• The Korean Journal of Physiology and Pharmacology
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• v.25 no.1
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• pp.87-94
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• 2021

Skin photoaging occurs due to chronic exposure to solar ultraviolet radiation (UV), the main factor contributing to extrinsic skin aging. Clinical signs of photoaging include the formation of deep, coarse skin wrinkles and hyperpigmentation. Although melanogenesis and skin wrinkling occur in different skin cells and have different underlying mechanisms, their initiation involves intracellular calcium signaling via calcium ion channels. The ORAI1 channel initiates melanogenesis in melanocytes, and the TRPV1 channel initiates MMP-1 production in keratinocytes in response to UV stimulation. We aimed to develop a drug that may simultaneously inhibit ORAI1 and TRPV1 activity to help prevent photoaging. We synthesized nootkatol, a chemical derivative of valencene. TRPV1 and ORAI1 activities were measured using the whole-cell patch-clamp technique. Intracellular calcium concentration [Ca2+]i was measured using calcium-sensitive fluorescent dye (Fura-2 AM). UV-induced melanin formation and MMP-1 production were quantified in B16F10 melanoma cells and HaCaT cells, respectively. Our results indicate that nootkatol (90 μM) reduced TRPV1 current by 94% ± 2% at -60 mV and ORAI1 current by 97% ± 1% at -120 mV. Intracellular calcium signaling was significantly inhibited by nootkatol in response to ORAI1 activation in human primary melanocytes (51.6% ± 0.98% at 100 μM). Additionally, UV-induced melanin synthesis was reduced by 76.38% ± 5.90% in B16F10 melanoma cells, and UV-induced MMP-1 production was reduced by 59.33% ± 1.49% in HaCaT cells. In conclusion, nootkatol inhibits both TRPV1 and ORAI1 to prevent photoaging, and targeting ion channels may be a promising strategy for preventing photoaging.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.4
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• pp.365-370
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• 2016

Atopic dermatitis refers to a chronic, recurrent, skin condition, typically typified by itching, inflamed skin. It precedes other allergic diseases, such as asthma, food allergies, and allergic rhinitis, and is usually accompanied by various other immune disorders and secondary symptoms. In this study, we discovered that when treating TNF-${\alpha}$ and IFN-${\gamma}$-stimulated HaCaT cells with various concentrations of Picea wilsonii Mast (PwM) extracts, the cell viability was excellent. In addition, we measured the inflammatory cytokines associated with atopic dermatitis, including IL-6, IL-8, IL-13, and MCP-1. The production of IL-6, IL-13, and MCP-1 decreased in the presence of PwM extracts, whereas there was no significant difference in the production of IL-8. Further studies are necessary to develop an effective cure for atopic dermatitis and inflammation using foreign plant extracts, and PwM efficacy should be determined with an in-depth, objective verification process using protein and mechanism analysis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.27 no.1
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• pp.17-27
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• 2001

A cell-based assay system for monitoring NF-$textsc{k}$B activity was developed to determine the influence of activated NF-$textsc{k}$B in human HaCaT cells. The pNF-$textsc{k}$B-SEAP-NPT plasmid that permits expression of the secreted alkaline phosphatase (SEAP) reported gene in response to the NF-$textsc{k}$B activity and contains neomycin phosphotransferase (NPT) gene for the geneticin resistance in host cells was constructed and transfected into human keratinocyte cell line HaCaT. Human HaCaT transfectant cells secreted the SEAP enzyme into the culture medium in a time-dependent manner until 72h. NF-$textsc{k}$B activities were measured in the SEAP reporter gene assay using a fluorescent detection method. The treatment of HaCaT cell transfectants with known antioxidants [e.g., N-acetyl-L-cysteine and vitamin C] showed inhibition of NF-$textsc{k}$B activity in a time-and concentration-dependent manner. The phorbol 12-myristate 13-acetate (PMA) known as a stimulator of NF-$textsc{k}$B expression demonstrated that it increased NF-$textsc{k}$B activity in a time- and concentration-dependent manner. This assay system could be used to determine the quantitative measurement of NF-$textsc{k}$B activity in the human skin and allow the screening of anti-inflammatory agents from various synthetic chemicals and natural products for dermatological purpose. Abbrevitions used: NF-$textsc{k}$B, nuclear factor kappa B; I-$textsc{k}$B, Inhibitory kappa B; SEAP, secreted alkaline phosphatase; NPT, neomycin phosphotransferease; PCR, polymerase chain reaction: dNTP, deoxynucleoside triphosphates; DMEM, dulbecco’s modified eagle medium; FBS, fetal bovine serum; PBs, phosphate-buffered saline; MUP, 4-methylumbellifery phosphate; NAC, N-acetyl-L-cysteine; DMSO, dimethyl sulfoxide; PMA, phorbol 12-myristate 13-acetate.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.3
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• pp.82-97
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• 2007

