• Toxicological Research
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• 제35권2호
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• pp.131-136
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• 2019

Ethylhexyl dimethyl para-aminobenzoic acid (PABA) is an oily yellow liquid derivative of water-soluble PABA commonly used in sunscreen. Ethylhexyl dimethyl PABA is widely used as an ingredient in many cosmetics at an average concentration of 1.25% (0.5-2.0%) in Korea. Previous studies, including those involving animals, have demonstrated that ethylhexyl dimethyl PABA is toxic to the following four organs: testis, epididymis, spleen, and liver. In addition, experiments using human keratinocytes found that ethylhexyl dimethyl PABA inhibits cell growth and DNA synthesis at low concentrations, and halted the cell cycle of MM96L cells (human melanoma cell line) at the G1 phase. Despite limited clinical data in humans, many studies have confirmed increased mutagenicity of ethylhexyl dimethyl PABA following exposure to sunlight, which suggests that this molecule is likely to contribute to onset of sun-induced cancer despite protecting the skin through absorption of UVB. For risk assessment, the no observed adverse effect level (NOAEL) chosen was 100 mg/kg bw/day in a 4 weeks oral toxicity study. Systemic exposure dosage (SED) was 0.588 mg/kg bw/day for maximum use of ethylhexyl dimethyl PABA in cosmetics. Based on the risk assessment and exposure scenarios conducted in this study, the margin of safety (MOS) was calculated to be 180.18 for a sunscreen containing 8% ethylhexyl dimethyl PABA, which is the maximum level allowed by the relevant domestic authorities.

• 대한본초학회지
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• 제34권6호
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• pp.109-115
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• 2019

Objective : Herbal medicinal mixture (JMB) are consisted of Caryophylli Flos, Aucklandiae Radix, and Angelicae Dahuricae Radix. Each of these herbal medicines has studied on anti-aging effect in vitro. So this study was conducted to investigate efficacy and potency of JMB extract on dermal anti-aging and whitening. Methods : The JMB was extracted at room temperature by 80% ethanol. Collagenase and elastase inhibition activity in JMB ethanol extract were determined at 10, 50, 100, 500, 1000 mg/ml concentrations by colorimetric method. The toxic range of JMB ethanol extract was evaluated using MTT assay. Also, The inhibitory effect of JMB ethanol extract on tyrosinase activity and melanin contents in mouse melanoma cell line (B16F10 cell) was identified at 50, 100, 200 ㎍/㎖ levels by spectrometric assay. In each analysis, EGCG (epigallocatechin gallate) and Kojic acid were used as positive controls, respectively. Results : The elastase and collagenase inhibitory activity of JMB ethanol extract increased dose dependently. Also, The MTT assay showed that JMB, up to 400 ㎍/㎖ concentration, exhibited no toxic effect to the B16F10 cell. And following the JMB ethanol extract treat, cellular melanin contents and tyrosinase activity were dose-dependently decreased compared to those of control. Conclusion : These results suggest that JMB ethanol extract has effects to inhibitory activity on dermal wrinkle enzyme and melanogenesis. Therefore, JMB has applicable benefits for development of materials or products to have whitening and anti-aging functions on skin.

• 한국미생물·생명공학회지
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• 제30권1호
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• pp.51-56
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• 2002

국내 토양에서 분리한 식물성 병원균인 Alternaria brassicicola SW-3리 항암활성 물질 생산능을 조사하고, 활성물질을 분리 정제하여 구조를 확인하였다. A. brassicicola SW-3는potato dextrose broth를 이용하여 15$^{\circ}C$에서 2주간 진탕 배양한 다음, MTT assay를 실시하여 항암활성을 확인하였으며, 배양여액 중의 항암물질은 ethyl acetate로 추출하고, silica gel column chromatography로 정제하여 무색의 oily product를 얻었다(수율 22mg/m1). 분리된 물질은 물이나 hexane에는 녹지 않고, chloroform, ethyl acetate, ethanol 등에는 잘 녹는 특징을 보였으며, , $IR^{1}$H-NMR, $^{13}$C-NMR 등을 통해 구조를 분석한 결과, 최근에 일본에서 분리되어 항암효과가 있는 것으로 알려진 depudecin과 동일한 물질로 추정되었다. 본 실험에서 분리된 depudecin은 인체간암세포와 mouse 피부암세포에 대한 세포독성을 나타내었으며, 각각의$ IC_50$ 는 $57\mu$g/ml, $69\mu$g/ml로 나타났다. Alternaria brassicicola SW-3에 의해 생산된 물질이 기지의 물질이지만, depudecin은 아직 작용 기작이나 적용범위 등이 밝혀지지 않은 초기 연구 단계에 있는 물질로서, 새로운 항암제로서의 가능성이 매우 높아 이의 유도체를 합성하거나, 다른 항암제와의 혼용에 의해 부작용이 적은 강력한 항암제를 개발하기 위한 선도물질로 활용될 수 있을 것이다.

