 34.

    Hadwan MH (2018) Simple spectrophotometric assay for measuring catalase activity in biological tissues. BMC Biochem 19(1):7.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  35. 35.

    Elsherbiny NM, Maysarah NM, El-Sherbiny M, Al-Gayyar MM (2017) Renal protective effects of thymoquinone against sodium nitrite-induced chronic toxicity in rats: impact on inflammation and apoptosis. Life Sci 180:1–8.

    CAS  Article  PubMed  Google Scholar 

  36. 36.

    Sifuentes-Franco S, Padilla-Tejeda DE, Carrillo-Ibarra S, Miranda-Díaz AG (2018) Oxidative stress, apoptosis, and mitochondrial function in diabetic nephropathy. Int J Endocrinol 2018:1875870–1875813.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  37. 37.

    Gyurászová M, Gurecká R, Bábíčková J, Tóthová Ľ (2020) Oxidative stress in the pathophysiology of kidney disease: implications for noninvasive monitoring and identification of biomarkers. Oxidative Med Cell Longev 2020:5478708–5478711.

    CAS  Article  Google Scholar 

  38. 38.

    Adewale OO, Samuel ES, Manubolu M, Pathakoti K (2019) Curcumin protects sodium nitrite-induced hepatotoxicity in Wistar rats. Toxicol Rep 6:1006–1011.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  39. 39.

    Kobylewski SE, Henderson KA, Yamada KE, Eckhert CD (2017) Activation of the EIF2α/ATF4 and ATF6 pathways in DU-145 cells by boric acid at the concentration reported in men at the US mean boron intake. Biol Trace Elem Res 176(2):278–293.

    CAS  Article  PubMed  Google Scholar 

  40. 40.

    Satta S, Mahmoud AM, Wilkinson FL, Yvonne Alexander M, White SJ (2017) The role of Nrf2 in cardiovascular function and disease. Oxidative Med Cell Longev 2017:9237263–9237218.

    CAS  Article  Google Scholar 

  41. 41.

    Mahmoud AM, Alexander MY, Tutar Y, Wilkinson FL, Venditti A (2017) Oxidative stress in metabolic disorders and drug-induced injury: the potential role of Nrf2 and PPARs activators. Oxidative Med Cell Longev 2017:2508909–2508904.

    Article  Google Scholar 

  42. 42.

    Pan H, Wang H, Wang X, Zhu L, Mao L (2012) The absence of Nrf2 enhances NF-κB-dependent inflammation following scratch injury in mouse primary cultured astrocytes. Mediat Inflamm 2012:217580–217589.

    CAS  Article  Google Scholar 

  43. 43.

    Baek JH, Zhang X, Williams MC, Hicks W, Buehler PW, D'Agnillo F (2015) Sodium nitrite potentiates renal oxidative stress and injury in hemoglobin exposed guinea pigs. Toxicology 333:89–99.

    CAS  Article  PubMed  Google Scholar 

  44. 44.

    Wardyn JD, Ponsford AH, Sanderson CM (2015) Dissecting molecular cross-talk between Nrf2 and NF-κB response pathways. Biochem Soc Trans 43(4):621–626.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  45. 45.

    Zhao K, Wen LB (2018) DMF attenuates cisplatin-induced kidney injury via activating Nrf2 signaling pathway and inhibiting NF-kB signaling pathway. Eur Rev Med Pharmacol Sci 22(24):8924–8931.

    CAS  Article  PubMed  Google Scholar 

  46. 46.

    Fadda LM, Attia HA, Al-Rasheed NM, Ali HM, Aldossari M (2017) Attenuation of DNA damage and mRNA gene expression in hypoxic rats using natural antioxidants. J Biochem Mol Toxicol 31(12).

  47. 47.

    Hu HH, Chen DQ, Wang YN, Feng YL, Cao G, Vaziri ND, Zhao YY (2018) New insights into TGF-β/Smad signaling in tissue fibrosis. Chem Biol Interact 292:76–83.

    CAS  Article  PubMed  Google Scholar 

  48. 48.

    Palanisamy N, Kannappan S, Anuradha CV (2011) Genistein modulates NF-κB-associated renal inflammation, fibrosis and podocyte abnormalities in fructose-fed rats. Eur J Pharmacol 667(1-3):355–364.

    CAS  Article  PubMed  Google Scholar 

  49. 49.

    Sutariya B, Saraf M (2017) Betanin, isolated from fruits of Opuntia elatior Mill attenuates renal fibrosis in diabetic rats through regulating oxidative stress and TGF-β pathway. J Ethnopharmacol 198:432–443.

    CAS  Article  PubMed  Google Scholar 

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We greatly appreciate and thank Taif University for the financial support for Taif University Researchers Supporting Project (TURSP-2020/09), Taif University, Taif, Saudi Arabia.

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Correspondence to Mohamed Mohamed Soliman.

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All experimental procedures were carried out under National Institutes of Health Guidelines for the care and use of laboratory animals, and all procedures designed to minimize the suffering of animals were followed.

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Soliman, M.M., Aldhahrani, A., Elshazly, S.A. et al. Borate Ameliorates Sodium Nitrite-Induced Oxidative Stress Through Regulation of Oxidant/Antioxidant Status: Involvement of the Nrf2/HO-1 and NF-κB Pathways. Biol Trace Elem Res (2021).

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  • Sodium nitrite
  • Oxidative stress
  • Molecular changes
  • Borat impacts