Molecular and Cellular Biochemistry

, Volume 360, Issue 1–2, pp 133–145 | Cite as

Repeated short-term stress synergizes the ROS signalling through up regulation of NFkB and iNOS expression induced due to combined exposure of trichloroethylene and UVB rays



Restraint stress is known to catalyse the pathogenesis of the variety of chronic inflammatory disorders. The present study was designed to evaluate the effect of repeated short-term stress (RRS) on cellular transduction apart from oxidative burden and early tumour promotional biomarkers induced due to combined exposure of trichloroethylene (TCE) and Ultra-violet radiation (UVB). RRS leads to the increase in the expression of the stress responsive cellular transduction elements NFkB-p65 and activity of iNOS in the epidermal tissues of mice after toxicant exposure. RRS augments the steep depletion of the cellular antioxidant machinery which was evidenced by the marked depletion in GSH (Glutathione and GSH dependant enzymes), superoxide dismutase and catalase activity that were observed at significance level of P < 0.001 with increase in lipid peroxidation, H2O2 and xanthine oxidase activity (P < 0.001) in the stressed animals and down regulation of DT-diaphorase activity (P < 0.001). Since, the induction of NFkB-p65 and inducible nitric oxide synthase expression mediated can lead to the hyperproliferation, we estimated a significant increment (P < 0.001) in the synthesis of polyamines in mice skin evidenced here by the ornithine decarboxylase which is the early marker of tumour promotion and further evaluated PCNA expression. All these findings cues towards the synergising ability of repeated short-term stress in the toxic response of TCE and UVB radiation.


Repeated restraint stress NFkB-p65 iNOS ODC PCNA 



Repeated restraint stress




Ultra-violet radiation


Ornithine decarboxylase


Nuclear factor kappa B


Inducible nitric oxide synthase




Glutathione reductase


Glutathione peroxidase


Superoxide dismutase




Lipid peroxidation


Hydrogen peroxide


Xanthine oxidase


Proliferating cell nuclear antigen




Nicotinamide adenine dinucleotide phosphate





Author Sarwat Sultana is thankful to University Grants Commission, Govt. of India, New Delhi, for providing Meritorious Research Fellowship to her student Farrah Ali.

Conflict of interest

Authors state no conflict of interest.


