Anat Cell Biol.  2010 Mar;43(1):44-53. 10.5115/acb.2010.43.1.44.

Recovered changes in the spleen by agmatine treatment after transient cerebral ischemia

• Uranchimeg D
• Kim JH
• Kim JY
• Lee WT
• Park KA
• Batbaatar
• Tundevrentsen S
• Amgalanbaatar D
• Lee JE

• KMID: 2168896
• DOI: http://doi.org/10.5115/acb.2010.43.1.44

Abstract

Stroke or cerebrovascular injury is the leading cause of disability and the third leading cause of deaths worldwide. After the initial ischemic injury, sympathetic signals are transmitted to the spleen and a compromised blood-brain barrier, coupled with expression of adhesion molecules by the vascular endothelial cells permits an influx of peripheral immune cells. This influx of peripheral immune cells into the brain exacerbates the local brain inflammatory response, leading to enhanced neurodegeneration. Agmatine is a primary amine formed by decarboxylation of L-arginine synthesized in the mammalian brain. In this study, we determined the effect of agmatine on the immune response in the spleen after transient cerebral ischemia. Twenty-three hours after transient cerebral ischemia, the white pulp area was reduced and the number of CD11b+ macrophages and CD4+CD25+ regulatory T cells (T reg cells) were increased in the spleens in the experimental group as a result of alteration of the immune response in the spleen, as regulated by inflammatory cytokines. In the agmatine treatment group (100 mg/kg IP), the contraction of white pulp was diminished and the number of CD11b+ macrophages and CD4+CD25+T reg cells were decreased. Twenty-three hours after transient cerebral ischemia, the brain infarction area was significantly reduced (5.51+/-1.63% of the whole brain) in the agmatine treatment group compared to 15.02+/-4.28% of the whole brain in the experimental control group. These results suggest that agmatine treatment can reduce brain infarction through minimizing neuroinflammation and can lessen the danger of post-stroke infection from depression of the immune system after stroke.

Keyword

Spleen; agmatine; macrophages; regulatory T (T reg) cells; cerebral ischemia

MeSH Terms

Agmatine
Arginine
Blood-Brain Barrier
Brain
Brain Infarction
Brain Ischemia
Cause of Death
Contracts
Cytokines
Decarboxylation
Depression
Endothelial Cells
Humans
Immune System
Ischemic Attack, Transient
Macrophages
Spleen
Stroke
T-Lymphocytes, Regulatory
Agmatine
Arginine
Cytokines

Figure

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  Fig. 1 TTC staining of the ischemic injured brain with or without agmatine treatment. Agmatine treatment reduced the infarction (white color area) 23 hours after transient cerebral ischemia compared to experimental control. NC, Normal control group; EC, Experimental control group; AGM, Agmatine treatment group.

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  Fig. 2 Macrograph of spleen tissue stained with hematoxylin-eosin. Red pulp and white pulp map was clearly distinguished and the number of white pulp was increased in agmatine treatment group compared to experimental control group. Scale bar is 200 µm. NC, Normal control group; EC, Experimental control group; AGM, Agmatine treatment group.

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  Fig. 3 The percentage of red and white pulp areas of the spleen 23 hours after transient cerebral ischemia. Agmatine treatment significantly increased the white pulp area and decreased the red pulp area 23 hours after transient cerebral ischemia compared to experimental control group. NC, Normal control group; EC, Experimental control group; AGM, Agmatine treatment group (*P<0.05 vs EC; †P<0.05 vs NC and AGM).

• 

  Fig. 4 The change of CD11b+ macrophages in the spleen 23 hours after transient cerebral ischemia. (A) Macrograph of CD11b+ macrophages in the spleen 23 hours after transient cerebral ischemia. Agmatine treatment reduced CD11b+ macrophages (brown) in the white pulp compared to the experimental control group. (B) in the experimental control group, the total number of CD11b+ macrophages was increased in the white pulp of whole spleen compared to the normal control group, but the total number of CD11b+ macrophages was decreased in the agmatine treatment group compared to the experimental control group. (C) The number of CD11b+ macrophages per unit volume in the white pulp of the spleen was the highest in the experimental control group and it was significantly decreased in the agmatine treatment group compared to the normal control group. Scale bar is 50 µm. NC, Normal control group; EC, Experimental control group; AGM, Agmatine treatment group (*P<0.05 vs EC).

• 

  Fig. 5 The change of CD4+CD25+ T reg cells in the spleen 23 hours after transient cerebral ischemia. (A) Macrograph of CD4+CD25+ T reg cells in the spleen 23 hours after transient cerebral ischemia. Agmatine treatment reduced CD4+CD25+ T reg cells (purple) in the white pulp compared to experimental control group. (B) In the experimental control group, the total number of CD4+CD25+ T reg cells was increased in the white pulp of the entire spleen compared to the normal control group, but the total number of CD4+CD25+ T reg cells was decreased in the agmatine treatment group compared to the experimental control group. (C) The number of CD4+CD25+ T reg cells per unit volume in the white pulp of the spleen was the highest in the experimental control group and it was significantly decreased in the agmatine treatment group as the normal control group. Scale bar is 50 µm. NC, Normal control group; EC, Experimental control group; AGM, Agmatine treatment group (*P<0.05 vs EC).

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  Fig. 6 The change of CD8+ CTLs in the spleen 23 hours after transient cerebral ischemia. (A) Macrograph of CD8+ CTLs in the spleen 23 hours after transient cerebral ischemia. CD8+ CTLs (brown) were increased in the experimental control and agmatine treatment groups compared to the normal control group. (B) in the experimental control group, the total number of CD8+ CTLs was increased in the white pulp of the entire spleen compared to the normal control group, but it was not significant. The total number of CD8+ CTLs in the agmatine treatment group was increased without significance. (C) The number of CD8+ CTLs per unit volume in the white pulp of the spleen was the highest in the experimental control group, but it was not significant among all groups. Scale bar is 50 µm. NC, Normal control group; EC, Experimental control group; AGM, Agmatine treatment group.

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