Objective : Cortex of Ulmi pumilae(CUP) has been used to treat several diseases including boil, swelling, and scabies etc. Recently, CUP was known to have wrinkle care and whitening actions. But, It's exact mechanisms are unclear. Methods : The present study was designed to investigate effects of CUP on Human HaCaT keratinocyte and malignant melanoma cells such as SK-MEL-2 and B16F10 in terms of cell viabilities, proliferations, DPPH free radical scavenging activities, oxygen free radical productions and inhibitory action on elastase activities. Results : CUP accelerated proliferation of HaCaT keratinocytes in the lower concentration. CUP also prevented cell death of HaCaT induced by Hydrogen peroxide, which products oxygen free radicals. On the contrary, CUP did not affect proliferations of SK-MEL-2 or B16F10. Futhermore, CUP showed inhibitory action against SK-MEL-2 proliferation at the concentration of $500{\mu}g/m{\ell}$ In addition, CUP was shown to have DPPH free radical scavenging activities and also have inhibitory effects on elastase activities too. On the fluorescent examinations, the present author knows that CUP elevated production levels of oxygen free radicals in malignant melanoma cell, SK-MEL-2. Conclusions : These results suggest that CUP has possibilities of usage for functional cosmetics which have wrinkle care and whitening activities and related mechanisms are involved in inhibition of elastase action and acceleration of oxidative stress in melanoma cell.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.2
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• pp.631-637
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• 2013

Luteolin is a naturally occurring flavonoid present in many plants with diverse applications in pharmacology. Despite several studies elucidating its significant anti-cancer activity against various cancer cells, the mechanism of action in skin cancer is not well addressed. Hence, we investigated the effects of luteolin in HaCaT (human immortalized keratinocytes) and A375 (human melanoma) cells. The radical scavenging abilities of luteolin were determined spectrophotometrically, prior to a cytotoxic study (XTT assay). Inhibitory effects were assessed by colony formation assay. Further, the capability of luteolin to induce cell cycle arrest and apoptosis were demonstrated by flow cytometry and cellular DNA fragmentation ELISA, respectively. The results revealed that luteolin possesses considerable cytotoxicity against both HaCaT and A375 cells with $IC_{50}$ values of 37.1 ${\mu}M$ and 115.1 ${\mu}M$, respectively. Luteolin also inhibited colony formation and induced apoptosis in a dose and time-dependent manner by disturbing cellular integrity as evident from morphological evaluation by Wright-Giemsa staining. Accumulation of cells in G2/M (0.83-8.14%) phase for HaCaT cells and G0/G1 (60.4-72.6%) phase for A375 cells after 24 h treatment indicated cell cycle arresting potential of this flavonoid. These data suggest that luteolin inhibits cell proliferation and promotes cell cycle arrest and apoptosis in skin cancer cells with possible involvement of programmed cell death, providing a substantial basis for it to be developed into a potent chemopreventive template for skin cancer.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.4
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• pp.329-335
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• 2017

We have studied on the keratinocytes differentiation and skin barrier function using Adenophorae radix (A. radix) root extract, which was known to contain triterpenoid, saponin and starch. A. radix root extracts showed the $PPAR{\alpha}$ expression level of Wy-14,643 $0.5-1.0{\mu}M$ in CV-1 cells. The cornified envelop formation (CE) of human keratinocyte cell line (HaCaT) and normal human keratinocyte (NHK) showed a statistically significant increased compared to the control. When HaCaT cells were treated with A. radix root extract, transglutaminase (TGase-1) was significantly increased. As a result of clinical study of the simple cosmetic formulation containing A. radix root extract for about 2 weeks, TEWL values were significantly decreased and water contents were increased. The ceramides, which were obtained from the inner forearm, were also significantly increased statistically. We suggest that the A. radix root extract can be used as a preventive and therapeutic agent for skin diseases such as dry skin and atopy.

