Experiments were performed in triplicates, to confirm repeatability

Experiments were performed in triplicates, to confirm repeatability. lifestyle, such as improved living standards, rapid industrialization, and increased elderly population. Recent research has exploited the roles of natural materials like plants and seaweed functional foods that contribute to enhancing human health in various aspects, like improvement of liver function, sleep, and diabetes [7,8,9]. However, anthropogenic activities that contribute to the increase of harmful environmental factors like fine dust, heavy metal, microplastics, etc., caused a recent increase in abnormal cutaneous immune diseases, including atopic dermatitis (AD), asthma, immune deficiency, and immune overactivation [10]. AD is the ITI214 most commonly observed abnormal immune disease characterized by erythema, hemorrhage, edema, excoriation/erosion, scarring/dryness, and itching [11]. There are two major forms of AD, one in which the disease is usually triggered by allergens with potential immunoglobulin E (IgE) dependency, and one in which the disease appears to be IgE impartial [12]. According to the instruction of Korea Ministry of Food and Drug Safety (KMFDS), natural foods contribute to remedying or improving AD symptoms that can be used ITI214 as a functional food. Although numerous bioactive natural products of were studied, little information is usually available on the potential of for remediating hypersensitive immune responses like AD and its underlying mechanisms. The present study for the first time evaluates the effects of ethanolic extract (SHE) around the abnormal hypersensitive immune responses in human dust mite (HDM)/2,4-dinitrochlorobenzene (DNCB)-induced mice model, and its value as a functional food material. 2. Materials and Methods 2.1. Chemicals and Sample House dust mite extract ointment was purchased from Biostir Inc. (Biostir AD, Kobe, Japan). Positive control CJLP 133, a Korean FDA approved product, was purchased from CJ CheilJedang CORP (Seoul, South Korea). Dinitrochlorobenzene (DNCB) and sodium dodecyl sulfate (SDS), phosphate-buffered saline (PBS), formalin, and acetone were purchased from Sigma Chemical Co. (St, Louis, Mo, USA). IgG1 and IgG2a ELISA kits were purchased from Bethyl Laboratories (Montgomery, TX, USA). Trizol reagent was purchased from the Molecular Research Center (Montgomery, OH, USA). cDNA synthesis kit was purchased from Promega Co. (?Madison, WI, USA). Other chemicals and reagents used were the highest grades available commercially. The SHE used in this study was the same as the one used in previous studies [13,14,15]. 2.2. Mice NC/Nga female mice (8 weeks old), reared under specific pathogen-free conditions, were purchased from Orient Bio (Gwangju, Korea). The mice were housed under the conditions following; a constant temperature of 23 1.5 C, a humidity of 55 15%, and lighting followed the 12 h on/ 12 h off-cycle. Food (5L79, Orient Bio, Seongnam, Korea) and tap water were provided ad libitum for all those mice, and clean litter (BETA CHIP, Orient Bio) was used during the experiments. All mice procedures were approved by the Institutional Animal Care and Use Committee of the Chonnam National University (No.CNU IACUC-YS-2016-6). 2.3. Induction of AD and Oral Administration of SHE For the disruption of the skin barrier, the dorsal hair of the mice was shaved using an electronic shaver and hair removal cream, and applied with 150 L of Mouse monoclonal to FLT4 4% SDS, before 3 h of DNCB and HDM (AD cream, Biostir-AD, Biostir, Kobe, Japan) application. Then, DNCB and HDM were applied to the dorsal skin for the AD induction, according to the approach indicated in Physique 1. At 14 days of AD induction, the mice were randomized into six groups, as followsna?ve group (control, = 8), AD group (HDM/DNCB applied mice, = 8), SHE 10 group (HDM/DNCB and SHE 10 mg/kg co-applied mice, = 8), SHE 50 group (HDM/DNCB and SHE 50 mg/kg co-applied mice, = 8), SHE 100 groups (HDM/DNCB and SHE 100 mg/kg co-applied mice, = 8), and a P.C group (a positive control group with HDM/DNCB and CJLP 133 800 mg/kg co-applied mice, = 8). SHE was oral administrated to mice using an oral-zoned needle connected to a 1 mL syringe. Around the 35th day, the mice were dissected after the measurement of body weight. Additionally, the size and weight of spleens were ITI214 measured and used for the gene expression evaluation. CJLP 133 was used ITI214 as the positive control [16]. Open in a separate window Physique 1 Suppression of HDM/DNCB-induced AD by oral administration of SHE (10, 50, and 100 mg/kg). Schematic illustration of 6-week experimental design. 2.4. Measurement of Skin Severity Score The severity evaluation of HDM/DNCB-induced AD in.