Korean J Physiol Pharmacol.  2010 Oct;14(5):257-263. 10.4196/kjpp.2010.14.5.257.

Neuroprotective Effect of Visnagin on Kainic Acid-induced Neuronal Cell Death in the Mice Hippocampus

• Kwon MS
• Lee JK
• Park SH
• Sim YB
• Jung JS
• Won MH
• Kim SM
• Suh HW

• KMID: 2071691
• DOI: http://doi.org/10.4196/kjpp.2010.14.5.257

Abstract

Visnagin (4-methoxy-7-methyl-5H-furo[3,2-g][1]-benzopyran-5-one), which is an active principle extracted from the fruits of Ammi visnaga, has been used as a treatment for low blood-pressure and blocked blood vessel contraction by inhibition of calcium influx into blood cells. However, the neuroprotective effect of visnagin was not clearly known until now. Thus, we investigated whether visnagin has a neuroprotective effect against kainic acid (KA)-induced neuronal cell death. In the cresyl violet staining, pre-treatment or post-treatment visnagin (100 mg/kg, p.o. or i.p.) showed a neuroprotective effect on KA (0.1 microgram) toxicity. KA-induced gliosis and proinflammatory marker (IL-1beta, TNF-alpha, IL-6, and COX-2) inductions were also suppressed by visnagin administration. These results suggest that visnagin has a neuroprotective effect in terms of suppressing KA-induced pathogenesis in the brain, and that these neuroprotective effects are associated with its anti-inflammatory effects.

Keyword

Neuroprotection; Kainic acid; Hippocampus; Visnagin; Cytokines

MeSH Terms

Ammi
Animals
Benzoxazines
Blood Cells
Blood Vessels
Brain
Calcium
Cell Death
Contracts
Cytokines
Fruit
Gliosis
Glycosaminoglycans
Hippocampus
Interleukin-6
Kainic Acid
Khellin
Mice
Neurons
Neuroprotective Agents
Tumor Necrosis Factor-alpha
Viola
Benzoxazines
Calcium
Cytokines
Glycosaminoglycans
Interleukin-6
Kainic Acid
Khellin
Neuroprotective Agents
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1. The effect of visnagin administered orally or intraperitoneally on KA-induced neuronal death in the hippocampus. The location of the hippocampal CA3 region performing cresyl violet-positive neuronal count is indicated (A) referring Franklin [18]. I.c.v. kainic acid injecton induced neuronal cell death in the pyramidal cells in the CA3 region of hippocampus (B). Mice were administered visnagin orally (C) or intraperitoneally (D) 20 min (– 20) prior to KA (0.1 μg/5 μl) injection or 0, 20 (+ 20), 40 (+40), 60 (+ 60) min after KA treatment (0.1 μg/5 μl). And then, the cresyl violet staining was performed at 1 day after KA. The vertical bars indicate the standard error of mean. ∗p<0.05, ∗∗p<0.01, ∗∗∗p<0.001 (KA 0.1 vs other groups). The mice number of each group was 10.

• 

  Fig. 2. The time course alteration of proinflammatory cytokines mRNA on KA injection in the hippocampus. Alteration of IL-1β, TNF-α (B), IL-6 (C), and COX-2 (D) mRNA at 1, 3, 6, 12, 24 hrs after intracerebral (i.c.v.) KA (0.1 μg/5 μl) injectio was examined in the hippocampus. Control group (CON) was injected with i.c.v. PBS. The mice number of each group was 3. Experiments were conducted independently three times, giving a total of nine per group in the final statistical analysis. ∗∗p<0.01(CON vs other groups), ∗∗∗p<0.001 (CON vs other groups).

• 

  Fig. 3. The effect of visnagin on proinflammatory cytokines increased by KA in the hippocampus. Alteration of –1β, TNF-α (B), IL-6 (C), and COX-2 (D) at 1 (A), 3 hrs (B, D), or 6 hrs (C) after intracerebroventricular (i.c.v.) KA injection was examined in the hippocampus. The increased IL-1β, TNF-α (B), IL-6 (C), and COX-2 (D) by KA was decreased by visnagin treated prior to 20 min. The mice number of each group was 3. Experiments were conducted independently three times, giving a total of nine per group in the final statistical analysis. ∗p<0.05, ∗∗p<0.01, ∗∗∗p<0.001 (CON vs KA), ++p<0.01 (Vis vs Vis + KA). CON, vehicle (i.p.) + PBS (i.c.v.); KA, vehicle (i.p.) + KA (i.c.v.); Vis, Visnagin (i.p.) + PBS (i.c.v.).

• 

  Fig. 4. The effect of visnagin on the GFAP, OX-42 expression induced by KA in hippocampal CA3 region. OX-42 and GFAP immunoreactivities (A) in the hippocampus were examined at 24 hr after i.c.v. injection of KA (0.1 μg/5 μl). Animals (N=5 per each group) were pretreated orally with either PBS or visnagin (100 mg/kg) 20 min prior to KA (0.1 μg/5 μl). To quantify, we counted GFAP or OX-42 IR in each section (B). CON (a): vehicle (i.p.) + PBS (i.c.v.), KA (b): vehicle (i.p.) + KA (i.c.v.), Vis (c): Visnagin (i.p.) + PBS (i.c.v.), Vis + KA (d): Visnagin (i.p.) + KA (i.c.v.). Antibody against OX-42 and GFAP was used at 1: 10,000 dilution for immunostaining. ∗p<0.05 (KA vs Vis + KA), ∗∗p<0.01 (KA vs Vis + KA).

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