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Abstract
Ginsan, a botanic polysaccharide extracted from Panax ginseng, has recently been reported to modulate mucosal immune response. In this study, we investigated the protective effect of Ginsan against fatal Vibrio vulnificus mucosal infection. A lethal dose of V. vulnificus (1.0 × 106 CFU/mouse) was nasally inoculated to mice. The bacterial count in the nasal associated lymphoid tissue (NALT) of the mouse was significantly reduced in the Ginsan-treated group. The Ginsan-treated group showed improved survival compared to the control group (100% vs 18%). To elucidate the effect of Ginsan on modulating host immune response, cytokine mRNA expressions involved in mediating inflammation were determined by semiquantitative RT-PCR in the NALTs of the infected mice. Most of the cytokine mRNAs were similarly expressed as the control group. However, COX-1 mRNA expression level was higher in Ginsan-treated group compared to the control group. The protective effect of Ginsan was antagonized by treating with a specific COX-1 inhibitor, SC-560. Thus, these data suggest that the protective effect of Ginsan against V. vulnificus infection is partly mediated by modulating COX-1 expression.
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Figure 1.
Effect of Ginsan on V. vulnificus intranasal infection. (A) BALB/c mice were pretreated with Ginsan (100 mg/kg) for two days, control mice with PBS. On the 3rd day, mice were nasally inoculated with 1 × 106 V. vulnificus in 10 μl PBS. Mice were killed 4 hrs after infection and the number of V. vulnificus in the NALT was determined by plating on HI agars. (∗p <0.05) (B) Mouse survival following intranasal V. vulnificus infection. Mice were observed for 7 days and live mice were counted (each group consists of 6 mice).
Figure 2.
Semiquantitative RT-PCR for mRNA expression of inflammatory cytokine genes. (A) BALB/c mice were treated with Ginsan (100 mg/kg) for two days, control mice with PBS. On the 3rd day, mice were nasally inoculated with 1 × 106 V. Vulnificus in 10 μl PBS. Mice were killed 2 hrs after infection and a semiquantitative RT-PCR for various inflammatory cytokine mRNA expressions was performed. (B) A semiquantitative RT-PCR for COX-1 mRNA from the NALT at 1, 2, and 4 hrs following V. vulnificus intranasal infection.
Figure 3.
Effect of Ginsan and/or SC-560 pretreatment on V. vulnificus intranasal infection. BALB/c mice were treated with Ginsan (100 mg/kg) for two days, control mice with PBS. On the 3rd day, 1 hr before bacterial challenge, SC-560 (2.5 mg/kg), a specific COX-1 inhibitor, was orally administered. And then mice were nasally inoculated with 1 × 106 V. Vulnificus in 10 μl PBS and were sacrificed 4 hrs after infection to enumerate the number of the bacteria in the NALT (∗p < 0.05: control vs Ginsan).
Table 1.
Primers used for semiquantitative RT-PCR
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