  1. 1.
    Bauer ME (2008) Chronic stress and immunosenescence: a review. Neuroimmunomodulation 15(4–6):241–250PubMedCrossRefGoogle Scholar
  2. 2.
    Elenkov IJ, Chrousos GP (2006) Stress system–organization, physiology and immunoregulation. Neuroimmunomodulation 13(5–6):257–267PubMedCrossRefGoogle Scholar
  3. 3.
    Dragoş D, Tănăsescu MD (2010) The effect of stress on the defense systems. J Med Life 3(1):10–18PubMedGoogle Scholar
  4. 4.
    Forsythe P, Ebeling C, Gordon JR, Befus AD, Vliagoftis H (2004) Opposing effects of short- and long-term stress on airway inflammation. Am J Respir Crit Care Med 169(2):220–226PubMedCrossRefGoogle Scholar
  5. 5.
    Zhang Y, Zhang Y, Miao J, Hanley G, Stuart C, Sun X, Chen T, Yin D (2008) Chronic restraint stress promotes immune suppression through toll-like receptor 4-mediated phosphoinositide 3-kinase signaling. J Neuroimmunol 204(1–2):13–19PubMedCrossRefGoogle Scholar
  6. 6.
    Dhabhar FS, Saul AN, Daugherty C, Holmes TH, Bouley DM, Oberyszyn TM (2010) Short-term stress enhances cellular immunity and increases early resistance to squamous cell carcinoma. Brain Behav Immun 24(1):127–137PubMedCrossRefGoogle Scholar
  7. 7.
    Dhabhar FS (2000) Acute stress enhances while chronic stress suppresses skin immunity. The role of stress hormones and leukocyte trafficking. Ann NY Acad Sci 917:876–893PubMedCrossRefGoogle Scholar
  8. 8.
    Segerstrom SC, Miller GE (2004) Psychological stress, the human immune system: a meta-analytic study of 30 years of inquiry. Psychol Bull 130(4):601–630PubMedCrossRefGoogle Scholar
  9. 9.
    Muqbil I, Azmi AS, Banu N (2006) Prior exposure to restraint stress enhances 7,12-dimethylbenz(a)anthracene (DMBA) induced DNA damage in rats. FEBS Lett 580(16):3995–3999PubMedCrossRefGoogle Scholar
  10. 10.
    Muqbil I, Banu N (2006) Enhancement of pro-oxidant effect of 7,12-dimethylbenz(a)anthracene (DMBA) in rats by pre-exposure to restraint stress. Cancer Lett 240(2):213–220PubMedCrossRefGoogle Scholar
  11. 11.
    Gidron Y, Ronson A (2008) Psychosocial factors, biological mediators, and cancer prognosis: a new look at an old story. Curr Opin Oncol 20(4):386–392PubMedCrossRefGoogle Scholar
  12. 12.
    Stojanovich L (2010) Stress and autoimmunity. Autoimmun Rev 9(5):A271–A276PubMedCrossRefGoogle Scholar
  13. 13.
    Aberg KM, Radek KA, Choi EH, Kim DK, Demerjian M, Hupe M, Kerbleski J, Gallo RL, Ganz T, Mauro T, Feingold KR, Elias PM (2007) Psychological stress down regulates epidermal antimicrobial peptide expression and increases severity of cutaneous infections in mice. J Clin Invest 117(11):3339–3349PubMedCrossRefGoogle Scholar
  14. 14.
    Dhabhar FS, McEwen BS (1999) Enhancing versus suppressive effects of stress hormones on skin immune function. Proc Natl Acad Sci USA 96(3):1059–1064PubMedCrossRefGoogle Scholar
  15. 15.
    Garg A, Chren MM, Sands LP, Matsui MS, Marenus KD, Feingold KR, Elias PM (2001) Psychological stress perturbs epidermal permeability barrier homeostasis: implications for the pathogenesis of stress-associated skin disorders. Arch Dermatol 137(1):53–59PubMedGoogle Scholar
  16. 16.
    Goel G, Makkar HP, Francis G, Becker K (2007) Phorbol esters: structure, biological activity, and toxicity in animals. Int J Toxicol 26(4):279–288PubMedCrossRefGoogle Scholar
  17. 17.
    Fujiki H, Suganuma M (2009) Carcinogenic aspects of protein phosphatase 1 and 2A inhibitors. Prog Mol Subcell Biol 46:221–254PubMedCrossRefGoogle Scholar
