Effects of Supplementary Seleno-L-methionine on Atopic Dermatitis-Like Skin Lesions in Mice.

Effects of selenium supplementation on atopic dermatitis (AD) were investigated by administering seleno-L-methionine (SeMet) using a mouse model of AD caused by repeated application of 2,4,6-trinitrochlorobenzene (TNCB). BALB/c mice were sensitized with TNCB to the abdomen on day -7; then, TNCB was applied repeatedly to each ear three times a week from days 0 to 23. SeMet was orally administered to the mice from days 0 to 23. The efficacy of SeMet on AD was assessed by measuring ear thickness, histologic evaluation, serum total immunoglobulin (Ig) E levels, and expression of interleukin (IL)-4 in the ear and superficial parotid lymph node. Ear thickness was remarkably increased by repeated application of TNCB, and SeMet significantly suppressed ear thickness in BALB/c mice. SeMet inhibited epidermal hyperplasia and dense infiltration of inflammatory cells. The number of TNCB-induced mast cells was significantly decreased by SeMet. Serum total IgE levels that increased by the repeated application of TNCB were significantly suppressed by SeMet. Repeated application of TNCB induced expression of IL-4, a T-helper (Th) 2 cytokine, in the ear and superficial parotid lymph node of BALB/c mice and its expression was significantly inhibited by SeMet. These results demonstrated that SeMet supplementation suppresses AD-like skin lesions in BALB/c mice and inhibits the expression of total IgE and IL-4.

[The Xanthine Oxidase Inhibitory Activity and Hypouricemic Effects of Crude Drugs Obtained from the Silkworm in Mice].

This study evaluated the effects of crude drugs obtained from the silkworm in mice with oxonic acid-induced hyperuricemia using xanthine oxidase inhibitory activity and plasma uric acid levels. The plasma uric acid level was analyzed using an improved HPLC with UV detection (HPLC-UV) method, which enabled high-sensitivity analysis of a microliter of plasma. Using this method, we evaluated natural products administered orally to the hypouricemic mice. The plasma uric acid level of mice administered a water-soluble extract from silkworm larvae with botrytis (used in traditional Chinese medicine to reduce wind, lower blood pressure, and change platelet coagulation) was significantly lower than in the control group 1, 2, and 3 h after treatment. In addition, water soluble extracts from a fungus (NBRC 31161) metabolite and silkworm pupae and larvae reduced the plasma uric acid levels in mice compared with the control group.

Dietary Deficiency of Calcium and/or Iron, an Age-Related Risk Factor for Renal Accumulation of Cadmium in Mice.

The major route of cadmium (Cd) intake by non-smokers is through food ingestion. Cd is a non-essential metal absorbed through one or more transporters of essential metal ions. Expression of these transporters is affected by nutritional status. To investigate the risk factors for Cd toxicity, the effects of deficiency of essential metals on hepatic and renal accumulation of Cd were studied in mice of different ages. Mice were administered a control diet or one of the essential metal-deficient diets, administered Cd by gavage for 6 weeks, and killed; then, Cd accumulation was evaluated. Iron deficiency (FeDF) or calcium deficiency (CaDF) resulted in remarkable increases in hepatic and renal Cd accumulation compared with control-diet mice and other essential metal-deficient mice. Cd accumulation in hepatic and renal tissue was increased significantly at all ages tested in FeDF and CaDF mice. Renal Cd concentrations were higher in 4-week-old mice than in 8- and 25-week-old mice. Increase in intestinal mRNA expression of calcium transporter (CaT)1, divalent metal ion transporter-1, and metallothionein (MT)1 was also higher in 4-week-old mice than in other mice. Renal accumulation of Cd showed strong correlation with intestinal mRNA expression of CaT1 and MT1. These data suggest that CaDF and FeDF at younger ages can be a risk factor for Cd toxicity.

Oxidized dietary oils enhance immediate- and/or delayed-type allergic reactions in BALB/c mice.

BACKGROUND: The consumption of cooking oils may exacerbate some allergic diseases. In the present study, the effects of naturally oxidized olive oil on immediate- and/or delayed-type allergic reactions were investigated in BALB/c mice. METHODS: Mouse models of 3 types of allergic reactions: contact hypersensitivity (CHS), active cutaneous anaphylaxis (ACA), and DNFB-induced hypersensitivity, were orally administered naturally oxidized olive oil that was obtained by keeping the oil at room temperature for more than 3 years. The effects of ultraviolet ray (UV)-irradiated olive oil and other dietary oils as well as their possible oxidation products on CHS were also investigated. RESULTS: Naturally oxidized olive oil had a high peroxide value (POV) and exacerbated CHS, ACA, and DNFB-induced hypersensitivity in a POV-dependent manner. UV-irradiated olive oil, corn oil, sesame oil and triolein had high POVs, but almost the same acid value (AV) and thiobarbituric acid-reactive substance (TBARS) level as fresh oils. Fresh olive oil and the representative oxidation product with a high AV or TBARS level had no effect on CHS, whereas all UV-irradiated oils and naturally oxidized olive oil exacerbated it. CONCLUSIONS: Oxidized dietary oils that have high POVs exacerbated immediate- and/or delayed-type allergic reactions regardless of the different oil constituents or oxidation processes.

