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Abstract
In our previous studies, dietary supplements of silk protein, sericin, and fibroin, were beneficial for improving epidermal levels of ceramides, which are the major lipids for maintaining the epidermal barrier. In this study, we investigated the dietary effects of silk protein on epidermal levels of free sphingoid bases and their phosphates such as C18 sphingosine (So), C18 sphinganine (Sa), C18 sphingosine-1-phosphate (S1P), and C18 sphinganine-1-phosphate (Sa1P), which are either synthetic substrate or degradative metabolites of ceramides. Forty-five male NC/Nga mice, an animal model of atopic dermatitis (AD), were divided into three groups: group CA was an atopic control and fed a control diet, group S was fed a 1% sericin diet, and group F was fed a 1% fibroin diet. Fifteen male BALB/c mice served as group C (control group) and were fed the control diet. All mice were fed with diets and water ad libitum for 10 weeks. Sa in group CA was lower than that in group C, but So in group CA was similar to that in group C. So and Sa were higher in groups S and F than those in group CA; So level was even higher than that in group C, and Sa level was similar to that of group C. The So/Sa ratio in group CA, which is reported to increase in AD, was significantly higher than that of group C. The So/Sa ratio was lower in groups S and F than that in group CA, and decreased further in group F. However, S1P and Sa1P in groups S and F were similar to those in group CA. Taken together, we demonstrated that silk protein, sericin and fibroin dietary supplements, increased So and Sa levels, and decreased the So/Sa ratio.
Figures and Tables
Fig. 1
Levels of C18 Sphingoid bases in the epidermis of mice. C: BALB/c mice were fed with a control diet, group CA, S and F: NC/Nga mice were fed with a control diet (group CA) or diets containing 1% sericin extract (group S) or 1% fibroin extract (group F) for 10 weeks. Sphingoid bases were extracted from epidermal homogenates of each groups and analyzed using HPLC. Values are presented as Means ± SEM (n = 10). Means with different alphabetical letters are significantly different according to duncan's multiple comparison at p < 0.05.
Fig. 2
Ratio of C18So/Sa in the epidermis of mice. Group C: BALB/c mice were fed with a control diet, group CA, S and F: NC/Nga mice were fed with a control diet (group CA) or diets containing 1% sericin extract (group S) or 1% fibroin extract (group F) for 10 weeks. Values are presented as Means ± SEM (n = 10). Means with different alphabetical letters are significantly different according to duncan's multiple range test at p < 0.05.
Fig. 3
Levels of C18 Sphingoidbase-1-phosphates in the epidermis of mice. Group C: BALB/c mice were fed with a control diet, group CA. S and F: NC/Nga mice were fed with a control diet (group CA) or diets containing 1% sericin extract (group S) or 1% fibroin extract (group F) for 10 weeks. Sphingoid base-1-phosphates of epidermal homogenates were first dephosphorylated by alkaline phosphatase at 37℃ then separated sphingoid bases were extracted and analyzed using HPLC. Values are presented as means ± SEM (n = 10). Means with different alphabetical letters are significantly different according to duncan's multiple range test at p < 0.05.
Table 1
Composition of experimental diets (g/kg dry diet)
1) Group C: BALB/c mice were fed with a control diet as normal control; group CA, S and F: NC/Nga mice were fed with a control diet as atopic control (group CA) or diets containing 1% sericin extract (group S) or 1% fibroin extract (group F) 2) Casein (nitrogen ×6.25), 870 g/kg 3) mix #310025(Dytes Inc, Bethlehem, PA, USA): Niacin 3 g/kg, Calcium pantothenate 1.6 g/kg, Pyridoxine HCl 0.06 g/kg, Thiamine HCl 0.6 g/kg, Riboflavin 0.6 g/kg, Folic acid 0.2 g/kg, Biotin 0.2 g/kg, Vitamin E Acetate (500 IU/g) 15 g/kg, Vitamin B12 (0.1%) 2.5 g/kg, Vitamin A Palmitate (500,000 U/g) 0.8 g/kg, VitaminD3 (400,000 IU/g) 0.25 g/kg, Vitamin K1/DextroseMix (10 mg/g) 7.5 g/kg and Sucrose 967.23 g/kg 4) Salt mix composition: AIN-93 salt mix #210025 (Dytes Inc, Bethlehem, PA, USA): Calcium Carbonate 357 g/kg, Potassium Phosphate (monobasic) 196 g/kg, Potassium Citrate H2O 70.78 g/kg, Sodium Chloride 74 g/kg, Potassium Sulfate 46.6 g/kg, Magnesium Oxide 24 g/kg, Ferric Citrate U. S. P 6.06 g/kg, Zinc Carbonate 1.65 g/kg, Manganous Carbonate 0.63 g/kg, Cupric Carbonate 0.3 g/kg, Potassium Iodate 0.01 g/kg, Sodium Selenate 0.01025 g/kg, Ammonium Paramolybdate 4H2O 0.00795 g/kg, Sodium Metasilicate 9H2O 1.45 g/kg, Chromium Potassium Sulfate12H2O 0.275 g/kg, Lithium Chloride 0.0714 g/kg, Boricacid 0.0815 g/kg, Sodium Fluoride 0.0635 g/kg, Nickel Carbonate 0.0318 g/kg, Ammonium Vanadate 0.066 g/kg, and Finely powdered sucrose221.026 g/kg
Table 2
Weight gain, food intake and FER of groups for 10 weeks
1) Group C: BALB/c mice were fed with a control diet; group CA, S and F: NC/Nga mice were fed with a control diet (group CA) or diets containing 1% sericin extract (group S) or 1% fibroin extract (group F) for 10 weeks 2) Values are expressed as means ± SEM (n = 10). Means with different alphabetical letters in the same row are significantly different according to duncan's multiple range test at p < 0.05 3) FER; Food Efficiency Ratio = gain of body weight (g)/amount of food intake (g)
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