Abstract
Oxidative stress, as mediated by ROS (reactive oxygen species), is a significant factor in initiating the cells damaged by affecting cellular macromolecules and impairing their biological functions; SelX, a selenoprotein also known as MsrB1 belonging to the methionine sulfoxide reductase (Msr) family, is the redox repairing enzyme and involved in redox-related functions. In order to more precisely analyze the relationship between oxidative stress, cell oxidative damage, and SelX, we stably overexpressed porcine Selx full-length cDNA in human normal hepatocyte (LO2) cells. Cell viability, cell apoptosis rate, intracellular ROS, and the expression levels of mRNA or protein of apoptosis-related genes under H2O2-induced oxidative stress were detected. We found that overexpression of SelX can prevent the oxidative damage caused by H2O2 and propose that the main mechanism underlying the protective effects of SelX is the inhibition of LO2 cell apoptosis. The results revealed that overexpressed SelX reduced the H2O2-induced intracellular ROS generation, inhibited the H2O2-induced upregulation of Bax and downregulation of Bcl-2, and increased the mRNA and protein ratio of Bcl-2/Bax. Furthermore, it inhibited H2O2-induced p38 MAPK phosphorylation. Taken together, our findings suggested that SelX played important roles in protecting LO2 cells against oxidative damage and that its protective effect is partly via the p38 pathway by acting as a ROS scavenger.
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Abbreviations
- Msr:
-
Methionine sulfoxide reductase
- SelX:
-
Selenoprotein X
- ROS:
-
Reactive oxygen species
- LO2:
-
Human normal hepatocyte;
- p38 MAPK:
-
p38 mitogen-activated protein kinase
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- DMSO:
-
Dimethyl sulfoxide
- ECL:
-
Electrochemiluminescence
- PVDF:
-
Polyvinylidene fluoride
- SDS:
-
Sodium dodecyl sulfate
- SE:
-
Standard error
References
Liochev SI (2013) Reactive oxygen species and the free radical theory of aging. Free Radic Biol Med 60:1–4
Berlett BS, Stadtman ER (1997) Protein oxidation in aging, disease, and oxidative stress. J Biol Chem 272:20313–20316
Zhang JX, Wang XL, Vikash V, Ye Q, Wu DD, Liu YL (2016) ROS and ROS-mediated cellular signaling. Oxid Med Cell Longe 4350965
Ozsurekci Y, Aykac K (2016) Oxidative stress related diseases in newborns. Oxid Med Cell Longe 2768365
Leonard SS, Harris GK, Shi X (2004) Metal-induced oxidative stress and signal transduction. Free Radic Biol Med 37:1921–1942
Ravingerová T, Barancík M, Strnisková M (2003) Mitogen activated protein kinase: a new therapeutic target in cardiac pathology. Mol Cell Biochem 247:127–138
Abdel-Misih SR, Bloomston M (2010) Liver anatomy. Surg Clin N Am 90:643–653
Burk RF, Hill KE (2005) Selenoprotein P: an extracellular protein with unique physical characteristics and a role in selenium homeostasis. Annu Rev Nutr 25:215–235
Maton A, Hopkins J, Charles WM, Susan M, Maryanna QW, David L, Jill DW (1993) Human biology and health. Prentice Hall, Englewood Cliffs
Simon HU, Haj-Yehia A, Levi-Schaffer F (2000) Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis 5:415–418
Patel T, Gores GJ (1995) Apoptosis and hepatobiliary disease. Hepatology 21:1725–1741
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 41:663–667
Han YH, Zhang ZW, Su J, Zhang B, Li S, Xu SW (2012) Effects of chicken selenoprotein W on H2O2-induced apoptosis in CHO-K1 cells. Biol Trace Elem Res 147:395–402
Chen W, Liu Y, Xue G, Zhang L, Zhang L, Shao S (2016) Diazoxide protects L6 skeletal myoblasts from H2O2-induced apoptosis via the phosphatidylinositol-3 kinase/Akt pathway. Inflamm Res 65:53–60
