Drug Res (Stuttg) 2015; 65(09): 495-499
DOI: 10.1055/s-0034-1389950
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

Plumbagin Protects Against Spinal Cord Injury-induced Oxidative Stress and Inflammation in Wistar Rats through Nrf-2 Upregulation

W. Zhang
1   Department of Orthopedics, Qilu Hospital, Shandong University, Jinan, Shandong, China
,
L. Cheng
1   Department of Orthopedics, Qilu Hospital, Shandong University, Jinan, Shandong, China
,
Y. Hou
1   Department of Orthopedics, Qilu Hospital, Shandong University, Jinan, Shandong, China
,
M. Si
1   Department of Orthopedics, Qilu Hospital, Shandong University, Jinan, Shandong, China
,
Y.-P. Zhao
1   Department of Orthopedics, Qilu Hospital, Shandong University, Jinan, Shandong, China
,
L. Nie
1   Department of Orthopedics, Qilu Hospital, Shandong University, Jinan, Shandong, China
› Author Affiliations
Further Information

Publication History

received 04 August 2014

accepted 21 August 2014

Publication Date:
22 September 2014 (online)

Abstract

Background: Spinal cord injury causes post-traumatic degeneration through series of biochemical events. This study aims to evaluate the possible protective mechanism of Plumbagin against Spinal cord injury induced oxidative stress and inflammation. Plumbagin is a potent antioxidant and shows anti-carcinogenic, anti-inflammatory and analgesic activities. However, its exact molecular mechanism of action has yet to be explored.

Methods: We tested the effects of Plumbagin on spinal cord injury induced ROS generation and lipid peroxidation content in wistar rats. Additionally, the expression of 2 important transcription factors NF-κB and Nrf-2 was investigated.

Results: Plumbagin treatment significantly ameliorated oxidative stress through inhibition of ROS and lipid peroxidation with a concomitant increase in antioxidant status. Western blot analysis revealed enhanced nuclear levels of Nrf-2, while NF-κB expression was suppressed during Plumbagin administration. Enzyme linked immunosorbent assay for pro-inflammatory cytokines (TNF-α, IL-1β) showed a significant downregulation followed by Plumbagin treatment in spinal cord injury rats.

Conclusion: Taken together, the data suggests potential and novel role of Plumbagin in cytoprotection by modulating NF-κB and Nrf-2 levels against spinal cord injury.