• 동의생리병리학회지
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• 제22권2호
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• pp.371-376
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• 2008

In this study, we have investigated the hypo-pigmentary mechanism of methanol extract of Forsythiae Fructus in human melanocyte cell line, HM3KO. Treatment of HM3KO cells with Forsythiae Fructus extract markedly inhibited melanin biosynthesis in a dose-dependent manner. Decreased melanin contents occurred through the decrease of tyrosinase protein and activity. The mRNA levels of tyrosinase and tyrosinase-related protein 1 (TRP-1) were also reduced by Forsythiae Fructus extract. Moreover, the level of intracellular cyclic AMP (cAMP) was significantly decreased by treatment of Forsythiae Fructus extract. These results suggest that Forsythiae Fructus reduces melanin synthesis by down regulation of tyrosinase mRNA transcription, and this is closely related to the cAMP-dependent pathway.

• Journal of Microbiology and Biotechnology
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• 제25권4호
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• pp.448-451
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• 2015

In a previous study, we isolated octaphlorethol A (OPA) from Ishige foliacea and evaluated its anti-melanogenesis activity in a murine melanoma cell line. However, the whitening effect and toxicity of OPA have not yet been examined in vivo. Therefore, in this study, we investigated the inhibitory effect of OPA on melanin synthesis and tyrosinase activity in an in vivo zebrafish model. More than 90% of subject embryos survived upon exposure to OPA concentrations below $25{\mu}M$, which was not significantly different from the finding in the control group. OPA markedly inhibited melanin synthesis and tyrosinase activity in a concentration-dependent manner.

• Asian Pacific Journal of Cancer Prevention
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• 제16권14호
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• pp.6073-6080
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• 2015

Background: Melanoma differentiation-associated gene-7 (MDA-7)/interleukin-24 (IL-24), a unique tumor suppressor gene, has killing activity in a broad spectrum of cancer cells. Herein, plasmids producing mda-7 proteins fused to different RGD peptides (full RGD4C and shortened RGD, tRGD) were evaluated for apoptosis induction with a hepatocellular carcinoma cell line, Hep-G2. The study aim was to improve the apoptosis potency of mda-7 by tethering to RGD peptides. Materials and Methods: Three plasmids including mda-7, mda-7-RGD and mda-7-tRGD genes beside a control vector were transfected into Hep-G2 cells. After 72 hours incubation, cell viability was evaluated by MTT assay. In addition, the rate of apoptosis was analyzed by flow cytometry using PI/annexin staining. To detect early events in apoptosis, 18 hours after transfection, expression of the BAX gene was quantified by real time PCR. Modeling of proteins was also performed to extrapolate possible consequences of RGD modification on their structures and subsequent attachment to receptors. Results and Conclusions: In MTT assays, while all mda-7 forms showed measurable inhibition of proliferation, unmodified mda-7 protein exhibited most significant effect compared to control plasmid (P<0.001). Again, flow cytometry analysis showed a significant apoptosis induction by simple mda-7 gene but not for those RGD-fused mda-7 proteins. These findings were also supported by expression analysis of BAX gene (P<0.001). Protein modelling analysis revealed that tethering RGD at the end of IL-24/Mda7 disrupt attachment to cognate receptor, IL-20R1/IL-20R2. In conclusion, fusion of RGD4C and shortened RGD peptides to carboxyl terminal of mda7, not only reduce apoptosis property in vitro but also disrupt receptor attachment as demonstrated by protein modelling.

• IMMUNE NETWORK
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• 제20권5호
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• pp.39.1-39.13
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• 2020

Sjögren's syndrome (SS) is a chronic and systemic autoimmune disease characterized by lymphocytic infiltration in the exocrine glands. In SS, type I IFN has a pathogenic role, and recently, inflammasome activation has been observed in both immune and non-immune cells. However, the relationship between type I IFN and inflammasome-associated pyroptosis in SS has not been studied. We measured IL-18, caspase-1, and IFN-stimulated gene 15 (ISG15) in saliva and serum, and compared whether the expression levels of inflammasome and pyroptosis components, including absent in melanoma 2 (AIM2), NLR family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein (ASC), caspase-1, gasdermin D (GSDMD), and gasdermin E (GSDME), in minor salivary gland (MSG) are related to the expression levels of type I IFN signature genes. Expression of type I IFN signature genes was correlated with mRNA levels of caspase-1 and GSDMD in MSG. In confocal analysis, the expression of caspase-1 and GSDMD was higher in salivary gland epithelial cells (SGECs) from SS patients. In the type I IFN-treated human salivary gland epithelial cell line, the expression of caspase-1 and GSDMD was increased, and pyroptosis was accelerated in a caspase-dependent manner upon inflammasome activation. In conclusion, we demonstrate that type I IFN may contribute to inflammasome-associated pyroptosis of the SGECs of SS patients, suggesting another pathogenic role of type I IFN in SS in terms of target tissue -SGECs destruction.