Tang JY, Cao L, Li Q, Wang LQ, Jia G, Liu GM, Chen XL, Cai JY, Shang HY, Zhao H (2016) Selenoprotein X gene knockdown aggravated H2O2-induced apoptosis in liver LO2 cells. Biol Trace Elem Res 173:71–78
Morris D, Khurasany M, Nguyen T, Kim J, Guilford F, Mehta R, Gray D, Saviola B, Venketaraman V (2013) Glutathione and infection, BBA-Gen. Subjects 1830:3329–3349
Papp LV, Holmgren A, Khanna KK (2010) Selenium and selenoproteins in health and disease. Antioxid Redox Sign 12:793–795
Lei XG, Cheng WH (2005) New roles for an old selenoenzyme: evidence from glutathione peroxidase-1 null and overexpressing mice. J Nutr 135:2295–2298
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 Ophth Vis Sci 46:2107–2112
Lee BC, Dikiy A, Kim HY, Gladyshev VN (2009) Functions and evolution of selenoprotein methionine sulfoxide reductases. Biochim Biophys Acta 1790:1471–1477
Kim HY, Gladyshev VN (2004) Characterization of mouse endoplasmic reticulum methionine-R-sulfoxide reductase. Biochem Bioph Res Co 320:1277–1283
Moghadaszadeh B, Beggs AH (2006) Selenoproteins and their impact on human health through diverse physiological pathways. Physiology 21:307–315
Levine RL, Mosoni L, Berlett BS, Stadtman ER (1996) Methionine residues as endogenous antioxidants in proteins. Proc Natl Acad Sci U S A 93:15036–15040
Jia Y, Zhou J, Liu HM, Huang KX (2014) Effect of methionine sulfoxide reductase B1 (SelR) gene silencing on peroxynitrite-induced F-actin disruption in human lens epithelial cells. Biochem Bioph Res Co 443:876–881
Jia Y, Li Y, Du S, Huang KX (2012) Involvement of MsrB1 in the regulation of redox balance and inhibition of peroxynitrite-induced apoptosis in human lens epithelial cells. Exp Eye Res 100:7–16
Zhou YJ, Zhang SP, Liu CW, Cai YQ (2009) The protection of selenium on ROS mediated-apoptosis by mitochondria dysfunction in cadmium-induced LLC-PK1 cells. Toxicol in Vitro 23:288–294
Liu Y, Zhao H, Zhang QS, Tang JY, Li K, Xia XJ, Wang KN, Li K, Lei XG (2012) Prolonged dietary selenium deficiency or excess does not globally affect selenoprotein gene expression and/or protein production in various tissues of pigs. J Nutr 142:1410–1416
Huang XF, Tang JY, Xu JY, Jia G, Liu GM, Chen XL, Cai JY, Shang HY, Zhao H (2016) Supranutritional dietary selenium induced hyperinsulinemia and dyslipidemia via affected expression of selenoprotein genes and insulin signal-related genes in broiler. RSC Adv 6:84990–84998
Zhao H, Tang JY, Cao L, Zhou JC, Jia G, Liu GM, Chen XL, Wang KN (2015) Gene cloning, site-directed mutagenesis, prokaryotic expression and polyclonal antibody preparation of porcine selenoprotein X. Chinese Journal of Animal Nutrition 27:1485–1491
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63
Oien DB, Moskovitz J (2009) Selenium and the methionine sulfoxide reductase system. Molecules 14:2337–2344
Picot CR, Petropoulos I, Perichon M, Moreau M, Nizard C, Friguet B (2005) Overexpression of MsrA protects WI-38 SV40 human fibroblasts against H2O2-mediated oxidative stress. Free Radic Biol Med 39:1332–1341
Li Y, Jia Y, Zhou J, Huang KX (2013) Effect of methionine sulfoxide reductase B1 silencing on high-glucose-induced apoptosis of human lens epithelial cells. Life Sci 92:193–201
Fomenko DE, Novoselov SV, Natarajan SK, Lee BC, Koc A, Carlson BA, Lee TH, Kim HY, Hatfield DL, Gladyshev VN (2009) MsrB1 (methionine-R-sulfoxide reductase 1) knock-out mice: roles of MsrB1 in redox regulation and identification of a novel selenoprotein form. J Biol Chem 284:5986–5993