  18. 18.
    Lu J, Rho O, Wilker E, Beltran L, Digiovanni J (2007) Activation of epidermal akt by diverse mouse skin tumor promoters. Mol Cancer Res 5(12):1342–1352PubMedCrossRefGoogle Scholar
  19. 19.
    Barthelman M, Chen W, Gensler HL, Huang C, Dong Z, Bowden GT (1998) Inhibitory effects of perillyl alcohol on UVB-induced murine skin cancer and AP-1 transactivation. Cancer Res 15;58(4):711–716Google Scholar
  20. 20.
    Nishigori C, Hattori Y, Toyokuni S (2004) Role of reactive oxygen species in skin carcinogenesis. Antioxid Redox Signal 6(3):561–570PubMedCrossRefGoogle Scholar
  21. 21.
    Briones TL (2007) Psychoneuroimmunology and related mechanisms in understanding health disparities in vulnerable populations. Annu Rev Nurs Res 25:219–256PubMedGoogle Scholar
  22. 22.
    Choi EH, Brown BE, Crumrine D, Chang S, Man MQ, Elias PM, Feingold KR (2005) Mechanisms by which psychologic stress alters cutaneous permeability barrier homeostasis and stratum corneum integrity. J Invest Dermatol 124(3):587–595PubMedCrossRefGoogle Scholar
  23. 23.
    Gerlini G, Romagnoli P, Pimpinelli N (2005) Skin cancer and immunosuppression. Crit Rev Oncol Hematol 56(1):127–136PubMedCrossRefGoogle Scholar
  24. 24.
    Shen T, Zhu QX, Yang S, Wu CH, Zhang HF, Zhou CF, Zhang XJ (2008) Trichloroethylene induced cutaneous irritation in BALB/c hairless mice: histopathological changes and oxidative damage, Toxicology; 248(2–3:113–120Google Scholar
  25. 25.
    Swarbrick O (1965) The correction of prolapsed rectum and scrotal hernia in young pigs using trichloroethylene anaesthesia. Vet Rec 77(34):981–984PubMedCrossRefGoogle Scholar
  26. 26.
    Bakke B, Stewart PA, Waters MA (2007) Uses of and exposure to trichloroethylene in U.S. industry: a systematic literature review. Occup Environ Hyg 4(5):375–390CrossRefGoogle Scholar
  27. 27.
    Anagnostopoulos G, Sakorafas GH, Grigoriadis K, Margantinis G, Kostopoulos P, Tsiakos S, Arvanitidis D (2004) Hepatitis caused by occupational chronic exposure to trichloroethylene. Acta Gastroenterol Belg 67(4):355–357PubMedGoogle Scholar
  28. 28.
    Zhu QX, Ma T, Shen T, Yu Y, Ye LP, Ding R, Yu JF (2007) Cytotoxicity of trichloroethylene in keratinocytes involving alterations of mitochondrial function and ultrastructure. Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi 25(5):263–266Google Scholar
  29. 29.
    Kamijima M, Wang H, Huang H, Li L, Shibata E, Lin B, Sakai K, Liu H, Tsuchiyama F, Chen J, Okamura A, Huang X, Hisanaga N, Huang Z, Ito Y, Takeuchi Y, Nakajima T (2008) Trichloroethylene causes generalized hypersensitivity skin disorders complicated by Hepatitis. J Occup Health 50(4):328–338PubMedCrossRefGoogle Scholar
  30. 30.
    Kumar P, Prasad AK, Maji BK, Mani U, Dutta KK (2001) Hepatoxic alterations induced by inhalation of trichlorethylene (TCE) in rats. Biomed Environ Sci 14(4):325–326PubMedGoogle Scholar
  31. 31.
    Khan S, Priyamvada S, Khan SA, Khan W, Farooq N, Khan F, Yusufi AN (2009) Effect of trichloroethylene (TCE) toxicity on the enzymes of carbohydrate metabolism, brush border membrane and oxidative stress in kidney and other rat tissues. Food Chem Toxicol 47(7):1562–1568PubMedCrossRefGoogle Scholar
  32. 32.
    Shen T, Zhu QX, Yang S, Ding R, Ma T, Ye LP, Wang LJ, Liang ZZ, Zhang XJ (2007) Trichloroethylene induces nitric oxide production and nitric oxide synthase mRNA expression in cultured normal human epidermal keratinocytes. Toxicology 8;239(3):186–194Google Scholar