• Archives of Plastic Surgery
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• 제41권6호
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• pp.654-660
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• 2014

Background Reactive oxygen species (ROS) damages cell molecules, and modifies cell signaling. The nuclear factor E2-related factor (Nrf2) is a critical transcription regulator, which protects cells against oxidative damage. Nrf2 expression is increased in a large number of cancers. However, little information has been reported regarding the expression of Nrf2 in skin cancers. Hence, we explored the expression of Nrf2 protein in skin cancers. Methods The Nrf2 protein expression in 24 specimens, including 6 malignant melanomas (MM), 6 squamous cell carcinomas (SCC), 6 basal cell carcinomas (BCC), and 6 normal skin tissues, was evaluated by western blotting. Immunohistochemical staining was performed. The expression of Kelch-like ECH-associated protein 1 (Keap1), the key regulator of Nrf2, was also analyzed by western blotting. Results Small interfering RNA transfection to the melanoma cell line G361 confirmed that an approximately 66 kDa band was the true Nrf2 band. The western blot revealed that the Nrf2 protein was definitely expressed in normal skin tissues, but the Nrf2 expression was decreased in MM, SCC, and BCC. Immunohistochemical examination showed that expression of Nrf2 was decreased in all skin cancer tissues compared to the normal skin tissues. Keap1 was not expressed in all malignant skin tumors and normal skin tissues by western blot. Conclusions ROS was increased in various types of cancers which proteins were highly expressed or underexpressed. This study demonstrated that the expression of Nrf2 protein was down-regulated in human malignant skin tumors. We suggest that decreased expression of Nrf2 is related to skin cancers.

• 생명과학회지
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• 제29권11호
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• pp.1192-1199
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• 2019

본 연구는 화장품 소재로서 우절 에탄올추출물의 가능성을 확인하기 위한 것이다. 이를 위해 우리는 연근의 마디부분을 70%에탄올로 추출하여 사용하였으며 미백효능 및 주름개선 효과에 대한 생물학적 활성 평가를 수행하였다. 시료의 미백활성을 알아보기 위해 멜라닌세포(B16F10 cells)의 MTT assay를 이용한 샘플의 독성평가와 멜라닌 생성에 영향을 주는 관련 단백질의 발현량을 확인하였다. 시료의 주름활성억제 평가를 위해 collagenase inhibition assay를 수행하였다. 또한 UVB로 유도된 섬유아세포(CCD-986sk cells)의 MTT assay를 이용한 샘플의 독성평가와 matrix metalloprotease (MMP-1) inhibition 및 procollagen synthesis를 평가하였다. 미백활성 평가를 알아보기 위한 western bolt 결과, 멜라닌 생성과 관련된 두 단백질 Tyrosinase related protein-1 (TRP-1, Tyrosinase related protein-2 TRP-2)의 합성이 농도의존적으로 감소됨을 나타내었다. 게다가 우절 에탄올추출물의 collagenase inhibiion 실험의 결과, $500{\mu}g/ml$의 농도에서 대조군인 EGCG와 효능이 비슷한 80% 이상의 효과를 나타내었다. Procollagen synthesis 결과 UVB 자극군에 의해 콜라겐 합성은 68.8%로 감소되었으며 시료의 $25{\mu}g/ml$의 농도에서 80.2%의 콜라겐 합성에 회복능을 보였다. MMP-1 inhibition 실험결과는 $25{\mu}g/ml$ 농도에서 20.2%의 저해능을 나타내었다. 실험 결과는 NRN의 미백 및 주름 억제 효과를 검증하고, 향후 안전한 천연 화장품 재료로 사용될 수 있음을 확인했다.

• Journal of Ginseng Research
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• 제33권3호
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• pp.177-182
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• 2009

Macrophages play an important role in the first line of immunologic effects against tumor cells. The effects of nonsaponin red ginseng (NSRG) components on macrophage functions like tumoricidal activity, phagocytic activity, and NO production in young (8-weeks-old) and aged (82-weeks-old) male C57BL/6 mice were assessed in vitro, respectively. The treatment of tumor cells (melanoma B16 cells) with the supernatants of NSRG-treated macrophages resulted in an increase of cytotoxicity at 300 $\mu$g/ml in the aged mice, whereas the supernatants did not have a cytotoxic effect in the young mice. It was observed that the supernatants induced the increase of tumor cell proliferation at 150 $\mu$g/ml in the young mice, suggesting that the supernatants contain growth factors rather than cytotoxic molecules. In addition, NSRG alone had a direct cytotoxic effect on the B16 tumor cells. NSRG had no effect on the NO production by the macrophages in the young mice, while it significantly increased the level of NO release in the aged mice. There was no difference in the phagocytic activities of the macrophages by NSRG in both groups of mice. These results suggest that NSRG has differential effects on the macrophage functions in young and aged mice.