Ayako Yamamoto

Stratum corneum lipid abnormalities in atopic dermatitis

SummaryPatients with atopic dermatitis (AD) often present with a dry skin. To clarify the relationship between dry skin and lipid abnormalities within stratum corneum, stratum corneum lipids were collected from six AD patients aged 15 to 25 years and from sex- and age-matched controls. All major stratum corneum lipid classes were separated and quantitated by high-performance thin-layer chromatography/photodensitometry. Six ceramide fractions were also isolated and quantitated by thin-layer chromatography/photodensitometry. Esterified fatty acids of both ceramide 1 (acylceramides) and wax esters were analysed by capillary gas chromatography. The relative amounts of all the stratum corneum lipid classes including squalene, cholesterol esters, wax esters, triglycerides, free fatty acids, cholesterol, ceramides, cholesterol sulphate and phospholipids did not differ statistically between AD patients and controls. However, a significant decrease in proportion of ceramide 1, which is believed to be a carrier of linoleate responsible for a water-barrier function, and increased levels of esterified C18∶1 fatty acids (oleate) of ceramide 1 were observed in AD patients. On the other hand, the fatty acid compositions as well as the proportions of C16∶1 straight-chain component in sebum wax esters of AD patients were very similar to those of controls. These results suggest that a significantly reduced amount and/or structural alterations of ceramide 1 deriving from epidermal keratinocytes may be responsible for the impaired water-barrier function of the skin in AD.

Impaired water barrier function in acne vulgaris

In acne vulgaris, abnormal follicular keratinization is important for comedo formation, yet the precise mechanisms of comedogenesis are not known. The present study examined the interrelationship between sebum secretion rate (SSR), lipid content and water barrier function (WBF) of the stratum corneum (SC) in 36 acne patients and 29 control subjects. All major SC lipid classes were separated and qantified by thin-layer chromatography/photodensitometry. WBF was evaluated by measuring transepidermal water loss (TEWL), and the hygroscopic properties and water-holding capacity of the SC. The SSR over a period of 3 h was significantly higher in patients with moderate acne than in control subjects, but no significant difference was noticed between patients with mild acne and control subjects. Significant differences between patients with both moderate and mild acne and control subjects were noted in the amount of sphingolipids (ceramides and free sphingosine), but not for any other lipid classes. Furthermore in acne patients, lower amounts of sphingolipids were observed corresponding with a diminished WBF. These results suggest that an impaired WBF caused by decreased amounts of ceramides may be responsible for comedo formation, since barrier dysfunction is accompanied by hyperkeratosis of the follicular epithelium.

Stratum corneum sphingolipids and free amino acids in experimentally-induced scaly skin

SummaryStratum corneum sphingolipids are of particular importance in maintaining the water permeability barrier of mammalian epidermis. Free amino acids also play an important role in water retention in the stratum corneum. To clarify the way in which these substances affect scaly skin, stratum corneum sphingolipids and free amino acids collected from artificially-induced scaly skin were analysed. Scaly skin was induced by tape stripping. The total amount of sphingolipids was quantified by gas chromatography and five of sphingolipid fractions were isolated and quantified by thin-layer chromatography. Free amino acids were analysed using a high-speed amino analyser. The total amount of sphingolipid in scaly skin did not differ statistically from that in control skin. However, a significant change in the distribution of the five sphingolipid species was observed in scaly skin and the total amount of amino acids was decreased in scaly skin. These results suggest that the distribution of these five types of sphingolipid and the total amount of amino acids are responsible for scaly skin.