  33. 33.
    Molho-Pessach V, Lotem M (2007) Ultraviolet radiation and cutaneous carcinogenesis. Curr Probl Dermatol 35:14–27PubMedCrossRefGoogle Scholar
  34. 34.
    Epstein JH (1988) Photocarcinogenesis promotion studies with benzoyl peroxide (BPO) and croton oil. J Invest Dermatol 91(2):114–116PubMedCrossRefGoogle Scholar
  35. 35.
    Sharma S, Khan N, Sultana S (2005) Balsamodendron mukul suppresses benzoyl peroxide and ultraviolet light induced tumor promotional events in Swiss mice. J Photochem Photobiol B 14;78(1):43–51Google Scholar
  36. 36.
    Picardo M, Zompetta C, De Luca C, Amantea A, Faggioni A, Nazzaro-Porro M, Passi S (1991) Squalene peroxides may contribute to ultraviolet light-induced immunological effects. Photodermatol Photoimmunol Photomed 8(3):105–110PubMedGoogle Scholar
  37. 37.
    Saul AN, Oberyszyn TM, Daugherty C, Kusewitt D, Jones S, Jewell S, Malarkey WB, Lehman A, Lemeshow S, Dhabhar FS (2005) Chronic stress and susceptibility to skin cancer. J Natl Cancer Inst 97(23):1760–1767PubMedCrossRefGoogle Scholar
  38. 38.
    Sander CS, Chang H, Hamm F, Elsner P, Thiele JJ (2004) Role of oxidative stress and the antioxidant network in cutaneous carcinogenesis. Int J Dermatol 43:326–335PubMedCrossRefGoogle Scholar
  39. 39.
    Bradley PP, Priebat DA, Christensen RD, Rothstein G (1982) Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. J Invest Dermatol 78:206–209PubMedCrossRefGoogle Scholar
  40. 40.
    Wright JR, Colby HD, Miles PR (1981) Cytosolic factors which affect microsomal lipid peroxidation in lung and liver. Arch Biochem Biophys 206(2):296–304PubMedCrossRefGoogle Scholar
  41. 41.
    Pick E, Keisari Y (1980) A simple colorimetric method for the measurement of hydrogen peroxide produced by cells in culture. J Immunol Methods 38:161–170PubMedCrossRefGoogle Scholar
  42. 42.
    Stripe F, Della-Corte E (1969) The regulation of rat liver xanthine oxidase. Conversion in vitro of enzyme activity from dehydrogenase (type D) to oxidase (type O). J Biol Chem 244:3855–3863Google Scholar
  43. 43.
    Jollow DJ, Mitchell JR, Zampaglione N, Gillette JR (1974) Bromobenzene-induced liver necrosis. Protective role of glutathione and evidence for 3,4-bromobenzene oxide as the hepatotoxic metabolite. Pharmacology 11(3):151–169PubMedCrossRefGoogle Scholar
  44. 44.
    Mohandas J, Marshall JJ, Duggin GG, Horvath JS, Tiller DL (1984) Low activities of glutathione-related enzymes as factors in the genesis of urinary bladder cancer. Cancer Res 44:5086–5091PubMedGoogle Scholar
  45. 45.
    Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferase. The first enzymatic step in mercapturic acid formation. J. Biol Chem 249(22):7130–7139Google Scholar
  46. 46.
    Claiborne (1985) Catalase activity. In: Greenwald RA (ed) Handbook of methods for oxygen radical research. CRC press, Boca Raton, FL, pp 283–284Google Scholar
  47. 47.
    Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474PubMedCrossRefGoogle Scholar
  48. 48.
    Benson A, Hunkeler M, Talalay P (1980) Increase of NADPH; quinone reductase activity by dietary antioxidant, possible role in protection against carcinogenesis and toxicity. Proct Natl Acad Sci USA 5116–5620Google Scholar
  49. 49.
    O’Brien TG, Saladik D, Diamond L (1982) Effects of tumor-promoting phorbol diesters on neoplastic progression of Syrian hamster embryo cells. Cancer Res 42(4):1233–1238PubMedGoogle Scholar
