Abstract
This study was designed to determine levels of NF-κB reporter gene activity and free radical generation in cultured striated myocytes (H9C2 cells) exposed to cocaine or morphine in the presence of free radical scavengers. Cells were transiently transfected with a NF-κB reporter gene and changes in luciferase activity were detected, by bioluminescence. Using confocal microscopy and 2′,7′-dichlorofluorescin diacetate, cocaine-induced or morphine-induced free radicals were quantified in H9C2 cells. Cocaine and morphine (0–1×10−2 M) were tested separately. Cocaine but not morphine significantly activated Nf-κB reporter gene, activity in H9C2 cells. Overexpression of IκB inhibited NF-κB reporter activity at low (1×10−4 M) but not high (1×10−2 M) cocaine concentrations. Free radicals were generated in H9C2 cells stimulated with cocaine but not with morphine. The production of free radicals and NF-κB reporter gene activity could be blocked with N-acetylcysteine, glutathione, and to a lesser extent, lipoic acid. The results suggest that cocaine induces free radical production, which leads to the activation of NF-κB signal transduction and possible inflammatory responses.
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Jaspers, I., Chen, L. C., and Flescher, E. (1998). Induction of interleukin-8 by ozone is mediated by tyrosine kinase and protein kinase A, but not by protein kinase C. J. Cell. Physiol. 177:313–323.
Ogata, N., Yamamoto, H., Kugiyama, K., Yasue, H., and Miyamoto, E. (2000). Involvement of protein kinase C in superoxide anion-induced activation of nuclear factor-kappa B in human endothelial cells. Cardiovasc. Res. 45:513–521.
Sen, R. and Baltimore, D. (1986). Multiple nuclear, factors interact with the immunoglobulin enhancer sequences. Cell 46:705–716.
Kaltschmidt, C., Kaltschmidt, B., Neumann, H., Wekerle, H., and Baeuerle, P.A. (1994). Constitutive NF-kappa B activity in neurons. Mol. Cell. Biol. 14:3981–3992.
Lawrence, R., Chang, L.J., Siebenlist, U., Bressler, P., and Sonenshein, G.E. (1994). Vascular smooth muscle cells express a constitutive NF-kappa B-like activity. J. Biol. Chem. 269:28913–28918.
Grimm, S. and Baeuerle, P.A. (1993). The inducible transcription factor NF-κB: Structure-function relationship of its protein subunits. J. Biochem. 290:297–308.
Baeuerle, P.A. and Baltimore, D. (1988). I kappa B: a specific inhibitor of the NF-kappa B transcription factor. Science 242:540–546.
Baeuerle, P.A. and Baltimore, D. (1988). Activation of DNA-binding activity in an apparently cytoplasmic precursor of the NF-B transcription factor. Cell 53:211–217.
Henkel, T., Machleidt, T., Alkalay, I., Kronke, M., Ben-Neriah, Y., and Baeuerle, P.A. (1993). Rapid proteolysis of I kappa B-alpha is necessary for activation of transcription factor NF-kappa B. Nature 365:182–185.
Palombella, V.J., Rando, O.J., Goldberg, A.L., and Maniatis, T. (1994). The ubiquitin-proteasome pathway is required for processing the NF-ekaoppa B1 precursor protein and the activation of NF-kappa B. Cell 78:773–785.
Traenckner, E.B., Pahl, H.L., Henkel, T., Schmidt, K.N., Wilk, S., and Baeuerle, P.A. (1995). Phosphorylation of human I kappa B-alpha on serines 32 and 36 controls I kappa B-alpha proteolysis and NF-kappa B activation in response to diverse stimuli. J. EMBO 14:2876–2883.
Thompson, J.E., Phillips, R.J., Erdjument-Bromage, H., Tempst, P., and Ghosh, S. (1995). I kappa B-beta regulates the persistent response in a biphasoc activation of NF-kappa B. Cell 80:573–582.
Grilli, M., Chiu, J.J., and Lenardo, M.J. (1993). NF-kappa B and Rel: participants in a multiform transcriptional regulatory system. Int. Rev. Cytol. 143:1–62.
Siebenlist, U., Franzoso, G., and Brown, K. (1994). Structure, regulation and function of NF-kappa B. Annu. Rev. Cell. Biol. 10:405–455.
Baeuerle, P.A. and Baltimore, D. (1996). NF-kappa B: ten years after. Cell 87:13–20.
Landry, D.B., Couper, L.L., Bryant, S.R., and Lindner., V. (1997). Activation of the NF-kappa B and I kappa B system in smooth muscle cells after rat arterial injury. Induction of vascular cel adhesion molecule-1 and, monocyte chemoattractant protein-1. Am. J. Pathol. 151:1085–1095.
Brach, M.A., Hass, R., Sherman, M.L., Gunji, H., Weichselbaum, R., and Kufe, D. (1991). Ionizing radiation induces expression and binding activity of the nuclear factor kappa B. J. Clin. Invest. 88:691–695.