Sebaceous gland activity and urinary androgen levels in children

Sebum and 24-h urine samples were collected from 17 children, aged 10-17 years. Wax ester (WE) secretion rates were measured as the index for sebaceous gland activity. The fatty acid methyl esters were analyzed by capillary gas chromatography. Urinary testosterone as well as 17-ketosteroids (androsterone, dehydroepiandrosterone (DHEA), and etiocholanolone) were separated from the urine samples and analyzed by gas chromatography. WE secretion rates increased with age and there were correlated changes in the monounsaturated fatty acids of WEs. Straight chain types such as 14:1 and 16:1 tended to increase, while 18:1 tended to decrease. There was a negative correlation between iso even fatty acids and WE secretion rates. However the other chain types such as straight odd, iso odd and anteiso odd showed no statistically significant change in relation to WE secretion rates. WE secretion rates were correlated positively with testosterone levels in both sexes, with DHEA in males, and with etiocholanolone in females. The results suggest that sebaceous gland activity is stimulated by adrenocortical androgens in prepuberty, in addition to the strong effect of testosterone during puberty.

Immunohistochemical distribution of cutaneous fatty acid-binding protein in human skin

Cutaneous fatty acid-binding protein (C-FABP) has been newly discovered from rat skin. The immunohistochemical study revealed that C-FABP is strongly expressed in rat epidermis. Recently, a highly homologous protein to rat C-FABP was found in psoriatic epidermis, which is termed psoriasis-associated fatty acid-binding protein (PA-FABP). In the present study, we investigated the expression of PA-FABP in normal and pathological human skin using immunohistochemical techniques. In normal skin, PA-FABP was expressed in basal and prickle cell layers, and more strongly in the granular cell layer. The expression pattern of PA-FABP was similar to that of C-FABP in rat skin. In psoriatic skin, PA-FABP was expressed in suprabasal layers and more strongly in more differentiated keratinocytes. In squamous cell carcinoma, PA-FABP showed very strong expression in squamous nests. These results indicate that PA-FABP is a human homologue of C-FABP. Moreover, it is suggested that C-FABP has important roles in the transport and metabolism of fatty acids in epidermis and that altered lipid metabolism may affect the proliferation and/or differentiation of keratinocyte.

The activity of fatty acid synthase of epidermal keratinocytes is regulated in the lower stratum spinousum and the stratum basale by local inflammation rather than by circulating hormones

The epidermal keratinocytes produce and secrete lipids to maintain the water barrier of the epidermis. To clarify the regulation of epidermal lipid synthesis, we investigated the hormonal effect on the activity of fatty acid synthase (FAS) of the keratinocytes, and the expression of FAS in the human skin. In cultured keratinocytes, the FAS activity, assayed by measuring the oxidation of NADPH, was slightly increased by hydrocortisone or testosterone, but not influenced by thyroid hormone, estrogen, progesterone or insulin. In immunohistochemical study of normal human epidermis, FAS was expressed strongly in the stratum granulosum and moderately in the uppermost layer of the stratum spinousum (SS), suggesting that fatty acid synthesis may increase during normal epidermal differentiation. In inflammatory disorders, such as psoriasis, lichen planus, and atopic dermatitis, FAS was also expressed in the lower SS and the stratum basale (SB), resulting in strong staining in the whole layers of the epidermis. Remarkable increase of FAS expression was only observed in the lower SS and the SB. Therefore, the activity of FAS in the epidermis may be regulated in the lower SS and the SB by local inflammation rather than by circulating hormones. In other components of the skin, FAS was strongly expressed not only in adipose tissue and sebaceous glands, which are known as active sites of lipid synthesis, but also in sweat glands, suggesting that the sweat glands can synthesize abundant fatty acids de novo.

Expression of cutaneous fatty acid-binding protein and its mRNA in rat skin.

Cutaneous fatty acid-binding protein (C-FABP) has been purified from rat skin. Since there was little information about the role of C-FABP in the skin, we investigated the expression of C-FABP and its mRNA in normal rat skin using an immunohistochemical technique and in situ hybridization. In the epidermis, C-FABP mRNA was found to be expressed in basal cells and highly in prickle cells, while C-FABP itself was strongly expressed in the upper prickle and the granullar cell layers. In sebaceous glands, both C-FABP and its mRNA were expressed in both peripheral and differentiating cells, although the expression of C-FABP mRNA gradually reduced during differentiation of sebocytes. Since epidermis and sebaceous glands are active sites of fatty acid synthesis, these results suggest that C-FABP may have important roles in the transport and synthesis of fatty acids.

