The Effect of Bacillus-Fermented Scutellariae Radix Acupuncture Solution on Chemokine and Growth Factor Production in Mouse Macrophage Stimulated by Lipopolysaccharide

경락경혈학회지 Vol.27, No.3, pp.109 118, 2010

∼ Korean Journal of Acupuncture

Wan-Su Park

Dept. of Pathology, College of Oriental Medicine, Kyungwon University

Abstract

Objectives : The purpose of this study is to investigate the effect of Bacillus-fermented Scutellariae Radix acupuncture solution (SB) on chemokine and growth factor production in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS).

Methods : Productions of chemokine and growth factor were measured by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on xMAP

technology.

Firstly, cell culture supernatant was obtained after treatment with LPS (1 ㎍ /mL) and SB for 24 hours. Then, it was incubated with the antibody-conjugated beads for 30 minutes. Detection antibody was then added and incubated for 30 minutes. After incubated for 30 minutes, strepavidin-conjugated phycoerythrin (SAPE) was added. After another 30 minutes incubation, the level of SAPE fluorescence was analyzed in Bio-plex Suspension Array System.

Results : The results of the experiment are as follows.

1. SB significantly inhibited the LPS-induced production of vascular endothelial growth factor (VEGF), interferon-inducible protein (IP)-10, and granulocyte-colony stimulating factor (G-CSF) at the concentration of 25, 50, 100, and 200 ㎍ /mL in RAW 264.7 cells (P < 0.05).

2. SB significantly inhibited the LPS-induced production of Eotaxin at the concentration of 25, 100, and 200 ㎍ /mL in RAW 264.7 cells (P < 0.05).

3. SB significantly inhibited the LPS-induced production of MIP-1α at the concentration of 25 and 100 ㎍ /mL in RAW 264.7 cells (P < 0.05).

Conclusions : These results suggest that SB has immuno-modulatory property related with its inhibition of VEGF, IP-10, G-CSF, and Eotaxin production in macrophages.

Key words : Bacillus, Fermentation, Scutellaria baicalensis, Macrophage, Chemokine, Growth factor

- 110 -

- 112 -

Fig. 1. Effect of SB on production of VEGF in RAW 264.7 cells using Bioplex cytokine assay.

Fig. 2. Effect of SB on production of IP-10 in RAW 264.7 cells using Bioplex cytokine assay.

Fig. 3. Effect of SB on production of G-CSF in RAW 264.7 cells using Bioplex cytokine assay.

- 114 - Fig. 4. Effect of SB on production of Eotaxin in RAW 264.7 cells using Bioplex cytokine assay.

α

Fig. 5. Effect of SB on production of MIP-1 in

RAW 264.7 cells using Bioplex cytokine assay.

Fig. 6. Effect of SB on production of MCP-3 in RAW 264.7 cells using Bioplex cytokine assay.

γ

Fig. 7. Effect of SB on production of IFN-

in RAW 264.7 cells using Bioplex cytokine assay.

β

α

- 116 -

α

α

α

α

- 118 -