Ceramides play major roles in maintaining the epidermal barrier. It has been sus-pected that the depletion of ceramides, associated with disrupted barrier function in the epidermis, leads to the clinical manifestation of dryness and inflammation seen in patients with psoriasis. The aim of the present study was to determine the relation-ship between the level of ceramide synthesis in the epidermis and the clinical severity in patients with psoriasis. Samples from lesional and unlesional epidermis obtained from psoriasis patients were incubated with [14C]serine, an initiator of ceramide syn-thesis. otal ceramide was fractionated using high performance thin layer chromato-graphy, and the radioactivity was measured. The clinical severity of psoriasis was graded according to the psoriasis area and severity index scoring system. The level of ceramide synthesis in the lesional epidermis of patients was significantly lower than that in the unlesional epidermis and bore a negative correlation with the clinical severity of psoriasis. The present results suggest that the decreased level of ceramide synthesis in the epidermis contributes to the clinical severity of psoriasis.
Adolescent ; Adult ; Biological Markers ; Ceramides/*metabolism ; Fatty Acids/*metabolism ; Female ; Humans ; Korea/epidemiology ; Male ; Psoriasis/classification/epidemiology/*metabolism/*pathology ; Severity of Illness Index ; Skin/*metabolism/*pathology ; Statistics

Phytoestrogens, especially Yak-kong or soybean-derived isoflavones have been traditionally used as a supplement of estrogen for preventing postmenopausal osteoporosis in oriental folk medicine. In a previous study, we demonstrated that as Yak-kong and soybean increased MG-63 human osteoblastic cell proliferation, the expression of estrogen receptor alpha and beta (ERalpha: ERbeta) both were increased. However, the increased level of ERalpha is much higher than that of ERbeta. To determine whether the altered level of ERalpha expression affects Yak-kong or soybean induced MG-63 cell proliferation, we established cell lines stably expressing either ERalpha or antisense ERalpha RNAs. Increased expression of ERalpha in MG-63 cells (ERalpha-MG63) enhanced Yak-kong or soybean induced proliferation which paralleled with the enhanced expression of IGF-I. Inhibition of ERalpha expression by antisense ERalpha RNAs (As-ERalpha-MG63) caused these cells to insensitize Yakkong or soybean induced proliferation and IGF-I expression. Furthermore, the comparable effects between Yak-kong and the combined treatment of genistein and daidzein at 0.5 x 10-8M, which is a concentration of these two isoflavones similar to Yak-kong at 0.001 mg/ml, on cell proliferation and IGF-I expression in ERalpha-MG63 or As-ERalpha-MG63 cells demonstrate that ERalpha plays an important, active role in MG-63 cell proliferation induced by phytoestrogens, especially Yak-kong or soybean derived isoflavones.
Cell Line ; Cell Proliferation ; Estrogen Receptor alpha ; Estrogen Receptor beta ; Estrogens* ; Female ; Genistein ; Humans* ; Insulin-Like Growth Factor I ; Isoflavones ; Medicine, Traditional ; Osteoblasts* ; Osteoporosis, Postmenopausal ; Phytoestrogens ; RNA ; Soybeans*

Ceramides are the main lipid component maintaining the lamellae structure of stratum corneum, as well as lipid second messengers for the regulation of cellular proliferation and/or apoptosis. In our previous study, psoriatic skin lesions showed marked decreased levels of ceramides and signaling molecules, specially protein kinase C-alpha (PKC-alpha) and c-jun N-terminal kinase (JNK) in proportion to the psoriasis area and severity index (PASI) scores, which suggested that the depletion of ceramide is responsible for epidermal hyperproliferation of psoriasis via downregulation of proapoptotic signal cascade such as PKC-alpha and JNK. In this study, we investigated the protein expression of serine palmitoyltransferase (SPT) and ceramidase, two major ceramide metabolizing enzymes, in both psoriatic epidermis and non-lesional epidermis. The expression of SPT, the ceramide generating enzyme in the de novo synthesis in psoriatic epidermis, was significantly less than that of the non-lesional epidermis, which was inversely correlated with PASI score. However, the expression of ceramidase, the degradative enzyme of ceramides, showed no significant difference between the lesional epidermis and the non-lesional epidermis of psoriatic patients. This might suggest that decreased expression of SPT protein is one of the important causative factors for decreased ceramide levels in psoriasis.
Adolescent ; Adult ; Amidohydrolases/*biosynthesis/metabolism ; Apoptosis ; Cell Proliferation ; Ceramidases ; Ceramides/chemistry ; Child ; Epidermis/metabolism ; Female ; Humans ; JNK Mitogen-Activated Protein Kinases/metabolism ; Male ; Models, Biological ; Protein Kinase C-alpha/metabolism ; Psoriasis/*blood/diagnosis ; Serine C-Palmitoyltransferase/*biosynthesis