Manuele MG, Arcos MLB, Davicino R, Ferraro G, Cremaschi G, Anesini C (2010) Limonene exerts antiproliferative effects and increases nitric oxide levels on a lymphoma cell line by dual mechanism of the ERK pathway: relationship with oxidative stress. Cancer Investig 2:135–145
Varma SD, Kovtun S, Hegde KR (2011) Role of ultraviolet irradiation and oxidative stress in cataract formation-medical prevention by nutritional antioxidants and metabolic agonists. Eye Contact Lens 37:233–245
Franklin JL (2011) Redox regulation of the intrinsic pathway in neuronal apoptosis. Antioxid Redox Sign 14:1437–1448
Zhang H, Gomez AM, Wang X, Yan Y, Zheng M, Cheng H (2013) ROS regulation of microdomain Ca2+ signalling at the dyads. Cardiovasc Res 98:248–258
Sena LA, Chandel NS (2012) Physiological roles of mitochondrial reactive oxygen species. Mol Cell 48:158–166
Vander Heiden MG, Chandel NS, Williamson EK, Schumacker PT, Thompson CB (1997) Bcl-xL regulates the membrane potential and volume homeostasis of mitochondria. Cell 91:627–637
Petit A, Mwale F, Zukor DJ, Catelas I, Antoniou J, Huk OL (2004) Effect of cobalt and chromium ions on bcl-2, bax, caspase-3, and caspase-8 expression in human U937 macrophages. Biomaterials 25:2013–2018
Bai J, Zheng Y, Wang G, Liu P (2016) Protective effect of D-Limonene against oxidative stress-induced cell damage in human lens epithelial cells via the p38 pathway. Oxid Med Cell Longe: 5962832
Thornberry NA, Lazebnik Y (1998) Caspases: enemies within. Science 281:1312–1316
Kim DS, Kim JH, Lee GH, Kim HT, Lim JM, Chae SW, Chae HJ, Kim HR (2010) p38 mitogen-activated protein kinase is involved in endoplasmic reticulum stress induced cell death and autophagy in human gingival fibroblasts. Biol Pharm Bull 33:545–549
Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME (1995) Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 270:1326–1331
Liu BR, Yuan B, Zhang L, Mu WM, Wang CM (2015) ROS/p38/p53/Puma signaling pathway is involved in emodin-induced apoptosis of human colorectal cancer cells. Int J Clin Exp Med 8:15413–15422
Cardaci S, Filomeni G, Rotilio G, Ciriolo MR (2010) p38 (MAPK)/p53 signalling axis mediates neuronal apoptosis in response to tetrahydrobiopterin-induced oxidative stress and glucose uptake inhibition: implication for neurodegeneration. Biochem J 430:439–451
Bai J, Zheng Y, Dong L, Cai XH, Wang G, Liu P (2015) Inhibition of p38 mitogen-activated protein kinase phosphorylation decreases H2O2-induced apoptosis in human lens epithelial cells. Graef Arch Clin Exp 253:1933–1940
Kim JY, Lee JS, Han YS, Lee JH, Bae I, Yoon YM, Kwon SM, Lee SH (2015) Pretreatment with lycopene attenuates oxidative stress-induced apoptosis in human mesenchymal stem cells. Biomol Ther 23:517–524
Acknowledgements
This work was supported partly by the National Natural Science Foundation of China (No 31272468 and 31072043) and by a Research Funding provided by Sichuan Longda animal husbandry science and technology co., Ltd. (No: 2015SCLD001).
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All authors read and approved the final manuscript. The authors have no conflicts of interest to disclose.
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Ai-Hua He is the co-first author.
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Tang, JY., He, AH., Jia, G. et al. Protective Effect of Selenoprotein X Against Oxidative Stress-Induced Cell Apoptosis in Human Hepatocyte (LO2) Cells via the p38 Pathway. Biol Trace Elem Res 181, 44–53 (2018). https://doi.org/10.1007/s12011-017-1025-z
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DOI: https://doi.org/10.1007/s12011-017-1025-z