Effect of Exogenous Zinc on MsrB1 Expression and Protein Oxidation in Human Lens Epithelial Cells

  • Yi JiaEmail author
  • Jie Dai
  • Liangliang Zhang
  • Huan Xia


Aging has been related to zinc deficiency, resulting in protein oxidation and age-related decline of methionine sulfoxide reductase (Msr) activity. This study was designed to investigate the levels of methionine sulfoxide reductase B1 (MsrB1) mRNA and oxidized proteins in human lens epithelial (hLE) cells after treatment with exogenous zinc. The role of exogenous zinc in regulation of MsrB1 gene expression and protein oxidation in hLE cells was studied by MTT assay, oxidized protein measurement kit, and real-time PCR. The results showed that hLE cell viability was significantly decreased by MsrB1 gene knockdown or peroxynitrite (ONOO) treatment, while it was significantly increased after treatment with exogenous zinc (P < 0.05). Protein carbonyl content in hLE cell by MsrB1 gene knockdown or ONOO treatment was significantly decreased after treatment with ZnSO4 (P < 0.01). And exogenous zinc could increase the level of MsrB1 in hLE cell under normal (P < 0.001) and oxidative stress (P < 0.01) conditions. In conclusion, exogenous zinc could protect hLE cells against MsrB1 gene knockdown or ONOO-induced cell death by upregulation of MsrB1 involved in the elimination of reactive oxygen species (ROS) and oxidized proteins.


Methionine sulfoxide reductase B1 Protein oxidation Exogenous zinc 


Funding Information

This research was funded by the National Natural Science Foundation of China (No. 21561006) and the Science and Technology Foundation of Guizhou Province (No. LH-[2016]7372).