PURPOSE: Borage oil (BO) and safflower oil (SO) are efficacious in reversing epidermal hyperproliferation, which is caused by the disruption of epidermal barrier. In this study, we compared the antiproliferative effect of dietary BO and SO. Altered metabolism of ceramide (Cer), the major lipid of epidermal barrier, was further determined by measurement of epidermal levels of individual Cer, glucosylceramide (GlcCer), and sphingomyelin (SM) species, and protein expression of Cer metabolizing enzymes. METHODS: Epidermal hyperproliferation was induced in guinea pigs by a hydrogenated coconut diet (HCO) for 8 weeks. Subsequently, animals were fed diets of either BO (group HCO + BO) or SO (group HCO + SO) for 2 weeks. As controls, animals were fed BO (group BO) or HCO (group HCO) diets for 10 weeks. RESULTS: Epidermal hyperproliferation was reversed in groups HCO + BO (67.6% of group HCO) and HCO + SO (84.5% of group HCO). Epidermal levels of Cer1/2, GlcCer-A/B, and beta-glucocerebrosidase (GCase), an enzyme of GlcCer hydrolysis for Cer generation, were higher in group HCO + BO than in group HCO, and increased to levels similar to those of group BO. In addition, epidermal levels of SM1, serine palmitoyltransferase (SPT), and acidic sphingomyelinase (aSMase), enzymes of de novo Cer synthesis and SM hydrolysis for Cer generation, but not of Cer3-7, were higher in group HCO + BO than in group HCO. Despite an increase of SPT and aSMase in group HCO + SO to levels higher than in group HCO, epidermal levels of Cer1-7, GlcCer-A/B, and GCase were similar in these two groups. Notably, acidic ceramidase, an enzyme of Cer degradation, was highly expressed in group HCO + SO. Epidermal levels of GlcCer-C/D and SM-2/3 did not differ among groups. CONCLUSION: Dietary BO was more prominent for reversing epidermal hyperproliferation by enhancing Cer metabolism with increased levels of Cer1/2, GlcCer-A/B, and SM1 species, and of GCase proteins.
Animals ; Borago* ; Carthamus tinctorius* ; Ceramidases ; Cocos ; Diet ; Epidermis* ; Glucosylceramidase ; Guinea Pigs* ; Guinea* ; Hydrogen ; Hydrolysis ; Metabolism* ; Safflower Oil* ; Serine C-Palmitoyltransferase ; Sphingomyelin Phosphodiesterase

