Dietary effect of red ginseng extracts mixed with torilis fructus and corni fructus on the epidermal levels of ceramides and ceramide related enzyme proteins in uv-induced hairless mice

Yun Ju Lee; Inn Gyung Oh; Yunhi Cho

doi:10.4163/kjn.2012.45.3.211

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Lee, Oh, and Cho: Dietary effect of red ginseng extracts mixed with torilis fructus and corni fructus on the epidermal levels of ceramides and ceramide related enzyme proteins in uv-induced hairless mice

Research Article

Korean Journal of Nutrition

Korean Journal of Nutrition 2012; 45(3): 211-217.

Published online: 28 June 2012

DOI: https://doi.org/10.4163/kjn.2012.45.3.211

Dietary effect of red ginseng extracts mixed with torilis fructus and corni fructus on the epidermal levels of ceramides and ceramide related enzyme proteins in uv-induced hairless mice

Yun Ju Lee^*, Inn Gyung Oh^*, Yunhi Cho

Department of Medical Nutrition, Graduate School of East-West Medical Science, Kyung Hee University, Yongin 446-701, Korea.

To whom correspondence should be addressed. (choyunhi@khu.ac.kr)

Equal contributor:

*These authors contributed equally to this study.

Received 14 December 2011 Revised 27 January 2012 Accepted 3 April 2012

(open-access):

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

UV-irradiation is a major factor of photo-aged skin, by which pigmentation, wrinkles and laxity are increased. In addition, the epidermal barrier is disrupted, ultimately causing dryness in photo-aged skin. As an effort to search dietary sources for improving the dryness of UV irradiated skin, the dietary effect of red ginseng based functional foods on the epidermal level of ceramides, a major lipid maintaining epidermal barrier, was determined in this study. Albino hairless mice were fed either a control diet [group UV (UV-irradiated control)] or diets with 0.5% (group M0.5) or 1% (group M1.0) of red ginseng extracts mixed with Torilis fructus and Corni fructus (66.7% red ginseng) in parallel with UV irradiation for 5 wks. A normal control group (group C) was fed a control diet without UV irradiation for 5 wks. The epidermal level of ceramides in group UV was significantly lower than that in group C, in which ceramidase, an enzyme involved in ceramide degradation, was highly expressed. In group M0.5, the epidermal level of ceramide was significantly increased to the level even higher than in group C. In addition, protein expression of serine palmitoyl transferase (SPT), a key enzyme involved in de novo ceramide synthesis, was increased in group M0.5. However the epidermal levels of ceramides as well as of ceramidase protein expression in group M1.0 did not differ from those in group UV. In conclusion, we demonstrate that dietary supplementation of red-ginseng extracts mixed with Torilis fructus and Corni fructus at a level of 0.5% level in diet increased the epidermal level of ceramides coupled with the elevated expression of SPT protein.

Keywords: red-ginseng, UV-irradiation, ceramide, SPT, ceramidase

Figures and Tables

Fig. 1

Ceramide levels in the epidermis of hairless mice fed a control diets without UV irradiation for 5 weeks (group C) and UV-irradiated hairless mice fed a control diet (group UV) or diets supplemented with 0.5% (M0.5) or 1.0% (M1.0) of red ginseng extracts mixed with Torilis fructus and Corni fructus (66.7% red ginseng) in parallel with UV irradiation for 5 weeks. The ceramide levels were determined in the epidermis of group C, UV, M0.5, and M1.0 by high-performance thin-layer chromatography and expressed as ng of ceramides/µg of protein. Date are mean ± SEM (n = 10). Values without a common letter are significant different (p < 0.05) using one-way ANOVA and Duncan's multiple range test.

Fig. 2

Altered expression of SPT and ceramidase proteins in the epidermis of hairless mice fed a control diet without UV irradiation for 5 weeks (group C) and UV-irradiated hairless mice fed a control diet (group UV) or diets supplemented with 0.5% (M0.5) or 1.0%(M1.0) of red ginseng extracts mixed with Torilis fructus and Corni fructus (66.7% red ginseng) in parallel with UV irradiation for 5 weeks. A: Representative expressions of SPT and ceramidase protein in epidermis of mice. Protein extracts (15 µg each) from groups C, UV, M0.5, and M1.0 were subjected to 10% sodium dodecylsulfate-polyacrylamide gel electrophoresis and immunoblotted with polyclonal antiserum against SPT (67 kDa) or ceramidase (45 kDa) and with actin (Santa Cruz Biotechnology Inc., Santa Cruz, CA)-specific antibodies. The region of gel shown for SPT was between the 62- and 83-kDa prestained molecular markers. The region of each gel shown for ceramidase and actin was between the 34- and 55-kDa prestained molecular markers. B: The signal intensities from multiple experiments of (A) were quantified, and the integrated areas were normalized, first to the corresponding value of actin and then to the signal observed in the normal control group (group C). Data are mean ± SEM (n = 10). Values without a common letter are significant different (p < 0.05) using one-way ANOVA and Duncan's multiple range test.

Table 1

Diet composition of experimental groups (g/kg)

Composition is in units of g/kg of diet. Group C, hairless mice fed a control diet without UV irradiation for 5 weeks; groups UV, M0.5, and M1.0, hairless mice fed a control diet (UV) or diets supplemented with 0.5% (M0.5) or 1.0% (M1.0) of red ginseng extracts mixed with Torilis fructus and Corni fructus (66.7% red ginseng) contained diet in parallel with UV irradiation for 5 wks

1) AIN-93 Vitamin Mix #310025 (Dyets Inc., Bethlehem, PA)39) 2) AIN-93G salt mix 3210025 (Dyets Inc.)39) 3) Casein (nitrogen × 6.25), 870 g/kg

Table 2

Altered mRNA levels of SPT and ceramidase in the epidermis of hairless mice

1) Data are mean ± SEM value (n = 10). Group C, hairless mice fed a control diet without UV irradiation for 5 weeks; groups UV, M0.5, and M1.0, hairless mice fed a control diet (UV) or diets supplemented with 0.5% (M0.5) or 1.0% (M1.0) of red ginseng extracts mixed with Torilis fructus and Corni fructus (66.7% red ginseng) in parallel with UV irradiation for 5 weeks 2) Date are mean ± SEM (n = 10). Values without a common letter in the same row are significantly different (p < 0.05) using one-way ANOVA and Duncan's multiple range test 3) Total RNA in the epidermis of group C, UV, M0.5, and M1.0 was isolated, and real-time PCR was performed for SPT and ceramidase mRNAs. The signal intensities from real-time PCR analysis were quantified and normalized, first to the corresponding values for the GAPDH internal controls and then to that of normal control group (group C)

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