 
  • References

  • 1 Zhang N, Yin Y, Xu SJ et al. Inflammation & apoptosis in spinal cord injury. Indian J Med Res 2012; 135: 287-296
  • 2 Liu NK, Zhang YP, Titsworth WL et al. A novel role of phospholipase A2 in mediating spinal cord secondary injury. Ann Neurol 2006; 59: 606-619
  • 3 Hall ED, Springer JE. Neuroprotection and acute spinal cord injury: a reappraisal. Neuro Rx 2004; 1: 80-100
  • 4 Prasad VS, Devi PU, Rao BS et al. Radiosensitizing effect of plumbagin on mouse melanoma cells grown in vitro. Indian J Exp Biol 1996; 34: 857-858
  • 5 Singh UV, Udupa N. Reduced toxicity and enhanced antitumor efficacy of cyclodextrin plumbagin inclusion complex in mice bearing Ehrlich ascites carcinoma. Indian J Physiol Pharmacol 1997; 41: 171-175
  • 6 Devi PU, Rao BS, Solomon FE. Effect of plumbagin on the radiation induced cytogenetic and cell cycle changes in mouse Ehrlich ascites carcinoma in vivo. Indian J Exp Biol 1998; 36: 891-895
  • 7 Sugie S, Okamoto K, Rahman KM et al. Inhibitory effects of plumbagin and juglone on azoxymethane-induced intestinal carcinogenesis in rats. Cancer Lett 1998; 127: 177-183
  • 8 Wang CC, Chiang YM, Sung SC et al. Plumbagin induces cell cycle arrest and apoptosis through reactive oxygen species/c-Jun N-terminal kinase pathways in human melanoma A375.S2 cells. Cancer Lett 2008; 259: 82-98
  • 9 Luo P, Wong YF, Ge L et al. Anti-inflammatory and analgesic effect of plumbagin through inhibition of nuclear factor-kB activation. J Pharmacol Exp Ther 2010; 335: 735-742
  • 10 Erşahın M, Toklu HZ, Erzık C et al. Ghrelin alleviates spinal cord injury in rats via its anti-inflammatory effects. Turk Neurosurg 2011; 21: 599-605
  • 11 Allen AR. Surgery of experimental lesion of spinal cord equivalent to crush injury of fracture dislocation of spinal column: a preliminary report. JAMA 1911; 57: 878-880
  • 12 Lowry OH, Rosenbrough NJ, Farr AL et al. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193: 265-275
  • 13 Hillegas LM, Griswold DE, Brickson B et al. Assessment of myeloperoxidase activity in whole rat kidney. J Pharmacol Methods 1990; 24: 285-295
  • 14 Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbitturic acid reaction. Anal Biochem 1979; 95: 351-358
  • 15 Beutler E. Glutathione in red blood cell metabolism. In: Beutler (ed.). Red Cell Metabolism A manual of biochemical methods. Grune & Stratton; New York: 1975: 112-114
  • 16 Lind C, Cadenas E, Hochstein P et al. DT-diaphorase: Purification, ­properties, and function. Methods Enzymol 1990; 186: 287-301
  • 17 Aebi H. Catalase in vitro Methods Enzymol 1984; 105: 1-126
  • 18 Sun Y, Oberley LW, Ying L. A simple method for clinical assay of superoxide dismutase. Clin Chemc 1988; 34: 497-500
  • 19 Paglia DE, Valentine WN. Studies on the quantitative andqualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 1967; 70: 158-169
  • 20 Habig WH, Pabst MJ, Jakoby WB. GlutathioneS-transferases The first enzymatic step in mercapturic acid formation. J Biol Chem 1974; 249: 7130-7139
  • 21 Kwon S, Newcomb RL, George SC. Mechanisms of synergistic cytokine-induced nitric oxide production in human alveolar epithelial cells. Nitric Oxide 2001; 5: 534-546
  • 22 Kawamori D, Kajimoto Y, Kaneto H et al. Oxidative stress induces nucleo-cytoplasmic translocation of pancreatic transcription factor PDX-1 through activation of c-Jun NH(2)-terminal kinase. Diabetes 2003; 52: 2896-2904
  • 23 Rasband WS Image J. U. S. National Institutes of Health, Bethesda, Maryland, USA http://imagej.nih.gov/ij/ 1997–2014
  • 24 Parola M, Bellomo G, Robino G et al. 4-Hydroxynonenal as a biological signal: molecular basis and pathophysiological implications. Antioxid Redox Signal 1999; 1: 255-284
  • 25 Wenge Li, Tin OoKhor, Changjiang Xu et al. Activation of Nrf2-antioxidant signaling attenuates NF-κB-inflammatory response and elicits apoptosis. Biochem Pharmacol 2008; 76: 1485-1489
  • 26 Chen F, Castranova V, Shi X et al. New insights into the role of nuclear factor-kappaB, a ubiquitous transcription factor in the initiation of diseases. Clin Chem 1999; 45: 7-17
  • 27 Mercurio F, Manning AM. Multiple signals converging on NF-kappaB. Curr Opin Cell Biol 1999; 11: 226-232
  • 28 Rafati DS, Geissler K, Johnson K et al. Nuclear factor-kappaB decoy amelioration of spinal cord injury-induced inflammation and behavior outcomes. J Neurosci Res 2008; 86: 566-580
  • 29 Li Z, Xiao J, Wu X et al. Plumbagin inhibits breast tumor bone metastasis and osteolysis by modulating the tumor-bone microenvironment. Curr Mol Med 2012; 12: 967-981
  • 30 Wang T, Wu F, Jin Z et al. Plumbagin inhibits LPS-induced inflammation through the inactivation of the nuclear factor-kappa B and mitogen activated protein kinase signaling pathways in RAW 2647 cells. Food Chem Toxicol 2014; 64: 177-183
  • 31 Sandur SK, Ichikawa H, Sethi G et al. Plumbagin (5-hydroxy-2-­methyl-1, 4-naphthoquinone) suppresses NF-kappaB activation and NF-kappaB-regulated gene products through modulation of p65 and IkappaBalpha kinase activation, leading to potentiation of apoptosis induced by cytokine and chemotherapeutic agents. J Biol Chem 2006; 281: 17023-17033
  • 32 Checker R, Sharma D, Sandur SK et al. Anti-inflammatory effects of plumbagin are mediated by inhibition of NF-kappaB activation in lymphocytes. Int Immunopharmacol 2009; 9: 949-953
  • 33 Surh YJ. NF-kappa B and Nrf2 as potential chemopreventive targets of some anti-inflammatory and antioxidative phytonutrients with anti-inflammatory and anti-oxidative activities. Asia Pac J Clin Nutr 2008; 17: 269-272
  • 34 Son TG, Camandola S, Arumugam TV et al. Plumbagin, a novel Nrf2/ARE activator, protects against cerebral ischemia. J Neuro chem 2010; 112: 1316-1326
  • 35 Karuri AR, Huang Y, Bodreddigari S et al. 3H-1,2-Dithiole-3-thione targets nuclear factor κB to block expression of inducible nitric oxide synthase, prevents hypotension, and improves survival in endotoxemic rats. J Pharmacol Exp Ther 2006; 317: 61-67
  • 36 Liu GH, Qu J, Shen X. NF-kappaB/p65 antagonizes Nrf2-ARE pathway by depriving CBP from Nrf2 and facilitating recruitment of HDAC3 to MafK. Biochim Biophys Acta 2008; 1783: 713-727