Regulatory report
Aminoguanidine and curcumin attenuated tumor necrosis factor (TNF)-α-induced oxidative stress, colitis and hepatotoxicity in mice

https://doi.org/10.1016/j.intimp.2011.10.010Get rights and content

Abstract

The up regulation of gut mucosal cytokines such as tumor necrosis factor (TNF)-α and oxidative stress have been related to inflammatory bowel diseases (IBD) such as ulcerative colitis (UC) and Crohn's disease (CD).

This study investigated an immune-mediated model of colitis. TNF-α injected intraperitonally to mice induced a dose-dependent recruitment of neutrophils into abdominal mesentery. The leukocytes influx induced by TNF-α (10 μg kg 1 body weight) increased by 3 fold liver and colon damage scores. TNF-α-colitis was characterized by hemorrhagic edemas and crypt abscesses massively infiltrated by inflammatory cells, namely neutrophils. Moreover, TNF-α-toxicity resulted in liver steatosis and foci of necrosis infiltrated by Kupffer cells and neutrophils in parenchyma and around the centrilobular veins.

The involvement of oxidative stress was evaluated using aminoguanidine (AG) as selective inhibitor of inducible NO synthase (iNOS) and curcumin (Cur), the polyphenolic antioxidant of turmeric (Curcuma longa L.). TNF-α-toxicity led to significant increase in myeloperoxidase (MPO, an index of neutrophils infiltration), nitrites (stable nitric oxide metabolites) and malondialdehyde (MDA, a marker of lipid peroxides) levels and cell apoptosis in liver and colon. AG and Cur treatments significantly attenuated the hallmarks of oxidative stress, neutrophils influx and ROS-related cellular and histological damages, in TNF-α-treated mice.

Taken together, our results provide insights into the role of phagocytes-derived oxidants in TNF-α-colitis in mice. Cur and AG, by inhibiting neutrophils priming and iNOsynthase could be effective against oxidative bowel damages induced in IBD by imbalanced gut immune response.

Introduction

Ulcerative colitis (UC) and Crohn's disease (CD) are the two most common chronic relapsing disorders of inflammatory bowel disease (IBD). The chronic bowel inflammation is driven by complex crosstalk between imbalanced immune response [1], [2] and oxidative stress [3], [4], [5], [6], [7]. Among the various gut immune factors, tumor necrosis factor-α (TNF-α) is a key pro-inflammatory cytokine, in IBD [1], [2], [8], [9]. Mucosal levels of TNF-α are elevated in IBD patients [10], [11] and their inhibition [12] or neutralization by anti-TNF-α therapies improved both UC [13] and CD [14]. Moreover, endogenous TNF-α binds to cell surface receptors TNF-R1 (p55) and TNF-R2 (p75) [15] and primes neutrophils, which massively accumulated into inflamed tissues [16]. The molecular mechanism involves the phosphorylation on Ser345-p47phox, the major cytosolic component of the phagocyte NADPH oxidase complex (NOX2) [17]. Primed neutrophils release high amounts of reactive oxygen species (ROS) in synergy with various granular bactericidal peptides and oxidative enzymes, such as myeloperoxidase (MPO), which not only enhance bactericidal efficiency but also drastically, contribute to oxidative stress-induced injuries [16], [17], [18].

Nitric oxide (NO) participates in various physio-pathological events in gastrointestinal tract. Physiological levels of NO released by the constitutive NOsynthase (cNOS) protect the gut mucosa, through the regulation of mucus secretion [18], [19]. However, the large amounts of NO generated by inducible NOsynthase (iNOS) is converted into the highly cytotoxic peroxynitrite, upon reaction with superoxide anion [20], [21], [22], [23]. In addition, the endothelial NOsynthase (eNOS) might be uncoupled by peroxynitrite and converted to a dysfunctional superoxide anion-releasing enzyme, thereby contributing to oxidative stress [24].

Numerous inflammatory diseases including IBD are multifactorial disorders with no therapeutic agents available to simultaneously control both inflammatory immune response and oxidative stress. Aminoguanidine (AG) is a selective inhibitor of iNOsynthase. It reduces the availability of NO by competing with the natural substrate l-arginine [25]. AG also acts as an antioxidant by scavenging ROS [26]. The protective effect of curcumin (Cur), a polyphenolic pigment of turmeric (Curcuma longa L.) on various oxidants-driven diseases is related to its potent ROS scavenging activity [27]. Moreover, Cur can modulate immune response through the inhibition of the key oxidative stress sensitive nuclear transcription factor kappaB (NF-κB) [28], thus resulting in the control of pro-inflammatory enzymes, such as iNOS. Cur also up regulates antioxidant genes expression such as nuclear factor (erythroid-derived 2)-related factor (NRF2) and phase II enzymes of xenobiotic metabolism [29].

Thus, the aim of this study was to evaluate the efficacy of aminoguanidine (AG), as selective iNOS inhibitor and Curcumin (Cur), a naturally occurring antioxidant, on TNF-α-induced oxidative stress, acute colitis and liver toxicity. This oxidative inflammation was assessed macroscopically, on histological and cellular changes and on oxidative stress markers.

Section snippets

Materials

Tumor necrosis factor (TNF-α) was from PeroTech EC (London, GB). Aminoguanidine, curcumin, o-dianisidine hydrochloride, N-(1-napthyl)-ethylenediamine dihydro-chloride, sulphanilamide and thiobarbituric acid were obtained from Sigma Aldrich (St Louis MO, USA). All other chemicals were of analytic grade.

