Archives of Toxicology

, Volume 93, Issue 10, pp 2927–2942 | Cite as

Protective effect of methylene blue on TNBS-induced colitis in rats mediated through the modulation of inflammatory and apoptotic signalling pathways

  • Nesrine S. El SayedEmail author
  • Aya S. Sayed
Organ Toxicity and Mechanisms


Ulcerative colitis (UC) is a common type of chronic, idiopathic inflammatory bowel disease (IBD) that affects the mucosal lining of the colon. Long-term UC remission has shed light on the necessity of modified therapeutic strategies. In this study, UC was induced in rats by intrarectal administration of 2,4,6-trinitrobenzene sulfonic acid (TNBS). The anticolitis effect of methylene blue (MB), a well-known dye and antioxidant and a potent mitochondrial enhancer was tested. MB was injected intraperitoneally (i.p.) at a dose of 1 mg/kg, 2 mg/kg or 4 mg/kg, and colosalazine was administered orally (p.o.) at a dose of 500 mg/kg 11 days after the administration of TNBS, which was injected on the 8th day. All treatment group results were compared to the TNBS group results. Macroscopically, limited body weight loss and decrease in the colon weight per unit length ratio were observed in the MB groups. MB improved histological damage and decreased the expression of myeloperoxidase (MPO) and accumulation of CD4+ lymphocytes observed by immunohistochemistry. Downregulation of Bax/Bcl2 protein expression was detected using Western blotting, and increased mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) was measured by qPCR. MB produced biochemical alterations, such as significant decrease in interleukin-6 (IL-6), interleukin-17 (IL-17) and intercellular adhesion molecule-1 (ICAM-1) levels measured by enzyme-linked immunosorbent assay (ELISA). Significant decrease in malondialdehyde (MDA) and inducible nitric oxide synthase (iNOS) levels as well as significant increase in superoxide dismutase (SOD) and mitochondrial cytochrome c oxidase levels were observed with MB, and these effects were similar to those produced by colosalazine. Thus, MB altered disease pathogenesis and could be a promising and challenging therapeutic target for UC treatment.


Colitis Methylene blue Apoptosis Colosalazine Mitochondrial dysfunction TNBS 



Analysis of variance


Carboxymethyl cellulose




Enzyme-linked immunosorbent assay


Inducible nitric oxide synthase


Inflammatory bowel disease




Intercellular adhesion molecule-1




Methylene blue




Nuclear factor erythroid 2-related factor 2


Phosphate-buffered solution


Reactive oxygen species


Superoxide dismutase


T helper


2,4,6-Trinitrobenzene sulfonic acid


Ulcerative colitis



This research received no specific grant from any funding agency in the public or commercial.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Pharmacology and Toxicology, Faculty of PharmacyCairo UniversityCairoEgypt
  2. 2.Department of Pharmaceutical Biology, Faculty of Pharmacy and BiotechnologyGerman University in CairoCairoEgypt

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