PURPOSE: Skin pH, an indicator of skin health, is maintained by various organic factors, which include lactate, free amino acid (FAA), and free fatty acid (FFA). As skin ages or with illness, skin pH becomes less acidic, and functional food has been developed to maintain the acidic pH of skin. In this study, we determined the dietary effect of green tea extract (GTE) on skin pH of photo-aged mice, as measured by epidermal levels of lactate, FAA, and FFA. The protein expression and activity of lactate dehydrogenase (LDH), an enzyme of pyruvate reduction for lactate generation, was further determined. METHODS: Albino hairless mice were fed a control diet (group UV+) or a diet with 1% GTE (group GTE) in parallel with UV irradiation for 10 weeks. A normal control group was fed a control diet without UV irradiation for 10 weeks (group UV-). RESULTS: Skin pH was higher (less acidic) in group UV+ than in group UV-. In parallel, epidermal levels of lactate and FFA, as well as of LDH protein expression and activity, were reduced in group UV+. Dietary supplementation of GTE (group GTE) reduced skin pH to similar to the level of group UV-, and inversely increased epidermal levels of lactate, LDH protein expression and activity, but not of FFA. Although epidermal levels of FAA were similar in groups UV- and UV+, it was increased in group GTE to a level higher than in group UV-. In further analysis of major FFA, epidermal levels of palmitic acid [16:0], oleic acid [18:1(n-9)], and linoleic acid [18:2(n-6), but not of stearic acid [18:0] in group GTE were similar to or lower than those in group UV+. CONCLUSION: Dietary GTE normalized skin pH with increased levels of lactate and FAA, as well as with increased protein expression and activity of LDH in the epidermis of UVB irradiated hairless mice.
Animals ; Diet ; Dietary Supplements ; Epidermis ; Functional Food ; Hydrogen-Ion Concentration* ; L-Lactate Dehydrogenase* ; Lactic Acid* ; Linoleic Acid ; Mice ; Mice, Hairless* ; Oleic Acid ; Palmitic Acid ; Pyruvic Acid ; Skin* ; Tea*

Ceramides are the main lipids in the stratum corneum and are generated during cellular stress and apoptosis by de novo synthesis or by the action of sphingomyelinase. In addition, they are lipid second messengers produced by sphingolipid metabolism and trigger important cell responses, including protein kinase C-alpha (PKC-alpha) activation and the stimulation of signal transduction pathways with apoptosis and stress-activated protein kinases (SAPK), such as c-jun N-terminal kinase (JNK). Thus, ceramides have anti-proliferative and apoptotic effects. This study measured the changes in the levels of epidermal ceramides and ceramide-related apoptotic signaling molecules in psoriasis patients. Samples from lesional and non-lesional epidermis were obtained from psoriasis patients. Total ceramides were fractionated using thin-layer chromatography, and the levels of PKC-alpha and JNK expression were measured using Western blot analysis with specific antibodies. The ceramide level was reduced significantly, and this was associated with the downregulation of apoptotic signaling molecules, such as PKC-alpha and JNK, in the lesional epidermis of psoriasis patients. These results suggest that the decreased level of ceramides downregulates the apoptotic pathway, leading to epidermal proliferation in psoriasis.
Adult ; Apoptosis/physiology ; Blotting, Western ; Ceramides/*metabolism ; Chromatography, Thin Layer ; Epidermis/metabolism/pathology/physiopathology ; Female ; Humans ; JNK Mitogen-Activated Protein Kinases/metabolism ; Male ; Protein Kinase C-alpha/metabolism ; Psoriasis/metabolism/*pathology/physiopathology ; Severity of Illness Index ; Signal Transduction/*physiology

PURPOSE: Ultraviolet (UV) irradiation decreases epidermal hydration, which is maintained by reduction of natural moisturizing factors (NMFs). Among various NMFs, free amino acids (AA) are major constituents generated by filaggrin degradation. This experiment was conducted to determine whether or not dietary supplementation of green tea extract (GTE) in UV-irradiated mice can improve epidermal levels of hydration, filaggrin, free AAs, and peptidylarginine deiminase-3 (PAD3) expression (an enzyme involved in filaggrin degradation). METHODS: Hairless mice were fed a diet of 1% GTE for 10 weeks in parallel with UV irradiation (group UV+1%GTE). As controls, hairless mice were fed a control diet in parallel with (group UV+) or without (group UV-) UV irradiation. RESULTS: In group UV+, epidermal levels of hydration and filaggrin were lower than those in group UV-; these levels increased in group UV+1% GTE to levels similar to group UV-. Epidermal levels of PAD3 and major AAs of NMF, alanine, glycine and serine were similar in groups UV- and UV+, whereas these levels highly increased in group UV+1% GTE. CONCLUSION: Dietary GTE improves epidermal hydration by filaggrin generation and degradation into AAs.
Alanine ; Amino Acids ; Animals ; Diet ; Dietary Supplements ; Epidermis* ; Glycine ; Metabolism* ; Mice ; Mice, Hairless* ; Serine ; Tea*