Animals care

Male NMRI mice weighing 20–22 g (Institut Pasteur, Algiers, Algeria), were housed five per cage under controlled conditions throughout the experiments. Standard rodent chow and water were supplied

Dose dependency of TNF-α-induced acute colitis

Mice treated with saline, AG or Cur showed normal colon histology (Fig. 1; Table 3). In contrast, TNF-α induced a dose–dependent increase in colon damage scores. The macroscopic alterations index (1.6 ± 0.24, 2.6 ± 0.24, and 3.2 ± 0.37) reflected in histological lesions (1.2 ± 0.37, 2.2 ± 0.49 and 3.2 ± 0.37) induced by 1, 5 and 10 μg kg 1 TNF-α challenge, respectively (Table 3, Table 4). Wide hemorrhagic edemas and crypt abscesses massively infiltrated by neutrophils characterized the colons of mice

Discussion

Tumor necrosis factor (TNF)-α is a pro-inflammatory cytokine which plays a crucial role in the pathogenesis of IBD [1], [2], [8], [9]. Moreover, the imbalance between ROS and ROS-neutralizing systems was shown to be a potentiating etiological and/or triggering factor in this disease [3], [4], [5], [6], [7].

This study was conducted in mice, using an immune-mediated model of colitis. Our results showed that TNF-α-induced acute inflammation was reflected in the massive neutrophils influx,

Acknowledgments

The authors declare no conflict of interests. This work was supported by the “Programme National de Recherche en Sciences Fondamentales 2011”, Algiers, Algeria. We are grateful to Dr MC Dang from the “Centre de Recherche Biomédicale Bichat-Beaujon, CRB3”, Paris, France, for her kind gift of TNF-α.

References (58)

  • E. Stroes et al.

    Origin of superoxide production by endothelial nitric oxide synthase

    FEBS Lett

    (1998)
  • A.J. Ruby et al.

    Anti-tumour and antioxidant activity of natural Curcuminoids

    Cancer Lett

    (1995)
  • S. Singh et al.

    Activation of transcription factor NF-kB is suppressed by Curcumin (diferuloylmethane)

    J Biol Chem

    (1995)
  • R.N. Fedorak et al.

    Verapamil alters eicosanoid synthesis and accelerates healing during experimental colitis in rats

    Gastroenterology

    (1992)
  • J.E. Krawisz et al.

    Quantification assay for acute intestinal inflammation based on myeloperoxidase activity

    Gastroenterology

    (1984)
  • L.C. Green et al.

    Analysis of nitrate, nitrite and [15 N] nitrate in biological fluids

    Anal Biochem

    (1982)
  • H. Ohkawa et al.

    Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction

    Anal Biochem

    (1979)
  • S. Suzuki et al.

    Role of Kupffer cells and the spleen in modulation of endotoxin-induced liver injury after partial hepatectomy

    Hepatology

    (1996)
  • P. Romagnani et al.

    T cells and cytokines in Crohn's disease

    Curr Opin Immunol

    (1997)
  • B. Fromenty et al.

    Inhibition of mitochondrial beta-oxidation as a mechanism of hepatotoxicity

    Pharmacol Ther

    (1995)
  • D.P. Jones

    Disruption of mitochondrial redox circuitry in oxidative stress

    Chem Biol Interact

    (2006)
  • L. Fialkow et al.

    Reactive oxygen and nitrogen species as signalling molecules regulating neutrophil function

    Free Radic Biol Med

    (2007)
  • K. Nishida et al.

    Contribution of NO synthases to neutrophils infiltration in the gastric mucosal lesions in rats water immersion restraint stress

    FEBS Lett

    (1998)
  • F. Balkwill et al.

    Smoldering and polarized inflammation in the initiation and promotion of malignant disease

    Cancer Cell

    (2005)
  • P.W. Szlosarek et al.

    Tumour necrosis factor alpha: a potential target for the therapy of solid tumours

    Lancet Oncol

    (2003)
  • P. Szlosarek et al.

    Tumour necrosis factor-alpha as a tumour promoter

    Eur J Cancer

    (2006)
  • M.Y. Chan et al.

    In vivo inhibition of nitric oxide synthase gene expression by Curcumin, a cancer preventive natural product with anti-inflammatory properties

    Biochem Pharmacol

    (1998)
  • S.J. Brown et al.

    The immune response in inflammatory bowel disease

    Am J Gastroenterol

    (2007)
  • A. Keshavarzian et al.

    Role of reactive oxygen metabolites in experimental colitis

    Gut

    (1990)
  • Cited by (79)

    • Nano-curcumin therapy, a promising method in modulating inflammatory cytokines in COVID-19 patients

      2020, International Immunopharmacology
      Citation Excerpt :

      Curcumin, as a prominent immune modulatory factor, suppresses the expression of multiple pro-inflammatory cytokines, such as IL-6, IL-10, TNF-α, IL-12, and chemokines. According to several studies, curcumin is able to decrease the mRNA expression and cytokine secretion level of IL-1, IL-6 and TNF-α in inflammatory disease, as discussed in the study of Yetkin et al. [34], Schneider et al. [35] and Mouzaoui et al. [36]. A meta-analysis by Derosa et al. also revealed that curcumin was able to decrease the amount of circulating IL-6 in patients with systemic inflammation [37], while the same effect of curcumin on TNF-α was observed in meta-analysis of Sahebkar et al. [38].

    • Nutraceuticals and biotics in pediatric gastrointestinal disorders

      2024, European Journal of Clinical Nutrition
    View all citing articles on Scopus
    View full text