Abstract
Acrolein, a major component of cigarette smoke, an environmental pollutant and an endogenous lipid peroxidation product, has been implicated in the development of atherosclerosis. Although a link between vascular injury and acrolein has been indicated, the exact molecular mechanism of acrolein-induced toxicity to vasculature is unknown. In an effort to elucidate the molecular basis of acrolein-induced vascular toxicity, the possibility of the intracellular signaling system as one of the targets of acrolein-induced toxicity is investigated in the present study. Exposure of cultured rat vascular smooth muscle cells (VSMCs) to different doses of acrolein not only causes cytotoxicity but also alters cellular morphology in a concentration and time-dependent manner. VSMCs exhibit cytotoxicity to a narrow concentration range of 5–10 μg/ml and display no toxicity to 2 μg/ml acrolein even after 24 h of exposure. Furthermore, exposure to acrolein results in activation of members of the mitogen-activated protein kinase (MAPK) family and protein tyrosine kinases. The extracellular signal-regulated kinases 1 and 2 (ERK1/2), stress-activated protein kinases/c-jun NH2-terminal kinases (SAPK/JNK) and p38MAPK are effectively and transiently activated by acrolein in a concentration and time-dependent fashion. While all three MAPKs exhibit significant activation within 5 min of exposure to acrolein, maximum activation (ERK1/2 and p38MAPK) or close to maximum activation (SAPK/JNK) occurs on exposure to 5 μg/ml acrolein for 2 h. Acrolein-induced activation of MAPKs is further substantiated by the activation of transcription factors, c-jun and activator transcription factor-2 (ATF-2), by acrolein-activated SAPK/JNK and p38MAPK, respectively. Additionally several cellular proteins exhibit spectacular protein tyrosine phosphorylation, particularly in response to 2 and 5 μg/ml of acrolein. Interestingly, the acrolein-induced activation of MAPKs precedes acrolein-stimulated protein tyrosine phosphorylation, which occurs after 2 h of exposure to acrolein. However, the time course of maximum protein tyrosine phosphorylation profile corresponds to the peak activation profile of MAPKs. The activation of MAPKs and protein tyrosine phosphorylation by acrolein appears to be independent of acrolein-induced toxicity. VSMCs exposed to 2 μg/ml acrolein exhibit no toxicity but stimulates significant activation of MAPKs and protein tyrosine phosphorylation. Although acrolein-induced VSMC toxicity is not blocked by MAPK inhibitors, PD98059, an inhibitor of MAPK kinase and SB203580, an inhibitor of p38MAPK, either alone or in combination, each MAPK responds differently to the inhibitors. Most prominently, although SB203580, an inhibitor of both SAPK/JNK and p38MAPK, significantly inhibited acrolein-induced activation of p38MAPK, it also stimulated SAPK/JNK activation by acrolein alone and in combination with PD98059. These results provide the first evidence that the activation of both growth-regulated (ERK1/2) and stress-regulated (SAPK/JNK and p38MAPK) MAPKs as well as tyrosine kinases are involved in the mediation of acrolein-induced effects on VSMC, which may play a crucial role in vascular pathogenesis due to environmentally and endogenously produced acrolein.
