• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.356-362
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• 2010

Ceramide is important not only for the maintenance of the barrier function of the skin but also for the water-binding capacity of the stratum corneum. Although the exact role of ceramide in the human skin is not fully understood, ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries. Compared with other microorganisms, yeast is more suitable for the production of ceramide because yeast grows fast and is non-toxic. However, production of ceramide from yeast has not been widely studied and most work in this area has been carried out using Saccharomyces cerevisiae. Regulating the genes that are involved in sphingolipid synthesis is necessary to increase ceramide production. In this study, we investigated the effect of the genes involved in the synthesis of ceramide, lcb1, lcb2, tsc10, lac1, lag1, and sur2, on ceramide production levels. The genes were cloned into pYES2 high copy number vectors. S. cerevisiae was cultivated on YPDG medium at $30^{\circ}C$. Ceramide was purified from the cell extracts by solvent extraction and the ceramide content was analyzed by HPLC using ELSD. The maximum production of ceramide (9.8 mg ceramide/g cell) was obtained when the tsc10 gene was amplified by the pYES2 vector. Real-time RT-PCR analysis showed that the increase in ceramide content was proportional to the increase in the tsc10 gene expression level, which was 4.56 times higher than that of the control strain.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.192-209
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• 1995

The purpose of this study was to evaluate drug-loaded biodegradable membranes for guided tissue regeneration(GTR). The membranes were made by coating mesh of polyglycolic acid(PGA) with polylactic acid(PLA) containing 10% flurbiprofen or tetracycline. The thickness of membrane was $150{\pm}30{\mu}m$, and the pore size of surface was about $8{\mu}m$ in diameter. The release of drugs from the membrane was measured in vitro. Cytotoxity test for the membrane was performed by gingival fibroblast cell culture, and the tissue response was observed after implant of membrane into the dorsal skin of the rat for 8 wks. Ability to guided tissue regeneration of membranes were tested by measuring new bone in the calvarial defects(5mm in diameter) of the rat for 5 weeks. The amount of flurbiprofen and tetracycline released from membrane were about 30-60% during 7 days. Minimal cytotoxity was observed in the membrane except 20% drug containing membrane. In histologic finding of rat dorsal skin, many inflammatory cells were observed around e-PTFE, polyglactin 910 and PLAPGA membrane after 1 or 2 weeks. PLA-PGA membrane was perforated by connective tissue after 4 or 6 weeks, and divided as a segment at 8 weeks. In bone regeneration guiding potential test, tetracycline loaded membrane was most effective (p

• Journal of Physiology & Pathology in Korean Medicine
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• v.32 no.2
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• pp.106-112
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• 2018

The aim of this research was to determine the diverse effects of Sappan Lignum extract and brazilin on human keratinocyte HaCaT cells. We confirmed the antioxidant effect of Sappan Lignum extract and brazilin was analyzed by using an ABTS assay, confirming the efficacy of water extraction method. Also, we examined effect of Sappan Lignum extract and brazilin on the cell viability, using the MTS assay in HaCaT cells. mRNA expression levels of tight junction-related genes associated with skin barrier in HaCaT cells were analyzed using quantitative real-time PCR analysis. Sappan Lignum extract increased the cellular activity of HaCaT cells and the expression of the tight junction-related genes claudin 3, claudin 6, and ZO-2. Brazilin displayed the same effects as that of the extract on HaCaT cells activity and tight junction-related genes expression. Furthermore, dispase assay demonstrated altered cell-cell adhesion strength of Sappan Lignum extract or brazilin treated HaCaT cells. Sappan Lignum extract or brazilin might be an useful ingredient in skin-mosturizinng and anti-wrinkle cosmetics, given its effects of altering mRNA expression of tight junction-related genes and enhancing cell-cell adhesion strength of HaCaT cells.

• Korean Journal of Pharmacognosy
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• v.46 no.2
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• pp.116-122
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• 2015

Keratinocytes are first barrier against outer challenges on skin. However, it is still largely unknown about effective protectors against ultraviolet B (UVB), and oxidative stress in human keratinocyte, HaCaT cells. Inducible heme oxygenase (HO)-1 acts against oxidants that are thought to play a role in the pathogenesis of skin disorders. Therefore, the purpose of this study was to evaluate the effect of Portulaca oleracea 70% EtOH extracts against hydrogen peroxide (H₂O₂)-induced oxidative stress in human keratinocytes, HaCaT cells. P. oleracea 70% EtOH extracts showed the potent protective effects on H₂O₂-induced toxicity by induced the expression of HO-1 in human keratinocyte, HaCaT cells. Furthermore, P. oleracea 70 % EtOH extracts caused the nuclear accumulation of nuclear factor E2-related factor 2 (Nrf2) in human keratinocytes, HaCaT cells. In addition, we found that treatment with c-Jun N-terminal kinase (JNK) inhibitor (SP600125) reduced P. oleracea 70% EtOH extracts-induced HO-1 expression, and JNK inhibitor (SP600125) also inhibited protective effects by P. oleracea 70% EtOH extracts. Therefore, these results suggest that P. oleracea 70 % EtOH extracts increases cellular resistance to H₂O₂-induced oxidative injury in human keratinocyte, HaCaT cells, presumably through JNK pathway-Nrf2-dependent HO-1 expression.

