Korean J Nutr.  2010 Aug;43(4):367-373. 10.4163/kjn.2010.43.4.367.

Anti-Oxidative and Anti-Inflammatory Effect of Water Extract from Perillae semen in RBL-2H3 Cells

• Kim JM
• Kim DJ
• Kim TH
• Kim HS
• Choe M

• KMID: 2268025
• DOI: http://doi.org/10.4163/kjn.2010.43.4.367

Abstract

The present study was to investigate anti-oxidative and anti-inflammatory activity of Perillae semen in RBL-2H3 basophilic leukemia cells. Inhibitory effect of Perillae semen onto free radical generation was determined by measuring DPPH and hydroxyl radical scavenging activities in vitro. Anti-inflammatory actions of Perillae semen extracts (100, 250, 500 microgram/mL) were assessed by testing their effects on the degranulation of mast cells. For this, beta-hexosaminidase released from RBL-2H3 cells was used and proinflammatory cytokines were measured by an ELISA kit. Our results indicated that Perillae semen water extracts effectively inhibited free radical generation. At the concentration of 500 microgram/mL of water extract, the degranulation of RBL-2H3 cells were inhibited by 42.1%. The IgE-antigen complex increased the accumulation of IL-4 and TNF-alpha secretion in RBL-2H3 cells and treatments with 250 and 500 microgram/mL of Perillae semen extracts suppressed the IgE induced secretion of IL-4 and TNF-alpha protein by 20.5, 26.9% and 14.5, 16.5% respectively. We observed that Perillae semen water extract reduced beta-hexosaminidase, IL-4, and TNF-alpha secretion in RBL-2H3 cells. These results provide that Perillae semen may be beneficial in the treatment of allergic inflammatory disease.

Keyword

perillae semen; RBL-2H3; anti-allergic; beta-hexosaminidase; IL-4; TNF-alpha

MeSH Terms

Basophils
beta-N-Acetylhexosaminidases
Cytokines
Enzyme-Linked Immunosorbent Assay
Hydroxyl Radical
Immunoglobulin E
Interleukin-4
Leukemia
Mast Cells
Perilla
Semen
Tumor Necrosis Factor-alpha
Water
Cytokines
Hydroxyl Radical
Immunoglobulin E
Interleukin-4
Tumor Necrosis Factor-alpha
Water
beta-N-Acetylhexosaminidases

Figure

• 

  Fig. 1 Effect of Perillae semen water extracts (PSWE) on DPPH radical scavenging activity. The values shown are means ± SE (n = 3). Significant differences were compared with control at *: p < 0.05 vs. control.

• 

  Fig. 2 Effect of Perillae semen water extracts (PSWE) on hydroxy radical scavenging activity. The values shown are means ± SE (n = 3). Significant differences were compared with control at *: p < 0.05 vs. control.

• 

  Fig. 3 Effect of Perillae semen water extract (PSWE) on the cell viability in RBL-2H3 cells. Results are from three experiments and are expressed as mean ± SE. Significant differences were compared with control at *: p < 0.05 vs. control, **: p < 0.01 vs. control.

• 

  Fig. 4 Effect of Perillae semen water extract (PSWE) on β-hexosaminidase release from RBL-2H3 cell. RBL-2H3 cells (2 × 105 cells) in 24-well plates were preincubated with 0.5 µg/mL anti-DNP IgE (antibody) for 12h, washed with Siraganian buffer, incubated in Siraganian buffer containing 5.6 mM CaCl2 and 0.1% BSA for 10 min, and then treated with 1 mL of PSWE for 30 min. To activate the cells and evoke an allergic reaction, the cells were stimulated for 2h with DNP-HSA (antigen; 10 µg/mL). The control group stimulated a cell with an antigen and antibody. However, without IgE-HSA group did not stimulate. β-Hexosaminidase secretion into the supernatant was then measured. Results are from three experiments and are expressed as mean ± SE. Significant differences were compared with control at *: p < 0.05 vs. control, ***: p < 0.001 vs. control.

• 

  Fig. 5 Effects of Perillae semen water extract (PSWE) on IL-4 secretion from RBL-2H3 cell. The cells were treated primed with anti-DNP IgE, treated with PSWE and stimulated with DNP-HSA as described in Fig. 4, and the supernatant was subjected to ELISA. Results are from three experiments and are expressed as mean ± SE. Significant differences were compared with control at ***: p < 0.001 vs. control.

• 

  Fig. 6 Effects of Perillae semen water extract (PSWE) on TNF-α secretion from RBL-2H3 cell. The cells were treated primed with anti-DNP IgE, treated with PSWE and stimulated with DNP-HSA as described in Fig. 4, and the supernatant was subjected to ELISA. Results are from three experiments and are expressed as mean ± SE. Significant differences were compared with control at *:p < 0.05 vs. control, **: p < 0.01 vs. control, ***: p < 0.001 vs. control.

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