The objective of this study was to evaluate whether nutrient intakes and serum biochemical indices are associated with the skin condition of humans. Anthropometric data, skin surface hydration, ceramide content, dietary intake of nutrients and serum lipids (total cholesterol, HDL-, LDL-cholesterol, TG, MDA: malondialdehyde, TAS: total antioxidant status) were obtained from 36 healthy young women. Subjects were classified into 2 groups {LM: Low Moisture Group (n = 20), HM: High Moisture Group (n = 16)} by forehead skin hydration. The results of this study were as follows: The average age, BMI, total body water, LBM, body fat%, WHR, forehead hydration, ceramide content were 20.2 yrs, 20.7, 28.8 l, 39.2 kg, 28.0%, 0.8, 44.3%, and 1.05 microgram/microgram protein, repectively. The intakes of SFA (saturated fatty acid), MUFA (monounsaturated fatty acid) in the HM group were significantly higher (p < 0.05) than in the LM group, but zinc intakes in the LM group were higher than in the HM group. Serum levels of TAS in the LM group (1.27 mmol/l) were significantly higher (p < 0.05) than that of in the HM group (1.20 mmol/l). Whereas other lipid levels were not significantly different, intakes of vitamin B6 and folate showed significant positive correlation with the forehead hydration (r = 0.447, r = 0.377). Intakes of calcium and phosphorus showed significant negative correlation with forearm ceramide content (r = -0.496, r = -0.485). Several associations between nutrient intakes and skin conditions were observed, indicating that changes in baseline nutritional status may affect skin health.
Body Water ; Calcium ; Cholesterol ; Female* ; Folic Acid ; Forearm ; Forehead ; Humans ; Malondialdehyde ; Nutritional Status ; Phosphorus ; Skin* ; Vitamin B 6 ; Zinc

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.
Animals ; Ceramidases ; Ceramides ; Cornus ; Diet ; Dietary Supplements ; Functional Food ; Mice ; Mice, Hairless ; Panax ; Pigmentation ; Proteins ; Serine ; Skin ; Transferases

Linoleic acid [LA; 18: 2 (n-6)] is the most abundant polyunsaturated fatty acid in human skin. The exclusion of LA from diet induces epidermal hyperproliferation, which is reversible by the inclusion of LA in diet, and hence, LA is heralded as an essential fatty acid (EFA). Since safflower oil (SO) has been widely recognized as the major dietary source of LA and Arctii Fructus (Arctium lappa L.) is recently reported to contain high level of LA, we compared the antiproliferative effects of SO and Arctii Fructus in this study. Epidermal hyperproliferation was induced in guinea pigs by hydrogenated coconut oil (HCO) diet for 8 wk. During following 2 wk, EFA deficient guinea pigs were fed diets of safflower oil (group HS), water extract of Arctii Fructus (group AW) or organic extract of Arctii Fructus (group AO). Normal control group was fed SO containing diet (group SO) and EFA deficient group was fed HCO containing diet (group HCO) for 10 wk. Epidermal hyperproliferation was reversed in groups AO (55.9% of group HCO) and HS(74.1% of group HCO). However, the thymidine incorporation into epidermal DNA of group HS was greater than of normal control group SO. Epidermal hyperproliferation was not reversed in group AW. The accumulations of LA into phospholipids and ceramides, and of 13-hydroxyoctadecadienoic acid (13-HODE), the potent antiproliferative metabolite of LA in the epidermis of group AO were greater than of group HS. In contrast, the de novo synthesis of ceramides, the major lipids maintaining epidermal barrier, did not differ between all of groups. Together, our data demonstrate that organic extract of Arctii Fructus is more prominent than safflower oil in reversing epidermal hyperproliferation by inducing the higher accumulations of LA and 13-HODE in the epidermis of guinea pigs.
Animals ; Ceramides ; Cocos ; Diet ; DNA ; Epidermis ; Guinea Pigs* ; Guinea* ; Humans ; Hydrogen ; Linoleic Acid* ; Phospholipids ; Safflower Oil ; Skin ; Thymidine ; Water