Effects of administering sodium selenite, methylseleninic acid, and seleno-L-methionine on glucose tolerance in a streptozotocin/nicotinamide-induced diabetic mouse model.

The effects of administering the selenocompounds, sodium selenite, methylseleninic acid (MSA), and seleno-L-methionine (SeMet) on glucose tolerance were compared in the nicotinamide (NA) and streptozotocin (STZ)-induced diabetic mouse model. ICR mice were intraperitoneally treated twice with STZ (100 mg/kg) 15 min after an injection of NA (120 mg/kg) at a 1-d interval. Non-fasting blood glucose levels were then monitored weekly while orally administering the selenocompounds at 158 µg Se/kg body weight with free access to a selenium-deficient diet for 5 weeks. The mean body weights of NA/STZ-induced diabetic mice were partly restored by the administration of selenocompounds, while SeMet led to a higher selenium content and glutathione peroxidase 1 activity in the pancreas. Non-fasting and oral glucose tolerance-tested blood glucose levels, which were elevated by NA/STZ, were significantly suppressed by the administration of SeMet. These results suggest that SeMet may improve glucose tolerance in a NA/STZ-induced mild diabetic mouse model by increasing bioavailability in the pancreas.

Effect of selenite on T-cell mitogenesis: contribution of ROS production and apoptosis signal-regulating kinase 1.

Although supplementation with the selenocompound, sodium selenite has been shown to stimulate the concanavalin A-induced T-cell mitogenic response, the mechanisms responsible remain unclear. This study was conducted to evaluate the relationships between the induction of apoptosis, formation of tumor necrosis factor (TNF)-alpha and reactive oxygen species (ROS), activation of apoptosis signal-regulating kinase (ASK) 1 and the thioredoxin (Trx) system when mitogenesis was stimulated by selenite. TNF-alpha was dose-dependently released by mouse splenocytes treated with selenite, and apoptosis was induced when TNF-alpha was added at the indicated concentrations. However, supplementation with selenite at low concentrations inhibited the accumulation of ROS with the increased expression of Trx reductase 1 and induction of apoptosis in wild-type splenocytes, and also at high concentrations in Trx-1-transgenic mouse splenocytes. The suppression of apoptosis was accompanied by a decrease in the expression of phospho-ASK1. These results suggest that the stimulation of T-cell mitogenesis by selenite may be partly attributed to the inhibited accumulation of ROS due to a reduced Trx-1/TR1 system, the inactivation of ASK1, and the suppression of apoptosis.

Low-dose ethanol aggravates allergic dermatitis in mice.

Alcohol injures dendritic cells and suppresses cellular immunity, while some evidence indicates that drinking alcohol aggravates allergic asthma. This study investigated the effect of low doses of ethanol in enhancing allergic reactions in the skin of mice. Liquid food containing alcohol was administered to conventional NC/Nga mice to induce alcoholic hepatic steatosis, and spontaneous dermatitis was evaluated. BALB/c mice were administered approximately 1 g/kg body weight of ethanol by gavage, and contact hypersensitivity (CHS) or active cutaneous anaphylaxis (ACA) was induced. Spleens were collected 24 h after the elicitation of CHS and mRNA expressions of interferon (IFN)-γ, interleukin (IL)-4, IL-6, IL-10, and IL-18 were measured by quantitative RT-PCR. Alcohol-containing diet exaggerated spontaneous dermatitis in conventional NC/Nga mice and contact hypersensitivity in BALB/c mice. Ethanol administered by gavage for 5 days enhanced contact hypersensitivity in BALB/c mice. Ethanol administration with gavage also enhanced ACA of BALB/c mice. Ethanol did not affect mRNA expression of IFN-γ and IL-4, but did enhance IL-6, IL-10, and IL-18 mRNA expression. Histological evaluation revealed an absence of hepatic steatosis in mice administered ethanol by gavage for 5 days. In ethanol-administered mice, inflamed areas presented as lesions or a local extreme accumulation of mononuclear cells in the epidermis. These findings suggest that ethanol enhances the expression of inflammatory cytokines independently from T helper (Th)1/Th2 cytokine phenotypes, causing abnormalities in the epidermis resulting in exacerbated allergic reactivity.

Effects of zinc deficiency and supplementation on leptin and leptin receptor expression in pregnant mice.