Schreck, R., Rieber, P., and Baeuerle, P.A. (1991). Reactive oxygen intermediates as a pparently widely used messengers in the activation of the NF-kappa B transcription factor and HIV-1. J EMBO 10:2247–2258.
Peng, M., Huang, L., Xie, Z.J., Huang, W.H., and Askari, A. (1995). Oxidant-induced activations of nuclear factor-kappa B and activator protein-1 in cardiac myocytes. Cell. Mol. Biol. Res. 41:189–197.
Hattori, Y., Akimoto, K., Murakami, Y., and Kasai K. (1997). Pyrrolidine dithio carbamate inhibits cytokine-induced VCAM-1 gene expression in rat cardiac myocytes. Mol. Cell. Biochem. 177:177–181.
Kacimi, R., Karliner, J.S., Koudssi, F., and Long, C.S. (1998). Expression and regulation of adhesion molecules in cardiac cells by cytokines: response to acute hypoxia. Circ. Res. 82:576–586.
Li, C., Browder, W. and Kao, R.L. (1999). Early activation of transcription factor NF-kappa B during ischemia in perfused rat heart. Am. J. Physiol. 276:H543-H552.
Chandrasekar, B., Smith, J.B., and Freeman, G.L. (2001). Ischemia-reperfusion of rat myocardium activates nuclear factor-KappaB and induces neutrophil infiltration via lipopolysaccharide-induced CXC chemokine. Circulation 103: 2296–2302.
Altavilla, D., Deodato, B., Campo, G.M., Arlotta, M., Miano, M., Squadrito, G., et al. (2000). IRFI 042, a novel dual vitamin E-like antioxidant, inhibits activation of nuclear factor-kappaB and reduces the inflammatory response in myocardial ischemia-reperfusion injury. Cardiovasc. Res. 47:515–528.
Wong, S.C., Fukuchi, M., Melnyk, P., Rodger, I., and Giaid, A. (1998). Induction of cyclooxygenase-2 and activation of nuclear factor-kappaB in myocardium of patients with congestive heart failure. Circulation 98:100–103.
Hargrave, B.Y. and Lattanzio, F. (2002). Cocaine activates renin angiotensin system in pregnant rabbits and alters the response to ischemia. Cardiovasc. Toxicol. 2:91–97.
Yeh, S.Y., Gorodetzky, C.W., and Krebs, H.A. (1977). Isolation and identification of morphine 3- and 6-glucuronides, morphine 3,6-diglucuronide, morphine 3 ethereal sulfate, normorphine and normorphine 6-glucuronide as morphine metabolites in humans. J. Pharmacol. Sci. 66:1288–1293.
Oliveira, M.T., Rego, A.C., Morgadinho, M.T. Macedo, T.R., and Oliveira, C.R. (2002). Toxic effects of opioid and stimulant drugs on undifferentiated PC12 cells. Ann. N.Y. Acad. Sci. 965:487–496.
Yu, R.C.T., Lee, T.C., Wang, T.C., and Li, J.H. (1999). Genetic toxicity of cocaine. Carcinogenesis 20:1193–1199.
Cohen, M.V., Yang, X.M., Liu, G.S., Heusch, G., and Downey, J.M. (2001). Acetylcholine, bradykinin, opioids and phenylephrine, but not adenosine, trigger preconditioning by generating free radicals and opening mitochondrial Katp channels. Circ. Res. 89:273–278.
McPherson, B.C. and Yao, Z. (2001). Signal transduction of opioid-induced cardioprotection in ischemia-perfusion. Anesthesiology 94:1082–1088.
Fantel, A. and Person, R.E. (2002). Involvement of mitochondria and other free radical sources in normal and abnormal fetal development. Ann. N. Y. Acad. Sci. 959:424–433.
Castelli, M.C., Venturini, L., and Sparber, S.B. (2001). Cocaine and salicylate; documentation of hydroxyl radical formation in hearts and brains of 18-day-old chick embryos and unexpected interactive toxicity. Psychopharmacology 156:23–31.
Yang, Y., Xiao, H., Ke, Q., Cai, J., Xiao, Y.F., and Morgan, J.P. (2002). Enhancement of nitric oxide production by methylergonovine in cultured neonatal rat cardiomyocytes. Br. J. Pharmacol. 135:188–196.
Regnier, C.H., Song, H.Y., Gao, X., Goeddel, D.V., Cao, Z., and Rothe, M. (1997). Identification and characterization of an IkappaB kinase. Cell. 90:373–383.
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Hargrave, B.Y., Tiangco, D.A., Lattanzio, F.A. et al. Cocaine, not morphine, causes the generation of reactive oxygen species and activation of NF-κB in transiently cotransfected heart cells. Cardiovasc Toxicol 3, 141–151 (2003). https://doi.org/10.1385/CT:3:2:141
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DOI: https://doi.org/10.1385/CT:3:2:141