  1. 1.
    Kumar SD, Vijaya M, Samy RP, Dheen ST, Ren M, Watt F, Kang YJ, Bay BH, Tay SSW (2012) Zinc supplementation prevents cardiomyocyte apoptosis and congenital heart defects in embryos of diabetic mice. Free Radic Biol Med 53:1595–1606CrossRefPubMedGoogle Scholar
  2. 2.
    Zhang X, Wang J, Fan Y, Yang L, Wang L, Ma J (2012) Zinc supplementation attenuates high glucose-induced epithelial-to-mesenchymal transition of peritoneal mesothelial cells. Biol Trace Elem Res 150:229–235CrossRefPubMedGoogle Scholar
  3. 3.
    Prasad AS, Beck FWJ, Bao B, Fitzgerald JT, Snell DC, Steinberg JD, Cardozo LJ (2007) Zinc supplementation decreases incidence of infections in the elderly: effect of zinc on generation of cytokines and oxidative stress. Am J Clin Nutr 85:837–844CrossRefPubMedGoogle Scholar
  4. 4.
    Du Y, Guo D, Wu Q, Liu D, Bi H (2014) Zinc chloride inhibits human lens epithelial cell migration and proliferation involved in TGF-β1 and TNF-α signaling pathways in HLE B-3 cells. Biol Trace Elem Res 159(1–9):425–433CrossRefPubMedGoogle Scholar
  5. 5.
    Grahn BH, Paterson PG, Gottschall-Pass KT, Zhang Z (2001) Zinc and the eye. J Am Coll Nutr 20(2):106–118CrossRefPubMedGoogle Scholar
  6. 6.
    Shukla N, Moitra JK, Trivedi RC (1996) Determination of lead, zinc, potassium, calcium, copper and sodium in human cataract lenses. Sci Total Environ 181:161–165CrossRefPubMedGoogle Scholar
  7. 7.
    Barman S, Srinivasan K (2018) Zinc supplementation ameliorates diabetic cataract through modulation of crystallin proteins and polyol pathway in experimental rats. Biol Trace Elem Res.
  8. 8.
    Nourmohammadi I, Modarress M, Pakdel F (2006) Assessment of aqueous humor zinc status in human age-related cataract. Ann Nutr Metab 50:51–53CrossRefPubMedGoogle Scholar
  9. 9.
    Gunduz G, Gunduz F, Yucel I, Senturk UK (2003) Levels of zinc and magnesium in senile and diabetic senile cataractous lenses. Biol Trace Elem Res 95(2):107–112CrossRefPubMedGoogle Scholar
  10. 10.
    Lancellotti S, Filippis VD, Pozzi N, Peyvandi F, Palla R, Rocca B, Rutella S, Pitocco D, Mannucci PM, Cristofaro RD (2010) Formation of methionine sulfoxide by peroxynitrite at position 1606 of von Willebrand factor inhibits its cleavage by ADAMTS-13: a new prothrombotic mechanism in diseases associated with oxidative stress. Free Radic Biol Med 48:446–456CrossRefPubMedGoogle Scholar
  11. 11.
    Dyke KV, Ghareeb E, Dyke MV, Sosa A, Hoeldtke RD, Van Thiel DH (2008) Luminescence experiments involved in the mechanism of streptozotocin diabetes and cataract formation. Luminescence 23:386–391CrossRefPubMedGoogle Scholar
  12. 12.
    Garner MH, Spector A (1980) Selective oxidation of cysteine and methionine in normal and senile cataractous lenses. Proc Natl Acad Sci U S A 77:1274–1277CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Kantorow M, Hawse JR, Cowell TL, Benhamed S, Pizarro GO, Reddy VN, Hejtmancik JF (2004) Methionine sulfoxide reductase A is important for lens cell viability and resistance to oxidative stress. Proc Natl Acad Sci U S A 101:9654–9659CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Michael R, Bron AJ (2011) The ageing lens and cataract: a model of normal and pathological ageing. Philos Trans R Soc B 366:1278–1292CrossRefGoogle Scholar
  15. 15.
    Kim HY, Gladyshev VN (2004) Methionine sulfoxide reduction in mammals: characterization of methionine-R-sulfoxide reductases. Mol Biol Cell 15:1055–1064CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Brennan LA, Kantorow M (2009) Mitochondrial function and redox control in the aging eye: role of MsrA and other repair systems in cataract and macular degenerations. Exp Eye Res 88:195–203CrossRefPubMedGoogle Scholar
  17. 17.
    Marchetti MA, Pizarro GO, Sagher D, DeAmicis C, Brot N, Hejtmancik JF, Weissbach H, Kantorow M (2005) Methionine sulfoxide reductases B1, B2, and B3 are present in the human lens and confer oxidative stress resistance to lens cells. Invest Ophthalmol Vis Sci 46:2107–2112CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Hawse JR, Hejtmancik JF, Horwitz J, Kantorow M (2004) Identification and functional clustering of global gene expression differences between age-related cataract and clear human lenses and aged human lenses. Exp Eye Res 79:935–940CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Kumar RA, Koc A, Cerny RL, Gladyshev VN (2002) Reaction mechanism, evolutionary analysis, and role of zinc in drosophila methionine-R-sulfoxide reductase. J Biol Chem 277:37527–37535CrossRefPubMedGoogle Scholar
  20. 20.
    Picot CR, Perichon M, Lundberg KC, Friguet B, Szweda LI, Petropoulos I (2006) Alterations in mitochondrial and cytosolic methionine sulfoxide reductase activity during cardiac ischemia and reperfusion. Exp Gerontol 41:663–667CrossRefPubMedGoogle Scholar
  21. 21.
    Jia Y, Li Y, Du S, Huang K (2012) Involvement of MsrB1 in the regulation of redox balance and inhibition of peroxynitrite-induced apoptosis in human lens epithelial cells. Exp Eye Res 100:7–16CrossRefPubMedGoogle Scholar
  22. 22.
    Li Y, Zhang W, Li P, Huang K (2011) Effect of streptozocin-induced diabetes mellitus on expression of methionine sulfoxide reductases and accumulation of their substrates in mouse lenses. Exp Eye Res 92(5):401–407CrossRefPubMedGoogle Scholar
  23. 23.
    Li Y, Jia Y, Zhou J, Huang K (2013) Effect of methionine sulfoxide reductase B1 silencing on high-glucose-induced apoptosis of human lens epithelial cells. Life Sci 92:193–201CrossRefPubMedGoogle Scholar
  24. 24.
    Picot CR, Perichon M, Cintrat JC, Friguet B, Petropoulos I (2004) The peptide methionine sulfoxide reductases, MsrA and MsrB (hCBS-1), are downregulated during replicative senescence of human WI-38 fibroblasts. FEBS Lett 558:74–78CrossRefPubMedGoogle Scholar
  25. 25.
    Petropoulos I, Mary J, Perichon M, Friguet B (2001) Rat peptide methionine sulphoxide reductase: cloning of the cDNA, and down-regulation of gene expression and enzyme activity during aging. Biochem J 355:819–825CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Cabreiro F, Perichon M, Jatje J, Malavolta M, Mocchegiani E, Friguet B, Petropoulos I (2007) Zinc supplementation in the elderly subjects: effect on oxidized protein degradation and repair systems in peripheral blood lymphocytes. Exp Gerontol 43(5):483–487CrossRefPubMedGoogle Scholar
  27. 27.
    Klotz LO, Schieke SM, Sies H, Holbrook NJ (2000) Peroxynitrite activates the phosphoinositide 3-kinase/Akt pathway in human skin primary fibroblasts. Biochem J 352:219–225CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Zhou J, Li HD, Zeng JH, Huang KX (2009) Effects of peroxynitite-induced protein tyrosine nitration on insulin-stimulated tyrosine phosphorylation in HepG2 cells. Mol Cell Biochem 331:49–57CrossRefPubMedGoogle Scholar
  29. 29.
    Luca AD, Sacchetta P, Nieddu M, Ilio CD, Favaloro B (2007) Important roles of multiple Sp1 binding sites and epigenetic modifications in the regulation of the methionine sulfoxide reductase B1 (MsrB1) promoter. BMC Mol Biol 8:39–50CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Du S, Zhou J, Jia Y, Huang K (2010) SelK is a novel ER stress-regulated protein and protects HepG2 cells from ER stress agent-induced apoptosis. Arch Biochem Biophys 502:137–143CrossRefPubMedGoogle Scholar
  31. 31.
    Cao GH, Cutler RG (1995) Protein oxidation and aging. I. Difficulties in measuring reactive protein carbonyls in tissues using 2, 4-dinitrophenylhydrazine. Arch Biochem Biophys 320(1):106–114CrossRefPubMedGoogle Scholar
  32. 32.
    Chiou GC (2001) Effects of nitric oxide on eye diseases and their treatment. J Ocul Pharmacol Ther 17:189–198CrossRefPubMedGoogle Scholar
  33. 33.
    Hao LN, Ling YQ, Luo XM, Mao YX, Mao QY, He SZ, Ling YL (2006) Puerarin decreases lens epithelium cell apoptosis induced partly by peroxynitrite in diabetic rats. Acta Physiol Sinica 58(6):584–592PubMedGoogle Scholar
  34. 34.
    Li WC, Kuszak JR, Dunn K, Wang RR, Ma W, Wang GM, Spector A, Leib M, Cotlia AM, Weiss M (1995) Lens epithelial cell apoptosis appears to be a common cellular basis for non-congenital cataract development in humans and animals. J Cell Biol 130:169–181CrossRefPubMedGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Chemical Biology, School of Biology and EngineeringGuizhou Medical UniversityGuiyangPeople’s Republic of China

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