Abstract
BackgroundThe Sophora japonica extracts contain flavonol triglycoside, isoflavonol, coumarone chromone, saponin, triterpene glucoside, phospholipids, alkaloids, amino acids, polysaccharides, and fatty acids. These components have physiological effects such as anti-infertility and anti-cancer activities. This study investigated the regulation of keratinocyte differentiation upon treatment with the S. japonica extracts in keratinocyte and the molecular cell biological mechanism involved.MethodsTo determine whether the S. japonica extracts or troxerutin, which is its main component, regulates keratinocyte differentiation, quantitative real-time polymerase chain reaction (qRT-PCR) was performed on keratinocyte differentiation markers such as keratin 1 (K1), keratin 10 (K10), involucrin, and filaggrin after treatment with the S. japonica extracts. miR-181a knockdown confirmed that keratinocyte differentiation was regulated by increased miR-181a expression upon treatment with the S. japonica extracts or troxerutin.ResultsThe expression of keratinocyte differentiation markers such as K1, K10, involucrin, and filaggrin increased upon treatment with the S. japonica extracts and troxerutin. Furthermore, miR-181a expression, which is known to increase during keratinocyte differentiation, increased upon treatment with the S. japonica extracts and troxerutin. When miR-181a was knocked down, the increased expression of keratinocyte differentiation markers upon treatment with the S. japonica extracts and troxerutin decreased again. Finally, it was confirmed that miR-181a directly regulated and reduced the expression of Notch2, which reduces keratinocyte differentiation, and that the decrease in Notch2 expression by miR-181a regulated keratinocyte differentiation.ConclusionsThese results suggest that the S. japonica extracts or troxerutin accelerates keratinocyte differentiation through miR-181a. This accelerated keratinocyte differentiation was confirmed to have resulted from the regulation of Notch2 expression by miR-181a. The results of this study provide an opportunity to confirm the molecular cell biological mechanism of S. japonica extracts or troxerutin on skin keratinization, and we expected that this study contribute to develop a moisturizing cosmetic material that can strengthen the skin barrier through regulating keratinocyte differentiation.
Highlights
The Sophora japonica extracts contain flavonol triglycoside, isoflavonol, coumarone chromone, saponin, triterpene glucoside, phospholipids, alkaloids, amino acids, polysaccharides, and fatty acids
According to the mRNA levels of keratin 1 (K1), keratin 10 (K10), involucrin, and filaggrin measured from collected cells treated with 200 μg/mL of the S. japonica extract, the increased expression of keratinocyte differentiation markers due to calcium was further increased upon treatment with the S. japonica extracts (Fig. 1)
S. japonica extracts regulated miR-181a expression in HaCaT cells To determine whether keratinocyte differentiation upon treatment with the S. japonica extracts is regulated through miR-181a, the change in the expression of miR181a associated with treatment with these extracts was measured
Summary
The Sophora japonica extracts contain flavonol triglycoside, isoflavonol, coumarone chromone, saponin, triterpene glucoside, phospholipids, alkaloids, amino acids, polysaccharides, and fatty acids. These components have physiological effects such as anti-infertility and anti-cancer activities. The skin plays a role in protecting the body from external stimuli such as microbes and ultraviolet radiation (Chuong et al, 2002) These skin defense functions are mainly functions of the epidermis composed of keratinocytes and melanocytes. Basal layer keratinocyte was differentiated to corneocytes that exist in the outer layer of the epidermis, resulting in gene expression regulating differentiation, cell cycle, and cell morphology. Calcium is well known for promoting keratinocyte differentiation, which is known to increase the density of the upper epidermis (Menon et al, 1985; Hennings et al, 1980; Yuspa et al, 1989)
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