  50. 50.
    Pence BC, Reiners JJ Jr (1987) Murine epidermal xanthine oxidase hyperplasia induced by tumor promoters. Cancer Res 47:6388–6392PubMedGoogle Scholar
  51. 51.
    Bhutia SK, Mallick SK, Maiti TK (2010) Tumour escape mechanisms and their therapeutic implications in combination tumour therapy. Cell Biol Int 34(5):553–563PubMedCrossRefGoogle Scholar
  52. 52.
    Whiteside TL (2006) Immune suppression in cancer: effects on immune cells, mechanisms and future therapeutic intervention. Semin Cancer Biol 16(1):3–15PubMedCrossRefGoogle Scholar
  53. 53.
    Ball TM (2006) Cortisol circadian rhythms and stress responses in infants at risk of allergic disease. Neuroimmunomodulation 13(5–6):294–300PubMedCrossRefGoogle Scholar
  54. 54.
    Bickers DR, Athar M (2006) Oxidative stress in the pathogenesis of skin disease. J Invest Dermatol 126(12):2565–2575PubMedCrossRefGoogle Scholar
  55. 55.
    Arck PC, Slominski A, Theoharides TC, Peters EM, Paus R (2006) Neuroimmunology of stress: skin takes center stage. J Invest Dermatol 126(8):1697–1704PubMedCrossRefGoogle Scholar
  56. 56.
    Mizobe K, Kishihara K, Ezz-Din El-Naggar R, Madkour GA, Kubo C, Nomoto K (1997) Restraint stress-induced elevation of endogenous glucocorticoid suppresses migration of granulocytes and macrophages to an inflammatory locus. J Neuroimmunol 73(1–2):81–89PubMedCrossRefGoogle Scholar
  57. 57.
    Rausbaugh HJ, Dallman MF, Levine JD (1999) Repeated, but not acute, stress suppresses inflammatory plasma extravasation. Proc Natl Acad Sci USA 96(25):14629–14634CrossRefGoogle Scholar
  58. 58.
    Butterfield TA, Best TM, Merrick MA (2006) The dual roles of neutrophils and macrophages in inflammation: a critical balance between tissue damage and repair. J Athl Train 41(4):457–465PubMedGoogle Scholar
  59. 59.
    Song SH, Min HY, Han AR, Nam JW, Seo EK, Park SW, Lee SH, Lee SK (2009) Suppression of inducible nitric oxide synthase by (−)-isoeleutherin from the bulbs of Eleutherine americana through the regulation of NF-kappaB activity. Int Immunopharmacol 9(3):298–302PubMedCrossRefGoogle Scholar
  60. 60.
    Sun Y (1990) Free radicals, antioxidant enzymes, and carcinogenesis. Free Radic Biol Med 8:583–590PubMedCrossRefGoogle Scholar
  61. 61.
    Ernster L (1967) DT-diaphorase. Methods Enzymol (10):309–317Google Scholar
  62. 62.
    Edwards YH, Potter J, Hopkinson DA (1980) Human FAD-dependant NADPH diaphorase. Biochem J 187:429–436PubMedGoogle Scholar
  63. 63.
    Schipper RG, Verhofstad AA (2002) Distribution patterns of ornithine decarboxylase in cells and tissues: facts, problems, and postulates. J Histochem Cytochem 50(9):1143–1160PubMedCrossRefGoogle Scholar
  64. 64.
    Kushlinskii NE, Orinovskii MB, Gurevich LE, Kazantseva IA, ShZh Talaeva, Shirokii VP, Ermilova VD, Dvorova EK, Ozherelev AS (2004) Expression of biomolecular markers (Ki-67, PCNA, Bcl-2, BAX, BclX, VEGF) in breast tumors. Bull Exp Biol Med 137(2):182–185PubMedCrossRefGoogle Scholar
  65. 65.
    Zhao H, Lo YH, Ma L, Waltz SE, Gray JK, Hung MC, Wang SC (2011) Targeting tyrosine phosphorylation of PCNA inhibits prostate cancer grow. Mol Cancer Ther 10(1):29–36PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  1. 1.Section of Molecular Carcinogenesis and Chemoprevention Toxicology, Department of Medical Elementology and Toxicology, Faculty of ScienceJamia Hamdard (Hamdard University)New DelhiIndia

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