• 생명과학회지
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• 제29권10호
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• pp.1071-1079
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• 2019

본 연구에서 우리는 오미자 종자 분획물의 항산화 활성과 자외선에 유도된 human HaCaT cell에서의 MMPs의 발현억제에 의한 항노화 효과를 조사하고자 하였다. 전자공여능, ABTS 라디칼 소거분석, 그리고 hydrogen peroxide 소거분석 실험을 통해, 오미자 종자의 분획물 중 에틸아세테이트 분획물(SCEAf)이 가장 우수한 라디칼 소거활성을 가졌고, collagenase 저해 활성 실험에서 SCEAf는 $500{\mu}g/ml$의 농도에서 92.3% 이상의 저해효과를 보였음을 확인하였다. SCEAf의 HaCaT cell에 대한 세포 독성을 확인하기 위해 MTT assay를 실시한 결과, SCEAf는 세포에 독성이 없음을 나타낼 뿐 아니라 UVB에 손상된 세포의 생존율을 증가시킴으로써 세포 활성에 도움을 주는 것을 확인할 수 있었다. SCEAf의 HaCaT cell에서의 항노화 효과를 조사하기 위해 UVB $50mJ/cm^2$에 유도된 HaCaT cell에 SCEAf를 처리한 후 MMP-1과 -3의 발현을 Western blotting과 reverse-transcription polymerase chain reaction으로 분석하였다. 그 결과 SCEAf가 MMP-1과 -3의 단백질과 mRNAs의 발현을 농도의존적으로 억제시켰다. 이러한 결과는 오미자 종자 에틸아세테이트 분획물이 자외선의 손상을 받은 피부 각질형성세포에서의 collageanse 활성을 억제하여 노화를 예방하고 증상을 완화시킬 것으로 기대할 수 있다. 이러한 결과를 바탕으로 우리는 오미자 종자가 화장품과 식품 산업에서 항노화 효과가 있는 기능성 원료로서 사용하기 위한 잠재성을 가질 것으로 기대한다.

• KSBB Journal
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• 제28권6호
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• pp.394-399
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• 2013

Astragalus membranaceus Bunge is used in herbal medicine in Eastern Asian countries including Korea. In this study, we assessed the effects of A. membranaceus extract (AM) on the aquaporin-3 (AQP3) protein expression in HaCaT cells. AM did not affect viability of HaCaT cells. AQP3 expression and cell migration seem to be maximal at $100{\mu}g/mL$ concentration. Epidermal growth factor receptor (EGFR) kinase inhibitor, PD153035, blocked AM-induced AQP3 expression and cell migration. In addition, an 80% ethanol extracts of herbal prescription, SinhyoTakleesan (ST), which is composed of A. membranaceus, Angelicae gigantis, Glycyrrhiza glabra Linne, and Lonicera japonica Flos also induced AQP3 expression at $20{\mu}g/mL$ in HaCaT cells. Collectively, these results suggest that AM induce AQP3 expression via EGFR pathway.

• Korean Journal of Acupuncture
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• 제39권2호
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• pp.43-53
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• 2022

Objectives : Paeonia lactiflora Pallas (PLP) have been reported to have pharmacological effects such as anti-inflammatory and analgesic. However, it is not yet known whether PLP extract has anti-inflammatory effect on HaCaT cells, human keratinocyte. Methods : To confirm the anti-inflammatory effect of PLP on keratinocyte, TNF-𝛼/IFN-𝛾-stimulated HaCaT cells were used. HaCaT cells were pre-treated with PLP for 1h before stimulation with TNF-𝛼/IFN-𝛾. Then HaCaT cells were stimulated with TNF-𝛼/IFN-𝛾 for 24 h, the cells and media were harvested to measure the inflammatory cytokines levels. Granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), interleukin 1 beta (IL-1𝛽), and TNF-𝛼 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression of thymus and activation-regulated chemokines (TARC), IL-6, and IL-8 were measured by reverse transcription-polymerase chain reaction (RT-PCR). We also investigated the inhibitory mechanism of the mitogen-activated protein kinase (MAPKs) including ERK, JNK, and p38 and nuclear factor-kappaB (NF-𝜅B) by PLP using western blot. Results : PLP did not show cytotoxicity in HaCaT cells. In TNF-𝛼/IFN-𝛾-stimulated HaCaT cells, PLP significantly inhibited the expression of GM-CSF, MCP-1 IL-1𝛽, TNF-𝛼, TARC and IL-6. PLP inhibited the phosphorylation of ERK and translocation of NF-𝜅B into the nucleus. Conclusions : These results indicate that PLP could ameliorate the TNF-𝛼/IFN-𝛾-stimulated inflammatory response through inhibition of MAPK and NF-kB signal pathway. This suggests that PLP could be used beneficial agent to improve skin inflammation.