Topical Spironolactone Reduces Sebum Secretion Rates in Young Adults

The effects of topically applied spironolactone on the sebum secretion rates (SSR) of young adults were investigated. SSR was expressed as the ratio of wax esters/[cholesterol + cholesterol esters] (WE/[C + CE]) and the amount of sebaceous lipids (squalene, triacylglycerol and wax esters). Topical spironolactone 5% gel applied to the right cheeks of the subjects produced a significant reduction in the SSR at 12 weeks (4 weeks after termination of application), but not at 8 weeks (the end of treatment). Untreated “control” areas (the left cheeks of the subjects) showed no significant change during the study. None of the subjects experienced skin rash or signs of local irritation. This results suggests that topical spironolactone may be effective in the treatment of acne patients with high SSR

High transepidermal water loss induces fatty acid synthesis and cutaneous fatty acid-binding protein expression in rat skin

Stratum corneum lipids (SCLs) are responsible for the water barrier function (WBF) of the skin in mammals. Recent studies have indicated that epidermal lipid synthesis might be regulated by WBF of stratum corneum and that cutaneous fatty acid-binding protein (C-FABP) plays an important role in fatty acid metabolism in epidermis. To clarify the regulatory mechanism of lipid synthesis, we assessed the effects of barrier disruption induced by either acetone treatment or linoleic acid deficiency on epidermal lipid synthesis in rat. Transepidermal water loss (TEWL) was measured with an evaporimeter before and after treatment. The time course of re-establishing SCLs was examined biochemically and histochemically and the expression of C-FABP in the epidermis was immunohistochemically examined. When the rat skin was covered with a vapor-impermeable membrane after the acetone treatment removing SCLs and inducing WBF disruption, the usual recovery of SCLs was blocked and TEWL was kept high by 24 h. In the uncovered area, rapid redeposition of SCLs within 24 h was found and associated with normal compositions of epidermal lipids including sphingolipids, free fatty acids and sterol and, immunohistochemically, C-FABP was very weakly expressed in epidermis at 0.5 and 2 h, and then strongly in the whole layers at 4 h, and returned to a normal pattern by 8 h. The epidermis of the covered rat skin was kept weak in C-FABP expression by 24 h. In the linoleic acid-deficient rats, TEWL did not increase and the expression pattern of C-FABP showed no notable change until 28 weeks after initiation of the diets, indicating that C-FABP expression may not be affected by altered essential fatty acid metabolism. These results suggest that increase of TEWL itself stimulates C-FABP expression, leading to activation of fatty acid metabolism.

Fatty acid composition of sebum wax esters and urinary androgen level in normal human individuals.

To investigate the effects of androgens on the fatty acid compositions of sebum wax esters, we examined sebum and urinary samples from 36 healthy individuals, aged from 3 to 59 years. The percentages of C16:1 straight chain components in wax esters were correlated positively with the urinary testosterone levels in both sexes, and with the urinary levels of etiocholanolone and total 17-ketosteroids (17-KS) in females. These data suggest that more active sebaceous glands in lipid production excrete sebum with a higher proportion of C16:1 straight chain fatty acid, which is considered to be purely endogenous. It appears, therefore, that the proportion of C16:1 straight chain fatty acid in sebum wax esters may indicate the sebaceous gland activity in both sexes. In comparison of the amounts of various straight and terminally branched fatty acids in sebum with urinary androgen levels, the straight even fatty acids tended to change in a positive correlation with testosterone levels, in contrast to the changes of the iso even fatty acids in both sexes and to those of the iso odd fatty acids in males. The straight odd fatty acids showed a similar change to that of the straight even fatty acids in males, while in females, there was no significant correlation between the amounts of the fatty acids and testosterone levels. Anteiso fatty acids showed no notable change correlated with testosterone levels. This result suggests that the synthesis of iso or anteiso fatty acids may be controlled by complex factors and that there may be a unique source of anteiso fatty acids in human sebaceous glands.