Won-Hong Woo

Effects of the ethanol extract of Cichorium intybus on the immunotoxicity by ethanol in mice

Effects of the ethanol extract of Cichorium intybus (CIEE) on the immunotoxicity of ethanol (EtOH) were investigated in ICR mice. Mice were divided into four groups, and CIEE at dose of 300 mg/kg was orally administered to mice daily for 28 consecutive days, and normal mice were given vehicle. Mice treated with EtOH were given freely with 20% w/v EtOH solution. The results of this study are summarized as follows: The combination of CIEE and EtOH showed significant increases in the circulating leukocytes and the relative weights of liver, spleen and thymus, as compared with those in mice treated with EtOH alone. However, the body weight gain was not affected. Splenic plaque forming cells (PFC) and hemagglutination (HA) titers to sheep red blood cells (SRBC), and the secondary IgG antibody response to bovine serum albumin (BSA) were markedly enhanced by CIEE plus EtOH treatment as compared with the treatment of EtOH alone. In mice receiving the combination of CIEE and EtOH when compared with EtOH alone-treated mice, there were also significant increases in delayed-type hypersensitivity (DTH) reaction, phagocytic activity, natural killer (NK) cell activity and cell proliferation as well as interferony (IFN-gamma) secretion. In the case of interleukin-4 (IL-4) content, however, an insignificant induction observed by CIEE plus EtOH treatment. These findings indicate that the immunotoxicity induced by EtOH is significantly restored or prevented by CIEE treatment.

Effects of the aqueous extract of Epimedii Herba on the antibody responses in mice

Antibodies and cytokines in serum were detected in male ICR mice treated with the aqueous extract of Epimedii Herba (AEEH) at doses of 40, 120 and 360 mg/kg orally for 2 weeks. Effects of AEEH on antibody forming responses were assessed by enzyme linked immunosorbent assay (ELISA) of immunoglobulin (Ig) levels in serum collected 7 days after priming with ovalbumin (OVA) in complete Freunds adjuvant (CFA) or immediately without priming at week 2. The relative spleen weight was significantly increased by AEEH, compared with controls, especially at a dose of 120 mg/kg of it after priming with OVA and 40 mg/kg without priming, respectively. However, body weight gain was slightly decreased in AEEH-fed mice. The enhancement of total serum IgG and IgG1 levels in unprimed mice was statistically significant in mice fed 40 mg/kg AEEH. Total serum IgG2a levels and Il-4 secretion were also statistically augmented by all groups of AEEH treatment. A tendency to marked increase of total serum IgM level and IFN-gamma secretion was also observed in mice fed 40 and 120 mg/kg AEEH but not those fed 360 mg/kg AEEH. When mice were immunized with OVA, furthermore, a marked stimulation of antibody formation and cytokines secretion was observed in all groups of AEEH-fed mice compared with controls. These findings indicate that AEEH at therapeutic concentrations enhances the production of antibodies and cytokines in mice, and the enhancing effects are more marked when the mice were immunized with OVA. Thus, these results suggest that AEEH is effective on Th cell functions, and protective effects on host against immune diseases.

Induction of oral tolerance to Japanese cedar pollen

Oral tolerance is thought to play a role in preventing allergic responses and immune-mediated diseases. An improved mouse model of the oral tolerance to Japanese cedar pollen (JCP) as antigen was developed in order to detect induction of the tolerance, and the immunological characteristics of this model were also elucidated. Oral tolerance was induced by C3H/ HeN mice given an oral administration of 10mg JCP 7 days before immunization with an i.p. injection of 0.1 mg JCP in complete Freunds adjuvant (CFA). The effects of oral JCP on systemic immunity were assessed by enzyme-linked immunosorbent assay (ELISA) of immunoglobulin (Ig) levels in serum collected on day 7 or 14 after immunization. Oral tolerance to JCP was adequately induced on day 7 after immunization and was more effective in C3H/HeN mice than in BALB/c mice. The tolerance was primarily concerned with the decreased serum levels of antigen-specific IgG. In these mice, oral administration of JCP also suppressed various immune responses to the antigen including delayed-type hypersensitivity (DTH), total IgE level and anti-JCP lgG1 level. The suppression of these immune responses by the oral antigen was associated with a significant reduction in interleukin-4 (IL-4) production. These findings therefore indicate that this C3H/HeN mice model has potential use in detecting the induction of oral tolerance by JCP, and suggest that this tolerance model may be effective in the treatment and prevention of allergic responses caused by the antigen.