• Annals of dermatology
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• v.30 no.6
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• pp.645-652
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• 2018

Background: Adiponectin, an adipokine secreted from adipocytes, affects energy metabolism and also shows anti-diabetic and anti-inflammatory properties. Recent studies have reported that adiponectin plays a role in regulating skin inflammation. Objective: This study aimed to investigate the effect of adiponectin on the expression of filaggrin (FLG) in normal human epidermal keratinocytes (NHEKs). Methods: NHEKs were serum-starved for 6h before being treated with adiponectin. Afterward, cell viability was assessed by MTT assay. We also treated with calcium, interleukin (IL)-4, and IL-13 to provide positive and negative comparative controls, respectively. Gene mRNA expression was quantified using real time reverse transcription polymerase chain reaction, and protein expression was evaluated using Western blot. To evaluate the relationship among mitogen-activated protein kinases (MAPKs), activator protein 1 (AP-1), and FLG, we also treated cells with inhibitors for MAPKs JNK, p38, and ERK1/2. Results: FLG and FLG-2 mRNA expression in NHEKs significantly increased after treatment with $10{\mu}g/ml$ adiponectin. Adiponectin also restored FLG and FLG-2 mRNA expression that was otherwise inhibited by treatment with IL-4 and IL-13. Adiponectin induced FLG expression via AP-1 and MAPK signaling. Conclusion: Adiponectin positively regulated the expression of FLG and could be useful as a therapeutic agent to control diseases related to disrupted skin barrier function.

• The Journal of Korean Obstetrics and Gynecology
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• v.28 no.3
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• pp.74-86
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• 2015

Objectives To identify whether Sovereign-Minister-Assistant-Courier (君臣佐使) Theory and Meridian Tropism (歸經) Theory of oral herbal medicine can be applied to trans-dermal medicine and whether pharmaceutical efficacy can be delivered to skin by Courier herb (使藥). Methods We reviewed previous studies related to Sovereign-Minister-Assistant-Courier Theory. And we examined whether prescription compositions based on Sovereign-Minister-Assistant-Courier Theory correspond with efficacy results of trans-dermal medicine experimentally identified. Results When Sovereign-Minister-Assistant-Courier Theory is applied to trans-dermal medicine, Courier herb act as delivering the effective components to the lesion penetrating skin barrier. And we identified that medicine absorbed through the digestive processes may not correspond with Courier herb applied topically. Conclusions We identified that Courier herb act as delivering the effective components to the lesion. But to strength evidences, additional studies on this action will be needed.

• Biomedical Science Letters
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• v.24 no.4
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• pp.357-364
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• 2018

Manganese (Mn) is used as main materials in various chemical processes of industry, but it suggested that Mn brings about its toxicant by fume or dust through respiratory system and skin barrier. Mn toxicant induces the loss of mental health and life quality by cerebrovascular and skin diseases. Nevertheless, it lefts much unknown on the mechanism and the effectively therapeutic methods about Mn toxicant. Therefore, this study was evaluated the cytotoxicity induced by manganese dioxide ($MnO_2$) in cultured NIH3T3 fibroblasts, and also, the correlation between $MnO_2$-induced cytotoxicity and oxidative stress was examined. While, the effect of Eclipta prostrata L. (EP) extract belong to Compositae was assessed against $MnO_2$-induced cytotoxicity in the view of antioxidative effect for searching the natural resources mitigating or preventing the $MnO_2$-induced cytotoxicity. In this study, $MnO_2$-induced cytotoxicity was revealed as mid-toxic by Borenfreud and Puerner's toxic criteria, and catalase (CAT), an antioxidant prevented $MnO_2$-induced cytotoxicity by the remarkable increase of cell viability in these cultures. While, in the protective effect of EP extract on $MnO_2$-induced cytotoxicity, EP extract effectively prevented the cytotoxicity induced by $MnO_2$ via antioxidative effects such as xanthine oxidase (XO) inhibitory ability and DPPH-radical scavenging ability. From the above results, EP extract showed the effective prevention against $MnO_2$-induced cytotoxicity correlated with oxidative stress by antioxidative effects. Conclusively, this study may be useful to research or development the alternatively therapeutic agent from natural resources like EP extract for the treatment of diseases resulted in oxidative stress.

