Epidermal Growth Factor and 1α,25-Dihydroxyvitamin D3 Suppress Lipogenesis in Hamster Sebaceous Gland Cells In Vitro

Abstract

We have previously reported the establishment of a culture system of hamster auricular sebocytes. Although their morphologic and biochemical properties are very similar to those of human sebocytes, the regulation of lipogenesis is not clear. Therefore, we investigated the effect of epidermal growth factor, all-trans retinoic acid, 1alpha,25-dihydroxyvitamin D3, and androgens such as testosterone and 5alpha-dihydrotestosterone on lipogenesis in cultured hamster sebocytes. Intracellular lipid droplets detected with Oil-Red-O staining were observed in 5 d cultures and increased in a time-dependent manner; 40.7% +/- 1.11% of 2 wk cultured cells tested lipid-positive by flow cytometric analysis. When the hamster sebocytes were cultured in the presence of epidermal growth factor, all-trans retinoic acid, or 1alpha,25-dihydroxyvitamin D3, the intracellular lipid droplets were diminished by all-trans retinoic acid and epidermal growth factor, and slightly by 1alpha,25-dihydroxyvitamin D3. The intracellular lipid droplets consisted mainly of triglycerides (71.8%) and, as minor components, cholesterol (18.0%), wax esters (3.6%), and free fatty acids (6.6%). Epidermal growth factor and all-trans retinoic acid decreased the intracellular accumulation of triglycerides (92.6% and 96.1% inhibition, respectively) and free fatty acids (54.3% and 62.6% inhibition, respectively) in the sebocytes. In addition, 1alpha,25-dihydroxyvitamin D3 decreased the triglyceride level (34.3% inhibition), but augmented the accumulation of wax esters (30% increase). There was no difference in the level of cholesterol as a result of these treatments, however. In contrast, 5alpha-dihydrotestosterone augmented the formation of intracellular lipid droplets along with an increase in the accumulation of triglycerides in hamster sebocytes. Our findings that regulation of lipogenesis by all-trans retinoic acid and androgen in hamster sebocytes is identical to regulation in humans suggest that hamster sebocytes are useful for the elucidation of sebaceous function at the cellular level. Furthermore, this is the first evidence that epidermal growth factor and 1alpha,25-dihydroxyvitamin D3 may act as suppressors in the regulation of lipogenesis in hamster sebocytes in vitro.

Efficacy and Safety of Topical Application of Epidermal Growth Factor (EGF) for Korean Acne Patient

“…표피성장인자는 지방형성을 억제하여 세포 내 중성 지방 농도를 낮춘다고 알려져 있으며, 재조합 인간 표 피성장인자를 도포할 경우, 여드름양 발진이 임상적으 로 호전되고, 여드름에 도포할 경우 피지분비량이 감 소하고, 피부 수분량이 증가한다고 알려져 있다 [5][6][7]. 이전 연구들을 통해 표피성장인자 외용제가 피지분비 조절 및 여드름 병변을 호전시킬 것으로 예상되는 바, 본 연구에서는 표피성장인자를 함유한 외용제의 효과 와 안전성을 평가하고자 하였다.…”

Suppression of Sebum Production and Accumulation by <i>β</i>-Cryptoxanthin Due to the Inhibition of the Expression of Diacylglycerol Acyltransferase-1 and Perilipin in Hamster Sebocytes

“…Since hamster sebocytes constitutively produce TGs during cultivation in vitro [32], we first examined the effect of β-CRX on constitutive TG production in cultured hamster sebocytes. As shown in Figure 2(A), β-CRX was found to decrease the level of TGs in a dose-dependent manner (70% inhibition at 10 μM).…”

“…Hamster sebocytes (2.4 × 10 4 cells/cm 2 ) [32] were plated onto 96-well multiplates, 35-mm or 100-mm diameter culture dishes (Becton Dickinson, Tokyo, Japan) and then cultured for 24 h in DMEM/F12 (Invitrogen, Carlsbad, CA) supplemented with 6% heat-denatured fetal bovine serum (Nichirei Biosciences Inc., Tokyo, Japan), 2% human serum (C-C Biotech Co., Valley Center, CA), 0.68 mM L-glutamine (Invitrogen), and recombinant human epidermal growth factor (10 nM) (Progen Biotechnik GmbH, Heidelberg, Germany) to achieve complete cell adhesion as previously described [6,18]. The hamster sebocytes were treated every two days for up to 8 days with or without β-CRX (purity ≥ 95%; Shikoku Yashima Pure Chemicals, Tokushima, Japan) (Figure 1) or 13-cisRA (Sigma Chemical, St. Louis, MO) in the presence or absence of a sebocyte-differentiation inducer, insulin (10 nM) (Sigma Chemical) [12] in DMEM/F12 supplemented with heat-denatured fetal bovine serum, human serum, and L-glutamine.…”

Effects of Epidermal Growth Factor and Linoleic Acid on Lipid Contents in Human Intestinal C2BBe1 Cells

“…The secretion of triglycerides in human colon adenocarcinoma Caco-2 cells was stimulated by EGF (11). However, EGF decreased the intracellular accumulation of triglycerides in hamster auricular sebocytes (12). The stimulation of triacyl glycerol synthesis by insulin in primary cultures of adult rat hepatocytes was also suppressed by EGF through the inhibition of malic enzyme activity (13).…”

“…It was reported that EGF regulated the secretion and formation of lipids in various cells (11)(12)(13). The secretion of triglycerides in human colon adenocarcinoma Caco-2 cells was stimulated by EGF (11).…”