Similar content being viewed by others
References
Kunsch C, Medford RM: Oxidative stress as a regulator of gene expression in the vasculature. Circ Res 85: 753-766, 1999
Jacobs DR, Adachi H, Mulder I, Kromhout D, Menotti A, Nissinen A, Blackburn H: Cigarette smoking and mortality risk: Twenty-five-year follow-up of the seven countries study. Arch Intern Med 159: 733-740, 1999
Kannel WB: Update on the role of cigarette smoking in coronary artery disease. Am Heart J 101: 318-328, 1981
United States Department of Health and Human Services, Public Health Service Office: The health consequences of smoking, cancer. A report of the Surgeon General: Vapor phase components, cancer. Smoking and Health, 1982, pp 192-197
Syracuse Research Corporation: Toxicological Profile for Acrolein. Department of Health and Human Services, Agency for Toxic Substances and Disease Registry, Washington DC, 1990
World Health Organization: Environmental health criteria. In: Acrolein (Pub 127). World Health Organization, Geneva, 1991
Lash LH, Woods EB: Cytotoxicity of alkylating agents in isolated rat kidney proximal tubular and distal tubular cells. Arch Biochem Biophys 286: 46-56, 1991
Nelson TJ, Boor PJ: Allylamine cardiotoxicity: IV: Metabolism to acrolein by cardiovascular tissues. Biochem Pharmacol 31: 509-514, 1982
Ramos KS, Thurlow CH: Comparative cytotoxic responses of cultured avian and rodent aortic smooth muscle cells to allylamine. J Toxicol Environ Health 40: 61-76, 1993
He NG, Awasthi S, Singhal SS, Trent MB, Boor PJ: The role of glutathione-S-transferase as a defense against reactive electrophile in the blood vessel wall. Toxicol Appl Pharmacol 152: 83-89, 1998
Wei MX, Tamiya T, Rhee RJ, Breakefield XO, Chiocca EA: Diffusable cytotoxic metabolites contribute to the in vitro bystander effect associated with the cyclophosphamide/cytochrome P450 2BI cancer gene therapy paradigm. Clin Cancer Res 1: 1171-1177, 1995
Uchida K, Kanematsu M, Morimitsu Y, Osawa T, Noguchi N, Niki E: Acrolein is a product of lipid peroxidation reaction. Formation of free acrolein and its conjugate with lysine residue in oxidized low-density lipoproteins. J Biol Chem 273: 16058-16066, 1998
Kondo M, Oya-Ito T, Kumagai T, Osawa T, Uchida K: Cyclopentenone prostaglandins as potential inducers of intracellular oxidative stress. J Biol Chem 276: 12076-12083, 2001
Esterbauer H, Schauer RJ, Zollner H: Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic Biol Med 11: 81-128, 1991
Dorr RT, Lagel K: Effect of sulfhydryl compounds and glutathione depletion on rat heart myocyte toxicity induced by 4-hydroperoxycyclophosphamide and acrolein in vitro. Chem Biol Interact 93: 117-128, 1994
Markesbery WR: Oxidative stress hypothesis in Alzheimer's disease. Free Radic Biol Med 23: 134-147, 1997
Calingasan NY, Uchida K, Gibson GE: Protein-bound acrolein: A novel marker of oxidative stress in Alzheimer's disease. J Neurochem 72: 751-756, 1999
Ucida K: Cellular response to bioactive lipid peroxidation products. Free Radic Res 37: 731-737, 2000
Ramos K, Grossman SL, Cox LR: Allylamine-induced vascular toxicity in vitro: Prevention by semicarbazide-sensitive amine oxidase inhibitors. Toxicol Appl Pharmacol 95: 61-71, 1988
Boor PJ: Allylamine cardiovascular toxicity V: Tissue distribution and toxicokinetics after oral administration. Toxicology 35: 167-177, 1985
Conklin DJ, Lanagford SD, Boor PJ: Contribution of serum and cellular semicarbazide-sensitive amine oxidase to amine metabolism and cardiovascular toxicity. Toxicol Sci 46: 386-392, 1998
Hormann VA, Moore DR, Rikans LE: Relative contributions of protein sulfhydryl loss and lipid peroxidation to allyl alcohol-induced cytotoxicity in isolated rat hepatocytes. Toxicol Appl Pharmacol 98: 375-384, 1989