• Journal of Ginseng Research
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• v.45 no.3
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• pp.456-463
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• 2021

Background: Keratinocytes form a physical barrier and act as an innate immune cell in skin. Keratinocytes secrete pro-inflammatory cytokines, such as interleukin (IL)-1β, resulting from inflammasome activation when exposed to ultraviolet (UV) irradiation. Korean Red Ginseng extracts (RGE) have been well-studied as modulators of inflammasome activation in immune cells, such as macrophages. In the study, we elucidated the role of RGE on the UV-mediated inflammasome activation in keratinocytes compared with that in macrophages. Methods: Human skin keratinocyte cells (HaCaT), human epidermal keratinocytes (HEK), human monocyte-like cells (THP-1), and mouse macrophages were treated with RGE or a saponin fraction (SF) or non-saponin fraction (NS) of RGE before and after UV irradiation. The secretion levels of IL-1β, as an indicator of inflammasome activation, were analyzed. Results: The treatment of RGE or SF in macrophages after UV irradiation inhibited IL-1β secretion, but similar treatment in HaCaT cells did not. However, the treatment of RGE or SF in HaCaT cells in the presence of poly I:C, a toll-like receptor (TLR) 3 ligand, before UV exposure elicited the inhibition of the IL-1β secretion. The inhibition was caused by the disruption by RGE or SF of the TLR mediating up-regulation of the pro-IL-1β and NLRP3 genes during the priming step. Conclusion: RGE and its saponins inhibit IL-1β secretion in response to UV exposure in both keratinocytes and macrophages. In particular, RGE treatment interrupted only the priming step in keratinocytes, although it did attenuate both the priming and activation steps in macrophages.

• Korean Journal of Veterinary Research
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• v.61 no.4
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• pp.36.1-36.8
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• 2021

The objective of this study was to evaluate the beneficial effects of the short-term application of insect-based diet in canine allergic dermatitis. Total 19 atopic dogs with concurrent cutaneous adverse food reactions were enrolled and classified into 3 groups. The treatment group (n = 7) was fed insect-based diet, the positive control group (n = 6) was fed salmon-based diet, and the negative control group (n = 6) was fed commercial or homemade diet for 12 weeks. The degree of skin lesions was evaluated based on canine atopic dermatitis extent and severity index (CADESI-4). Additionally, transepidermal water loss (TEWL) and pruritus visual analog scale were evaluated. All indices were evaluated every 4 weeks after the initial administration of hypoallergenic diets. In the treatment group, significant decrease in the CADESI-4 score was observed at 8 weeks compared to the baseline score (p = 0.031). There were significant differences in the CADESI-4 score between the groups at 8 weeks (p = 0.008), 12 weeks (p = 0.012), and TEWL at 12 weeks (p = 0.022). This preliminary result demonstrates the potential hypoallergenicity of an insect-based diet through features that diminish cutaneous lesions and skin barrier dysfunction.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.601-610
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• 2003

Biotin is a water-soluble vitamin used as a skin conditioning agent and promotes the formation of intercellular lipid layers through increased lipid synthesis, which improves the skin's natural barrier function. The anti-inflammatory effects of biotin have been investigated using in vitro assay models, such as MTT assay, measurements of concentrations of nitric oxide(NO), prostaglandin E2(PGE$_2$), and inhibition rate of 5-lipoxygenase(5-LOX). In comparison with biotin, other plant extracts were tested at the same time which were kudzu vine extract, sage extract, paeonia extract, and dipotassium glycyrrhetinate. Nitric oxide is a signal molecule with functions such as neurotransmission, local vascular relaxation, and anti-inflammation in many physiological and pathological processes. NO can cause apoptosis and necrosis of target cells such as keratinocytes and is generated from L-arginine by nitric oxide synthase (NOS). Prostanoids, including prostaglandins and thromboxanes, are generated by the phospholipase $A_2$/cyclooxygenase(COX) pathway, and leukotrienes are generated by the 5-lipoxygenase pathway from arachidonic acid. Prostaglandin E2 recently have been shown to be beneficial in the resolution of tissue injury and inflammation, also has been implicated as an immunosuppressive agent and plasma levels of PGE$_2$ are elevated in patients sustaining thermal injury. Lipoxygenase metabolites from arachidonic acid have been implicated in inflammation, anti-inflammatory activity of the raw materials was evaluated in vitro by the offered inhibition of lipoxygenase.