Effect of biphenyl dimethyl dicarboxylate on the humoral immunosuppression by ethanol

The present study was undertaken to investigate the effect of biphenyl dimethyl dicarboxylate (PMC) on the humoral immunosuppression by ethanol (EtOH) in ICR mice. PMC at a dose of 6 mg/kg was orally administered to mice daily for 28 consecutive days, and the control mice were given vehicle. Mice treated with EtOH were given freely with 20% EtOH instead of water. The results of this study are summarized as follows; a gain of body weight and the relative weights of spleen and liver were significantly increased by combination of PMC and EtOH, as compared with those in mice treated with EtOH alone. Splenic plaque forming cells (PFC) and hemagglutination (HA) titers to sheep red blood cells (SRBC), and the secondary IgG antibody response to bovine serum albumin (BSA) were decreased by the treatment of EtOH alone, then restored to normal level by PMC treatment. The elevations of serum glutamic-pyruvic transaminase (S-GPT) and total protein levels caused by EtOH were reduced to normal level by the combination of PMC and EtOH. In addition, lower serum albumin and A/G ratio were also increased to normal level. These findings indicate that PMC has a protective effect against EtOH-induced humoral immunosuppression.

EPA attenuates ultraviolet radiation-induced downregulation of aquaporin-3 in human keratinocytes

Eicosapentaenoic acid (EPA) is an omega-3 polyunsaturated fatty acid (ω-3 PUFA) that protects against photodamage and photocarcinogenesis in mammals. Aquaporin-3 (AQP3) is a water/glycerol transport protein that is found in basal layer keratinocytes. In this study, we have investigated the protective effect of EPA against ultraviolet B (UVB)-induced AQP3 downregulation in human keratinocytes. EPA treatment was found to increase AQP3 gene and protein expression in human epidermal keratinocytes (HaCaT). Using a specific inhibitor, we observed that the effect of EPA on AQP3 expression was mediated by extracellular signal-regulated kinase (ERK) activation. UVB radiation induced AQP3 downregulation in HaCaT cells, and it was found that EPA treatment attenuated UVB-induced AQP3 reduction and the associated cell death. UVB-induced downregulation of AQP3 was blocked by EPA and p38 inhibitor SB203580. Collectively, the present results show that EPA increased AQP3 expression and that this led to a reduction UVB-induced photodamage.

Antiinflammatory effect of Daesiho, a Korean traditional prescription for cerebral infarct patients

Daesiho, a prescription composed of eight herbal mixtures, has been widely used in the treatment of cerebral infarct in Oriental medicine. However, the mechanisms by which the formula affects the production of pro‐inflammatory cytokines in cerebral infarct patients remains unknown. The levels of secretory protein pro‐inflammatory cytokines, including tumor necrosis factor (TNF)‐α, interleukin (IL)‐1β and IL‐6, were significantly increased in both lipopolysaccharide (LPS) and phytohemagglutinin (PHA)‐stimulated peripheral blood mononuclear cells (PBMCs) from cerebral infarct patients and LPS‐stimulated THP‐1 differentiated macrophage‐like cells (THP‐1/M). However, pretreatment with Daesiho significantly inhibited the secretion of pro‐inflammatory cytokines, including TNF‐α, IL‐1β, and IL‐6, in stimulated PBMCs and THP‐1/M cells. In addition, Daesiho significantly suppressed mRNA expression of pro‐inflammatory cytokines. Therefore, these data indicate that Daesiho may be beneficial in the cessation of inflammatory processes of cerebral infarction through suppression of the production of pro‐inflammatory cytokines via inhibition of mRNA expression. Copyright

Scopoletin stimulates melanogenesis via cAMP/PKA pathway and partially p38 activation

Scopoletin was recently shown to stimulate melanogenesis through cAMP-response element-binding protein (CREB) phosphorylation. In this study, we investigated the molecular events of melanogenesis-induced by scopoletin. After exposure to scopoletin, the protein levels of tyrosinase and tyrosianse related protein-1 (TRP-1) were significantly increased in B16F10 cells. The mRNA levels of tyrosinase and microphthalmia-associated transcription factor (MITF) were also enhanced by scopoletin. cAMP production and phosphorylation of p38 mitogen-activated protein kinase (MAPK) were increased by scopoletin treatment. Scopoletin-mediated increase of intracellular melanin and tyrosinase expression were significantly attenuated by protein kinase A (PKA) inhibitors (H-89 and KT5720), while a protein kinase C (PKC) inhibitor (Ro-32-0432) had no effect and a p38 MAPK inhibitor (SB203580) partially blocked the scopoletin-induced intracellular melanin and tyrosinase expression. Moreover, scopoletin synergistically with cell-permeable cAMP analog (dibutyryl cAMP) significantly induced tyrosinase activity and melanin content in B16F10 cells. The silencing of p38 MAPK by small interfering RNA (siRNA) decreased the scopoletin-induced tyrosinase expression in B16F10 cells. These results suggest that scopoletin could induce melanin synthesis through the cAMP/PKA pathway and partially p38 MAPK activation in B16F10 cells.