Anat Cell Biol.  2018 Dec;51(4):292-298. 10.5115/acb.2018.51.4.292.

Amelioration of experimental autoimmune encephalomyelitis by Ishige okamurae

• Ahn M
• Kim J
• Yang W
• Choi Y
• Ekanayake
• Ko H
• Jee Y
• Shin T

• KMID: 2430197
• DOI: http://doi.org/10.5115/acb.2018.51.4.292

Abstract

Experimental autoimmune encephalomyelitis (EAE) is a T-cell-mediated autoimmune central nervous system disease characterized by inflammation with oxidative stress. The aim of this study was to evaluate an anti-inflammatory effect of Ishige okamurae on EAE-induced paralysis in rats. An ethanolic extract of I. okamurae significantly delayed the first onset and reduced the duration and severity of hind-limb paralysis. The neuropathological and immunohistochemical findings in the spinal cord were in agreement with these clinical results. T-cell proliferation assay revealed that the ethyl-acetate fraction of I. okamurae suppressed the proliferation of myelin basic protein reactive T cells from EAE affected rats. Flow cytometric analysis showed TCRαβ+ T cells was significantly reduced in the spleen of EAE rats with I. okamurae treatment with concurrent decrease of inflammatory mediators including tumor necrosis factor-α and cyclooxygenase-2. Collectively, it is postulated that I. okamurae ameliorates EAE paralysis with suppression of T-cell proliferation as well as decrease of pro-inflammatory mediators as far as rat EAE is concerned.

Keyword

Experimental autoimmune encephalomyelitis; Inflammation; Ishige okamurae; Spinal cord

MeSH Terms

Animals
Central Nervous System
Cyclooxygenase 2
Encephalomyelitis, Autoimmune, Experimental*
Ethanol
Inflammation
Myelin Basic Protein
Necrosis
Oxidative Stress
Paralysis
Rats
Spinal Cord
Spleen
T-Lymphocytes
Cyclooxygenase 2
Ethanol
Myelin Basic Protein

Figure

• 

  Fig. 1 Histopathological examination of the spinal cords of rats with experimental autoimmune encephalomyelitis (EAE) with or without Ishige okamurae treatment. There was no infiltration of inflammatory cells in the spinal cord of normal control rats (A, D). The spinal cords of vehicle-treated EAE rats contained many inflammatory cells (arrowheads) (B, E) in the parenchyma, whereas the number of inflammatory cells (arrowheads) (C, F) in the spinal cords of I. okamurae-treated rats (10 mg/kg body weight) was reduced. Hematoxylin and eosin staining. Arrowheads indicate inflammatory cells. Scale bars=200 µm (A–C) and 50 µm (D–F).

• 

  Fig. 2 Representative photographs of ionized calcium-binding adaptor molecule 1 (Iba-1) in the spinal cords of normal control (A), vehicle-treated (B), and Ishige okamurae-treated (C) experimental autoimmune encephalomyelitis (EAE) rats. (D) The Iba-1-positive area in the spinal cord of I. okamurae-treated EAE rats was significantly reduced compared with that of vehicle-treated EAE rats. *P<0.05. Insets indicate that higher-magnification photos. Immunostained for Iba-1 and counterstained with hematoxylin. Scale bars=200 µm (A–C), 50 µm (insets).

• 

  Fig. 3 Effect of Ishige okamurae on myelin basic protein-reactive T-cell responses (n=3). Data are means±SEM. CPM, count per minute; EAE, experimental autoimmune encephalomyelitis. ***P<0.001 vs. vehicle-treated rats.

• 

  Fig. 4 Ishige okamurae altered the population of mononuclear cells (MNCs) in spleen. (A) Splenic MNCs were isolated on day 10 postimmunization from rats treated with either vehicle or I. okamurae and analyzed by flow cytometry for TCRαβ+ T cells. The dot plot represents one of three separate experiments of similar observation. (B) Mean number of cell populations from splenic MNCs are plotted (n=3 each per group). Data are means±SEM. MNC, mononuclear cell. *P<0.05 vs. vehicle-treated rats.

• 

  Fig. 5 Quantitative real time polymerase chain reaction results of in the spinal cords of experimental autoimmune encephalomyelitis (EAE)-induced rats with or without Ishige okamurae at peak stage (n=3 per group). I. okamurae suppressed the pro-inflammatory mediator related genes in EAE-induced rats. The mRNA expression levels of tumor necrosis factor-α (TNF-α) (A) and cyclooxygenase-2 (COX-2) (B) in the spinal cords of I. okamurae-treated EAE rats was significantly decreased compared with that of vehicle-treated EAE rats. *P<0.05 vs. vehicle treated EAE rats.

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