• Title/Summary/Keyword: HaCaT keratinocyte

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Antioxidative and Protective Effects of Corn Silk (Zea mays L.) Extract on Human HaCaT Keratinocyte (옥수수수염 추출물의 항산화효과 및 피부각질세포 보호효과)

  • Kim, Hyun Young;Seo, Woo Duck;Seo, Kyung Hye;Lee, Mi-Ja;Choi, Sik-Won;Lee, Kwang-Sik;Kim, Sun Lim;Kang, Hyeon Jung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.61 no.3
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    • pp.184-190
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    • 2016
  • We investigated the antioxidative and protective effects of corn silk (Zea mays L.) ethanol extracts on human HaCaT cells and erythrocytes. The NICS-2 fraction, extracted from corn silk, exhibited favorable 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activities with $IC_{50}$valuesof$13.3{\pm}0.3{\mu}g/mL$ and $14.2{\pm}0.1{\mu}g/mL$ when compared with those of ${\alpha}$-tocopherol, a positive control, with $IC_{50}=10.4{\pm}02.2$ and $22.2{\pm}3.6{\mu}g/mL$, respectively. In addition, we investigated skin protection effects of NICS extracts of corn silk in HaCaT keratinocytes. To investigate the pharmacological potential of NICS-1 and NICS-2 extracts of corn silk on UV-B-induced damage in HaCaT cells, we measured the activity of interleukin (IL) 1a. Our results showed that all the corn silk extracts inhibited the UV-B-induced activity of IL-1a. In particular, NICS-1 extracts of corn silk significantly suppressed IL-1a activity in a dose-dependent manner without inducing cytotoxicity. These results indicate that the ethanol extracts of corn silk (Zea mays L.) could function as natural cytoprotective agents and antioxidants in biological systems, particularly the skin exposed to UV radiation, by protecting cellular membrane against reactive oxygen species (ROS).

The Effects of the Fruits of Foeniculum vulgare on Skin Barrier Function and Hyaluronic Acid Production in HaCaT Keratinocytes (HaCaT 세포에서 회향 열매의 피부장벽기능과 hyaluronic acid 생성에 미치는 영향)

  • Yu, Hak Yin;Yang, In Jun;Lincha, V.R;Park, In Sik;Lee, Dong-Ung;Shin, Heung Mook
    • Journal of Life Science
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    • v.25 no.8
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    • pp.880-888
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    • 2015
  • Foeniculum vulgare (FV) has long been used in traditional medicine for the treatment of inflammatory diseases. In addition, it is usually known as an important medicinal and aromatic plant widely used as a carminative, digestive, lactogogue, and diuretic, and for treating respiratory and gastrointestinal disorders. The skin barrier protects against the invasion of pathogens, fends off chemical and physical assaults, and protects against extensive water loss. In this study, the effects of solvent-fractionated FV fruits on strengthening the skin barrier and maintaining moisture, as well as their antifungal activity, were investigated in human keratinocyte (HaCaT) cells. The expression of involucrin, loricrin, filaggrin, hyaluronic acid synthase, human β defensin, and cathelicidin genes and proteins was measured by reverse transcription polymerase chain reaction (RT-PCR) and western blotting. The production of hyaluronic acid was determined by enzyme-linked immunosorbent assay (ELISA). The butanol fraction increased the expression of involucrin and filaggrin. Both the ethyl acetate and the butanol fractions increased hyaluronic acid production by promoting the expression of hyaluronic acid synthase-1. Although the antimicrobial peptides were increased by FV crude extract and its fractions, the samples did not show a significant effect compared to the normal group. These results suggest that the butanol fraction of FV could be very useful in cosmetics for the treatment of dermatological diseases.

Suppression of TNF-α-induced Inflammation by Extract from Different Parts of Moringa in HaCaT Cells (HaCaT 각질형성세포에서 TNF-α에 의하여 유도되는 염증 발현에 대한 부위별 모링가 추출물의 억제 효과)

  • Lee, Hyo-Jin;Chang, Young-Chae
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1254-1260
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    • 2012
  • The moringa (Moringa oleifera Lam.) plant is used both as food and an anti-allergic agent. In this study, we investigated skin protection effects of methanol extracts from the root, seed, fruit, and leaves of moringa in HaCaT keratinocyte cells. To investigate the pharmacological potential of various moringa extracts on TNF-${\alpha}$-induced collagen degradation in HaCaT cells, we measured the activity of matrix metallopeptidase-9,2 (MMP-9,2) by zymography analysis. Our results showed that all the moringa extracts inhibit the TNF-${\alpha}$-induced enzyme activity of MMP-9. In particular, moringa root extracts significantly suppressed MMP-9 and MMP-2 in a dose-dependent manner. Next, to investigate the anti-inflammation effect of the moringa extracts, we examined cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and interleukin-6 (IL-6) expression of the extracts. The results showed that both the root extracts and the seed extracts decreased the TNF-${\alpha}$-induced expression of COX-2. In addition, the root and leaf extracts reduced the expression of IL-6. However, none of the moringa extracts affected the expression of iNOS. The results suggest that moringa root extracts down-regulate MMP-9, COX-2, and IL-6 and that the root extracts offer superior skin protection effects compared with other extracts of moringa in HaCaT cells.