• 대한통합의학회지
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• 제12권1호
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• pp.63-71
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• 2024

Purpose : Skin is the primary barrier to protect the body from various exogenous factors. Among them, UVB exposure can cause the induction of not only excessive inflammatory responses but also the degradation of extracellular matrix (ECM), including collagen and elastin. This study tried to investigate the ameliorative effect of Persicaria perfoliata ethanol extract (PPEE) on UVB-irradiated photodamage through the regulation of activator protein (AP)-1, phosphoinositide 3-kinase (PI3K)/Akt, and mitogen-activated protein kinase (MAPK) signaling molecules in HaCaT cells. Methods : The cytotoxicity of PPEE on HaCaT cells was evaluated by the WST-1 assay. The 80 mJ/cm² of UVB (312 nm) was irradiated on HaCaT cells to induce the photodamage. Western blot analysis was conducted to investigate the protein expression levels of cyclooxygenase (COX)-2, matrix metalloproteinase (MMP)-9, and heme oxygenase (HO)-1 for ameliorative status by PPEE treatment in UVB-exposed HaCaT cells. In addition, the activated status of the inflammatory transcription factor, AP-1, as well as upstream signaling molecules, PI3K/Akt, and MAPK, were also evaluated by Western blot analysis. Results : Any cytotoxic effect was not induced at the concentration up to 200 ㎍/ml by PPEE treatment. Protein expression levels of COX-2 and MMP-9 were significantly down- and up-regulated by PPEE treatment. The inflammatory transcription factor AP-1, stimulated by UVB irradiation, was also significantly attenuated by PPEE treatment. The phosphorylated status of PI3K/Akt and MAPK were mitigated by PPEE treatment in UVB-exposed HaCaT cells. Moreover, PPEE treatment potently accelerated the expression of HO-1 and its transcription factor, nuclear factor-erythroid 2-related factor (Nrf)2, which is known for its anti-inflammatory activity. Conclusion : Consequently, PPEE treatment significantly regulated COX-2 and MMP-9 expressions in UVB-irradiated HaCaT cells. The inflammatory transcription factor AP-1, along with upstream signaling molecules PI3K/Akt and MAPKs, were also attenuated by PPEE treatment in UVB-exposed HaCaT cells. Additionally, PPEE treatment exaggerated HO-1 expression and Nrf2 activation, which might have contributed to the anti-inflammatory activity of PPEE. These results indicate that PPEE could be a candidate for attenuating UVB-induced photodamage in human skin.

• Journal of Ginseng Research
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• 제42권2호
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• pp.229-232
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• 2018

• 한국수산과학회지
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• 제49권5호
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• pp.589-593
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• 2016

Propionibacterium acnes infection in skin tissue often causes acne vulgaris, commonly characterized by inflammatory papules, pustules, and nodules. Chitosan and its derivatives possess strong anti-inflammatory effects. In this study, the anti-inflammatory activity of chitosan-phytochemical conjugates on P. acnes-infected human skin keratinocytes (HaCaT) was evaluated. We designed a model of P. acnes-induced inflammation in viable HaCaT cells. Nitric oxide (NO), an inflammatory marker, was successfully elevated by P. acnes infection in HaCaT cells in a dose-dependent manner. Furthermore, the levels of NO were reduced by treatment with chitosan-phytochemical conjugates (chitosan-caffeic acid, -ferulic acid and -sinapic acid) in a dose-dependent manner. Among these conjugates, chitosan-caffeic acid exhibited the strongest NO suppression in HaCaT cells infected with P. acnes. The results obtained in this study suggest that chitosan-phytochemical conjugates could be used as a potential therapeutic agent against acne vulgaris.