• Journal of Ginseng Research
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• v.46 no.1
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• pp.115-125
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• 2022

Background: Ginsenosides (GS) have potential value as cosmetic additives for prevention of skin photoaging. However, their protective mechanisms against skin barrier damage and their active monomeric constituents are unknown. Methods: GS monomer types and their relative proportions were identified. A UVB-irradiated BALB/c hairless mouse model was used to assess protective effects of GS components on skin epidermal thickness and transepidermal water loss (TEWL). Skin barrier function, reflected by filaggrin (FLG), involucrin (IVL), claudin-1 (Cldn-1), and aquaporin 3 (AQP3) levels and MAPK phosphorylation patterns, were analyzed in UVB-irradiated hairless mice or HaCaT cells. Results: Total GS monomeric content detected by UPLC was 85.45% and was largely attributed to 17 main monomers that included Re (16.73%), Rd (13.36%), and Rg1 (13.38%). In hairless mice, GS ameliorated UVB-induced epidermal barrier dysfunction manifesting as increased epidermal thickness, increased TEWL, and decreased stratum corneum water content without weight change. Furthermore, GS treatment of UVB-irradiated mice restored protein expression levels and epidermal tissue distributions of FLG, IVL, Cldn-1, and AQP3, with consistent mRNA and protein expression results obtained in UVB-irradiated HaCaT cells (except for unchanging Cldn-1 expression). Mechanistically, GS inhibited JNK, p38, and ERK phosphorylation in UVB-irradiated HaCaT cells, with a mixture of Rg2, Rg3, Rk3, F2, Rd, and Rb3 providing the same protective MAPK pathway inhibition-associated upregulation of IVL and AQP3 expression as provided by intact GS treatment. Conclusion: GS protection against UVB-irradiated skin barrier damage depends on activities of six ginsenoside monomeric constituents that inhibit the MAPK signaling pathway.

• The Korea Journal of Herbology
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• v.38 no.5
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• pp.61-67
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• 2023

Objective : Mori Fructus has been used to relieve thirst, and treat dizziness, tinnitus, and insomnia caused by poor constitution. This study was performed to investigate the protective effect of the ethanol extract of Mori Fructus (MF) in ultraviolet B (UVB)-induced apoptotic cell death in human keratinocyte cells. Methods : MF was prepared by extracting 100 g of Mori Fructus in 1 L of 100% ethanol for 48 h. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to measure cell viability. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and the expression of apoptosis-related proteins was observed by Western blot. Tyrosinase activity was measured with a colorimetric commercial kit. Results : MF promoted cell vitality and inhibited apoptosis of UVB-induced HaCaT cells. MF pretreatment reduced TUNEL-positive cells and increased the expression of caspase-3 and -9. MF also displayed antioxidant effect with high radical scavenging ability. At 2 ㎎/㎖ concentration, the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical inhibition rates were 55.3 ± 4.6% and 48.5 ± 1.3%, respectively. Furthermore, MF showed a concentration-dependent inhibitory effect on tyrosinase. Conclusion : These results suggest that MF functions as a protective regulator in UVB-induced HaCaT cells by regulating apoptosis and partially exerting antioxidant effects. In addition, the tyrosinase inhibitory effect of MF shows the potential for MF to be used for skin pigmentation.

• Journal of Microbiology and Biotechnology
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• v.30 no.9
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• pp.1379-1386
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• 2020

Acne is a chronic skin disease that typically occurs in the teens and twenties, and its symptoms vary according to age, sex, diet, and lifestyle. The condition is characterized by hyperproliferation of keratinocytes in the epidermis, sebum overproduction, excessive growth of Propionibacterium acnes, and P. acnes-induced skin inflammation. Interleukin (IL)-1α and IL-6 are predominant in the inflammatory lesions of acne vulgaris. These cytokines induce an inflammatory reaction in the skin in the presence of pathogens or stresses. Moreover, IL-1α accelerates the production of keratin 16, which is typically expressed in wounded or aberrant skin, leading to abnormalities in architecture and hyperkeratinization. Orobol (3',4',5,7-tetrahydroxyisoflavone) is a metabolite of genistein that inhibited the P. acnes-induced increases in IL-6 and IL-1α levels in human keratinocytes (HaCaTs) more effectively compared with salicylic acid. In addition, orobol decreased the IL-1α and IL-6 mRNA levels and inhibited the phosphorylation of inhibitor of kappa-B kinase, nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha, and mitogen-activated protein kinase induced by P. acnes. Finally, the expression of Ki67 was decreased by orobol. Thus, orobol ameliorated the inflammation and hyperkeratinization induced by heat-killed P. acnes and thus has potential for use in functional foods and cosmetics.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.1013-1014
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• 2005