Leptin is an adipose-derived hormone that primarily regulates energy balance in response to nutrition. Human placental cells produce leptin, whereas murine placental cells produce soluble leptin receptors (Ob-R). However, the roles of these proteins during pregnancy have not been elucidated completely. As an essential metal, zinc (Zn) is central to insulin biosynthesis and energy metabolism. In the present study, the effects of Zn deficiency and supplementation on maternal plasma leptin and soluble Ob-R regulation in pregnant mice placentas were examined using enzyme-linked immunosorbent assay, reverse transcription-polymerase chain reaction, and Western blotting. Nutritional Zn deficiency significantly reduced plasma insulin concentrations and fetal and placental weights in pregnant mice. Plasma leptin concentrations in pregnant mice also increased 20- to 40-fold compared with those in non-pregnant mice. Although dietary Zn deficiency and supplementation did not affect plasma leptin concentrations in non-pregnant mice, Zn-deficient pregnant mice had significantly reduced plasma leptin concentrations and adipose leptin mRNA expression. In contrast, Zn-supplemented pregnant mice had increased plasma leptin concentrations without increased adipose leptin mRNA expression. Placental soluble Ob-R mRNA expression also decreased in Zn-deficient mice and tended to increase in Zn-supplemented mice. These results indicate that Zn influences plasma leptin concentrations by modulating mRNA expression of soluble Ob-R in the placenta, and leptin in visceral fat during pregnancy. These data suggest that both adipose and placenta-derived leptin system are involved in the regulation of energy metabolism during fetal growth.

Supplementary seleno-L-methionine suppresses active cutaneous anaphylaxis reaction.

To clarify the relationship between selenium supplementation and type I allergic reaction, we investigated the effect of seleno-L-methionine (SeMet) supplementation on the active cutaneous anaphylaxis (ACA) reaction and cytokine production in splenocytes. Female BALB/c mice were sensitized by intraperitoneal injection of ovalbumin (OVA), and SeMet was administered orally for 2 weeks followed by a challenge with OVA to induce an ACA reaction. SeMet supplementation suppressed the ACA reaction in a dose-dependent manner. Plasma OVA-specific immunoglobulin E (IgE) level was strongly inhibited in SeMet-supplemented mice compared with control mice. The mRNA expression levels of the T helper 2 (Th2) cytokines interleukin (IL)-4 and IL-13 in the spleen of SeMet-supplemented mice were lower than those in control mice. The mRNA expression level of a Th1 cytokine, interferon (IFN)-γ, in the spleen of SeMet-supplemented mice was higher than that in control mice. Splenocytes restimulated with OVA in vitro from SeMet-supplemented mice produced lower amounts of IL-4 and IL-13 than those of control mice and higher amounts of IFN-γ than those from the control mice. These results suggest that oral SeMet supplementation suppresses OVA-induced ACA reaction by lowered Th2 cytokine production and augmenting Th1 cytokine production.

Methylseleninic acid (MSA) inhibits 17ß-estradiol-induced cell growth in breast cancer T47D cells via enhancement of the antioxidative thioredoxin/ thioredoxin reductase system.

The purpose of this study was to clarify the cell growth inhibitory mechanism of human breast cancer cells caused by selenium (Se) compounds. In the presence of 17ß-estradiol (E(2)) at physiological concentrations, growth of estrogen receptor α (ERα)-positive T47D cells was markedly inhibited by 1 × 10(-6) mol/L methylseleninic acid (MSA) with no Se related toxicity.Under conditions where cell growth was inhibited, MSA decreased ERα mRNA levels and subsequent protein levels; further decreasing expression of estrogen-responsive finger protein (Efp) which is a target gene product of ERα and promotes G2/M progression of the cell cycle. Therefore, the decline in Efp expression is presumed to be involved in G2 arrest. Coincidentally, the antioxidative thioredoxin/ thioredoxin reductase (Trx/TrxR) system in cells was enhanced by the synergistic action of E(2) and MSA. It has been reported that ROS-induced oxidative stress enhanced ERα expression. E(2) increased production of intracellular ROS in T47D cells. Meanwhile, MSA significantly decreased E(2)-induced ROS accumulation. From these results, activation of the Trx/TrxR system induced by the coexistence of MSA and E(2) suppresses oxidative stress and decreases expression of ERα, and finally induces the growth arrest of T47D cells through disruption of ERα signaling.

Difference in glucose intolerance between C57BL/6J and ICR strain mice with streptozotocin/nicotinamide-induced diabetes.