• 생명과학회지
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• 제27권3호
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• pp.310-317
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• 2017

본 연구진은 HaCaT-ARE-luciferase 세포를 이용하여 400 여개의 약용식물 에탄올 추출물 중 NRF2/ARE 유도효과가 있는 신규 추출물을 검색하였고 이를 통하여 종대황(Rheum undulatum)과 선복화(Inula japonica)의 주정 추출물이 HaCaT-ARE-luciferase 세포에서 ARE 활성을 강하게 유도하는 것을 관찰하였다. 종대황과 선복화 에탄올 추출물은 HaCaT 세포에서 생존(viability)을 증가시켰고 NRF2/ARE에 의존적인 phase II cytoprotective 효소인 heme oxygenase-1 (HO-1)와 NADPH:quinone oxidoreductase-1 (NQO1)의 전사 및 단백질 발현을 강하게 유도하였다. 또한 종대황과 선복화 추출물은 HaCaT 세포에서 TPA로 유도한 세포 내 활성 산소 및 이를 통하여 생성되는 스트레스 마커인 8-hydroxydeoxyguanosine (8-OH-dG)과 4-hydroxynonenal (4HNE)의 발생을 강하게 억제하였다. 본 연구는 종대황과 선복화의 에탄올 추출물이 인간 피부 세포주인 HaCaT 세포에서 NRF2/ARE에 의존적인 유전자 발현의 유도를 통하여 강력한 항산화 효과를 발휘한다는 것을 증명한다.

• 생약학회지
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• 제45권3호
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• pp.256-261
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• 2014

Bee venom (BV) therapy has been used as a traditional medicine to treat a variety of conditions, such as arthritis, back pain, cancerous tumors, and skin diseases. However, regulatory effects of BV on tumor necrosis factor (TNF)-${\alpha}$-induced HaCaT cell migration or anti-inflammatory have not been explored. In the present study, we investigated the effects of BV on HaCaT cell migration and anti-inflammation. HaCaT cell migration was evaluated by wound-healing assay. The pro-inflammatory cytokines such as TNF-${\alpha}$, interleukin (IL)-$1{\beta}$, and IL-8 were examined by ELISA or Western blotting. BV treatment led to an increase in migration of HaCaT cells for 24 and 48 h. Especially, 10 ng/ml of BV were significantly increased HaCaT cell migration. Also, BV suppressed the secretion of TNF-${\alpha}$, IL-$1{\beta}$, and IL-8 in culture medium with HaCaT cells. In addition, Western blot results demonstrate that BV suppressed the expression of TNF-${\alpha}$ and IL-$1{\beta}$, in HaCaT cells. Especially, 1 or 10 ng/ml of BV markedly decreased the expression of pro-inflammatory cytokines. These results demonstrate the potential of BV for the prevention of skin inflammation induced by TNF-${\alpha}$.

• Asian Pacific Journal of Cancer Prevention
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• 제13권5호
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• pp.2015-2020
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• 2012

Oleanolic acid (OA) is a naturally occurring triterpenoid in food materials and is a component of the leaves and roots of Olea europaea, Viscum album L., Aralia chinensis L. and more than 120 other plant species. There are several reports validating its antitumor activity against different cancer cells apart from its hepatoprotective activity. However, antitumor activity against skin cancer has not beed studied well thus far. Hence the present study of effects of OA against HaCaT (immortalized keratinocyte) cells - a cell-based epithelial model system for toxicity/ethnopharmacology-based studies - was conducted. Radical scavenging activity ($DPPH{\cdot}$) and FRAP were determined spectrophotometrically. Proliferation was assessed by XTT assay at 24, 48 and 72 hrs with exposure to various concentrations (12.5-200 ${\mu}M$) of OA. Apoptotic induction potential of OA was demonstrated using a cellular DNA fragmentation ELISA method. Morphological studies were also carried out to elucidate its antitumor potential. The results revealed that OA induces apoptosis by altering cellular morphology as well as DNA integrity in HaCaT cells in a dose-dependent manner, with comparatively low cytotoxicity. The moderate toxicity observed in HaCaT cells, with induction of apoptosis, possibly suggests greater involvement of programmed-cell death-mediated mechanisms. We conclude that OA has relatively low toxicity and has the potential to induce apoptosis in HaCaT cells and hence provides a substantial and sound scientific basis for further validation studies.

• 생약학회지
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• 제46권2호
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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.