Armugam N, Thanislass J, Ragunath K, Devaraj SN, Devaraj H: Acrolein-induced toxicity - defective mitochondrial function as a possible mechanism. Arch Environ Contam Toxicol 36: 373-376, 1999
Boor PJ, Hysmith RM: Allylamine cardiovascular toxicity. Toxicology 44: 129-144, 1987
Cox LR, Ramos K: Allylamine-induced phenotypic modulation of aortic smooth muscle cells. J Exp Pathol 71: 11-18, 1990
Awasthi S, Boor PJ: Semicarbazide protection from in vivo oxidant injury of vascular tissue by allylamine. Toxicol Lett 66: 157-163, 1993
Misra P, Srivastava SK, Singhal SS, Awasthi S, Awasthi YC, Boor PJ: Glutathione-S-transferase 8-8 is localized in smooth muscle cells of rat aorta and is induced in an experimental model of atherosclerosis. Toxicol Appl Pharmacol 133: 27-33, 1995
Berhne K, Mannervik B: Inactivation of the genotoxic aldehyde acrolein by human glutathione transferases of classes Alpha, Mu, and Pi. Mol Pharmacol 37: 251-254, 1990
Tsuchida S, Sato K: Glutathione transferase and cancer. Crit Rev Biochem Mol Biol 27: 337-384, 1992
Tappel AL: Selenium-glutathione peroxidase: Properties and synthesis. Curr Top Cell Regul 24: 87-97, 1984
Anderson MM, Hazen SL, Hsu FF, Heinecke JW: Human neutrophils employ the mycloperoxidase-hydrogen peroxide-chloride system to convert hydroxy-amino acids into glycoaldehyde, 2-hydroxypropanal, and acrolein. J Clin Invest 99: 424-432, 1997
Witz G: Biological interactions of alpha, beta-unsaturated aldehydes. Free Radic Biol Med 7: 333-349, 1989
Horton ND, Biswal SS, Corrigan LL, Bratta J, Kehrer JP: Acrolein causes inhibitor κB-independent decreases in nuclear factor κB activation in human lung adenocarcinoma (A549) cells. J Biol Chem 274: 9200-9206, 1999
Nakamura H, Nakamura K, Yodoi J: Redox regulation of cellular activation. Ann Rev Immunol 15: 351-369, 1997
Dalton TP, Shertzer HG, Puga A: Regulation of gene expression by reactive oxygen. Ann Rev Pharmacol Toxicol 39: 67-101, 1999
Widmann C, Gibson S, Jarpe MB, Johnson GL: Mitogen-activated protein kinase: Conservation of a three-kinase module from yeast to human. Physiol Rev 79: 143-180, 1999
Kyriakis JM, Avruch J: Sounding the alarm: Protein kinase cascades activated by stress and inflammation. J Biol Chem 271: 24313-24316, 1996
Gerthoffer WT, Yamboliev IA, Shesrer M, Pohl J, Haynes R, Dang S, Sato K, Sellers JR: Activation of MAP kinases and phosphorylation of caldesmon in canine colonic smooth muscle. J Physiol 495: 597-609, 1996
Hedges JC, Dechert MA, Yamboliev IA, Martin JL, Hickey E, Weber LA, Gerthoffer WT: A role for p38MAPK/HSP27 pathway in smooth muscle cell migration. J Biol Chem 274: 24211-24219, 1999
Seger R, Krebs EG: The MAPK signaling cascade. FASEB J 9: 726-735, 1995
Guyton KZ, Liu Y, Gorospe M, Xu Q, Holbrook NJ: Activation of mitogen-activated protein kinase by H2O2. J Biol Chem 271: 4138-4142, 1996
Viedt C, Soto U, Krieger-Brauer HI, Fei J, Elsing C, Kubler W, Kreuzer J: Differential activation of mitogen-activated protein kinases in smooth muscle cells by angiotensin II. Arteroscler Thromb Vasc Biol 20: 940-948, 2000
Ramos K, Cox LR: Primary cultures of rat aortic endothelial and smooth muscle cells: An in vitro model to study xenobiotic-induced vascular toxicity. In Vitro Cell Dev Biol 23: 288-296, 1987
Hamilton LL, Holian A: Effect of acrolein on human alveolar macrophage NF-kappa B activity. Am J Physiol 277: L550-L557, 1999
Ranganna K, Yatsu FM, Hayes BE, Milton SG, Jayakumar A: Butyrate inhibits proliferation-induced proliferating cell nuclear antigen expression (PCNA) in rat vascular smooth muscle cells. Mol Cell Biochem 205: 149-161, 2000
Ranganna K, Joshi T, Yatsu FM: Sodium butyrate inhibits plateletderived growth factor-induced proliferation of vascular smooth muscle cells. Atheroscler Thromb Vasc Biol 15: 2273-2283, 1995