Blood glucose and plasma insulin levels between C57BL/6J and ICR strain mice with nicotinamide (NA) and streptozotocin (STZ)-induced diabetes were compared to establish a suitable strain of the experimental diabetic mouse model. The mice were intraperitoneally treated twice with STZ (100 mg/kg) 15 min after injection of NA (120 mg/kg) at a 1-day interval, and non-fasting blood glucose level was then weekly monitored for 5 weeks. The blood glucose level in ICR mice gradually increased and was about 2-times higher than that in C57BL/6J mice at the end of the observation. The plasma insulin level in ICR mice was comparatively low, compared with that in C57BL/6J mice. ICR mice were also markedly glucose-intolerant when oral glucose tolerance test was performed. These results indicate that ICR strain is more sensitive than C57BL/6J strain as a mouse model with NA/STZ-induced mild diabetes.

17beta-Estradiol increases the number of effector memory CD8+ lymphocytes in mice with contact hypersensitivity and among cultured splenocytes.

To determine how estrogen exacerbates allergies, the effects of 17beta-estradiol (E2) on lymphocyte proliferation were investigated. BALB/c mice were ovariectomized, administered 3.2 microg E2, and sensitized with 50 microL 4-ethoxymethylene-2-phenyl-2-oxazolin-5-one. After 7 days, their spleens were excised and flow cytometrically analyzed. The CD8(+)CD45RA(-)CCR7(-) cell-to-CD8(+) cell ratio in the spleen was greater in the E2-administered mice than in the controls. Splenocytes were cultured under concanavalin A stimulation, with or without E2. After 4 days, the above ratio was greater in the case of E2-treated splenocytes. E2 increases the number of effector memory CD8(+) lymphocytes during sensitization in contact hypersensitivity.

17beta-Estradiol enhances expression of inflammatory cytokines and inducible nitric oxide synthase in mouse contact hypersensitivity.

The effects of 17beta-estradiol (E2) on the expression of cytokines, inducible nitric oxide synthase (iNOS), and cyclooxygenase (COX)-2 in mouse contact hypersensitivity (CHS) were examined. Three week old female mice were ovariectomized, administered 3.2 microg of E2 subcutaneously, the mice sensitized by application of 4-ethoxymethylene-2-phenyl-2-oxazolin-5-one (OXA) on their backs, and CHS induced by applying OXA to the auricles. E2 significantly increased mRNA expression of interferon (IFN)-gamma and interleukin (IL)-10 in the auricle at 6 and 24 h after allergy elicitation in the ear, respectively, when compared to untreated controls. Although there was no effect of E2 on the expression of IL-4 and COX-2 at any time, the expression of iNOS mRNA was increased by E2 treatment at 48 h after elicitation. E2 also enhanced the expression of tumor necrosis factor (TNF)-alpha and IL-1beta. Histological evaluation revealed that E2 promoted edema of the auricle dermis. The hyperplasia of the epidermis was suppressed by E2 and the cell infiltration observed after elicitation was not altered by E2. These results suggest that E2 enhances the expression of IFN-gamma, TNF-alpha, and IL-1beta to augment the edema of auricle dermis in mouse CHS.

17beta-estradiol enhances contact hypersensitivity in a hair cycle-dependent manner and retards thymic atrophy with age.

The effect of 17beta-estradiol (E2) on murine contact hypersensitivity (CHS), thymic atrophy, and hair-cycle change related to growth was investigated. Female mice were ovariectomized. E2 (3.2 microg) was injected subcutaneously along with the sensitizer 4-ethoxymethylene-2-phenyl-2-oxazolin-5-one (OXA), and a hypersensitive reaction was elicited with OXA on the ear in mice of various ages. E2 enhanced allergy only in 3- and 7-week-old mice, just prior to hair regrowth. Age-related thymus atrophy was repressed in the E2-treated mice compared with the control mice. E2 alone did not cause thymus involution, but it did inhibit thymus involution and regeneration after CHS.

17beta-estradiol enhances contact hypersensitivity and IFN-gamma expression in inflamed skin of BALB/c mice.

The effects of 17beta-estradiol (E(2)) on mouse contact hypersensitivity (CHS) elicited at the ears by 4-ethoxymethylene-2-phenyl-2-oxazolin-5-one (OXA) were examined. Male and female BALB/c mice were sham-treated or gonadectomized, and then subcutaneously injected with E(2) twice a week for 4 weeks. The mice were sensitized by OXA application to their back and CHS was elicited at the ears. E(2) enhanced the ear swelling of all groups at 6h after the elicitation. E(2) had no effect on the mitogenesis of splenic lymphocytes or nitric oxide synthesis by peritoneal macrophages. E(2) increased the number of thymic cells in female mice, but not male mice, and had no effect on the splenic cells of either female or male mice. Evaluation of the cytokine expressions in the inflamed skin revealed that E(2) enhanced the expression of interferon-gamma, but had no effect on the expression of interleukin-4. These results suggest that E(2) affects the thymus and enhances the production of interferon-gamma in skin to augment the skin swelling in CHS elicited by OXA.