Fridovich I: The biology of oxygen radicals. Science 201: 875-880, 1978
Sohal RS, Allen RG: Oxidative stress as a causal factor in differentiation and aging: A unifying hypothesis. Exp Gerontol 25: 499-522, 1990
Bankson DD, Kestin M, Rifai N: Role of free radicals in cancer and atherosclerosis. Clin Lab Med 13: 463-480, 1993
Bhatnagar A: Electrophysiological effects of 4-hydroxynonenal, an aldehydic product of lipid peroxidation on isolated rat ventricular myocytes. Circ Res 76: 293-304, 1995
Wolff SP: Diabetes mellitus and free radicals. Free radicals, transition metals and oxidative stress in the etiology of diabetes mellitus and complications. Br Med Bull 49: 642-652, 1993
Ziv I, Melamed E, Nardi N, Luria D, Achiron A, Offen D, Barzilai A: Dopamine induces apoptosis-like cell death in cultured chick sympathetic neurons - a possible novel pathogenetic mechanism in Parkinson's disease. Neurosci Lett 170: 136-140, 1994
Heldin CH: Dimerization of cell surface receptors in signal transduction. Cell 80: 213-223, 1995
Nakashima I, Pu MY, Nishizaki A, Rosil I, Ma L, Katano Y, Ohkusu K, Rahman SM, Isobe K, Hamaguchi M: Redox mechanism as alternative to ligand binding for receptor activation delivering disregulated cellular signals. J Immunol 152: 1064-1071, 1994
Suzuki YJ, Forman HJ, Sevanian A: Oxidants as stimulators of signal transduction. Free Radic Biol Med 22: 269-285, 1997
Nakamura K, Hori T, Sato N, Sugie K, Kawakami T, Yodoi J: Redox regulation of a src family protein tyrosine kinase p56 (lck) in T-cells. Oncogene 8: 3133-3139, 1993
Sachsenmaier C, Radler-Pohl A, Zinck R, Nordheim A, Herrlich P, Rahmsdorf HJ: Involvement of growth factor receptors in the mammalian UVC response. Cell 78: 963-972, 1994
Fialkow L, Chan CK, Rotin D, Grinstein S, Downey GP: Activation of the mitogen-activated protein kinase signaling pathway in neutrophils. Role of oxidants. J Biol Chem 269: 31234-31242, 1994
Knebel A, Rahmsdorf HJ, Ullrich A, Herrlich P: Dephosphorylation of receptor tyrosine kinases as target of regulation by radiation, oxidants or alkylating agents. EMBO J 15: 5314-5325, 1996
Cavigelli M, Li W, Lin A, Su B, Yoshioka K, Karin M: The tumor promoter arsenite stimulates AP-1 activity by inhibiting a JNK phosphatase. EMBO J 15: 6296-6279, 1996
Yamagishi S, Yamada M, Ishikawa Y, Matsumoto T, Ikeuchi T, Hatanaka H: p38 Mitogen-activated protein kinase regulates low potassium-induced c-jun phosphorylation and apoptosis in cultured cerebellar granule neurons. J Biol Chem 276: 5129-5133, 2000
Dudley DT, Pang L, Decker SJ, Bridges AJ, Saltiel AR: A synthetic inhibitor of the mitogen-activated protein kinase cascade. Proc Natl Acad Sci USA 92: 7686-7689, 1995
Han Z, Boyle DL, Aupperle KR, Bennett B, Manning AM, Firestein GS: Jun N-terminal kinase in rheumatoid arthritis. J Pharmacol Exp Ther 291: 124-130, 1999
Cho S-G, Lee YH, Park H-S, Ryoo K, Kang KW, Park J, Eom S-J Kim MJ, Chang T-S, Choi S-Y, Shim J, Kim Y, Dong M-S, Lee M-J, Kim SG, Ichijo H, Choi E-J: Glutathione s-transferase Mu modulates the stress-activated signals by suppressing apoptosis signal-regulating kinase 1. J Biol Chem 276: 12749-12755, 2001
Adler V, Yin Z, Fuchs SY, Benezra M, Rosario L, Tew KD, Pincus MR, Sardana M, Henderson CJ, Wolf CR, Davis RJ, Ronai Z: Regulation of JNK signaling by GSTp. EMBO J 18: 1321-1334, 1999
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ranganna, K., Yousefipour, Z., Nasif, R. et al. Acrolein activates mitogen-activated protein kinase signal transduction pathways in rat vascular smooth muscle cells. Mol Cell Biochem 240, 83–98 (2002). https://doi.org/10.1023/A:1020659808981
Issue Date:
DOI: https://doi.org/10.1023/A:1020659808981