Quercetin suppress CCL20 by reducing IκBα/STAT3 phosphorylation in TNF-α/IL-17A induced HaCaT cells (TNF-α/IL-17A 유도된 HaCaT 세포주에서 Quercetin의 IκBα/STAT3 인산화 조절에 의한 CCL20 발현 억제)

  • Kim, Mi Ran;Kim, Min Young;Hwang, Hyung Seo
    • Journal of Applied Biological Chemistry
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    • v.63 no.3
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    • pp.211-219
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    • 2020
  • Quercetin is a polyphenol compound with excellent antioxidant and anti-inflammatory activity. However, little has been reported about the efficacy of quercetin to control psoriasis. Thus, we aimed to investigate the effect of quercetin to regulate psoriatic dermatitis with HaCaT cell lines activated by TNF-α and IL-17A, which are in vitro psoriasis skin models. When quercetin was treated with TNF-α-activated HaCaT cell line, inflammatory cytokine expressions such as IL-1α, IL-1β and IL-6 were reduced by 49.1±7.14, 42.8±8.16, and 34.5±2.52%, respectively. In addition, mRNA expression levels of IL-8 and CCL20 the chemokines that attract immune cells such as Th17 cells and dendritic cells to the inflammatory reaction site, were also reduced by 38.4±5.83 and 52.9±4.59% compared to the TNF-α treatment group. The expression of proteins KRT6A and KRT16, which was nonspecifically increased in psoriatic skin was also significantly suppressed. Moreover, phosphorylation of IκBα and STAT3 proteins activated by TNF-α was also significantly inhibited. After stimulating the HaCaT with IL-17A, known as another psoriasis-inducing cytokine, it was observed that IκBα mRNA expression decreased by 55.8±5.28%, and STAT3 phosphorylation was downregulated by 36.3±6.81%. Finally, after co-activation by TNF-α/IL-17A, quercetin inhibited all of IL-1α, IL-1β, IL-6, TNF-α and CCL20 gene expression. The above results strongly suggest that quercetin is a material that has not only anti-oxidant and anti-inflammatory activities, but also has an activity in improving psoriasis.

Brazilin downregulates CCL20 expression via regulation of STAT3 phosphorylation in TNF-α/IL-17A/IFN-γ-induced HaCaT cells (TNF-α/IL-17A/IFN-γ 유도된 HaCaT 세포에서 브라질린의 STAT3 인산화 억제를 통한 CCL20 저해 효과)

  • Kim, Mi Ran;Hwang, Hyung Seo
    • Journal of Applied Biological Chemistry
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    • v.64 no.2
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    • pp.185-192
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    • 2021
  • Psoriasis is a chronic intractable skin disease caused by various inflammatory cytokines such as IL-6, CXCL8, TNF-α, and IFN-γ, as well as IL-17A secreted from Th17 cells and is characterized by hyperkeratosis and chronic inflammation of the epidermis. Brazilin, an active ingredient of Caesalpinia sappan L., is known to exert antioxidant and anti-inflammatory activity, and function in skin barrier improvement. In particular, it was shown as a potential material for treating psoriasis in a tumor necrosis factor (TNF)-α-stimulated HaCaT keratinocyte model. However, the direct regulation of the C-C motif chemokine ligand (CCL) 20, a psoriasis-inducing factor, by brazilin has not been reported. Therefore, in this study, we investigated the suppression of CCL20 and the regulatory mechanism by brazilin using a psoriasis-like model. First, brazilin downregulated CCL20 and CXCL8 in IL-17A-stimulated HaCaT cells in a concentration-dependent manner by inhibiting signal transducer and transcription (STAT)3 phosphorylation. In addition, brazilin significantly inhibited the expression of psoriasis-related genes CXCL8, CCL20, IL-1, IL-6, and TNF-α in TNF-α/IL-17A/IFN-γ-stimulated HaCaT cells. Moreover, brazilin also had a positive effect on improving the skin barrier in TNF-α/IL-17A/IFN-γ-stimulated HaCaT cells. The above results indicated that brazilin ultimately downregulated CCL20 expression by inhibiting STAT3 phosphorylation, and also suppressed the expression of psoriasis-induced cytokines. If the efficacy of brazilin in improving psoriasis is verified through animal models and clinical trials in the future, it may represent a potentially therapeutic substance for psoriasis patients.

Simultaneous Analysis and Anti-allergic Effect of Mume Fructus (오매(烏梅)의 다성분 동시분석 및 항알러지 효과)

  • Seo, Chang-Seob;Ha, Hyekyung;Lee, Ho Young;Lee, JunKyung;Jung, Dayoung;Lee, Jin-Ah;Shin, Hyeun Kyoo
    • Korean Journal of Pharmacognosy
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    • v.43 no.4
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    • pp.279-285
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    • 2012
  • The Mume Fructus (MF) has been used for relieves cough, arrests arrest chronic diarrhea, treat fluid depletion, and treat ascariasis in Korea. In this study, a high-performance liquid chromatography (HPLC) method was established for simultaneous determination of six main components of MF. Additionally, we were investigated the anti-inflammatory and anti-allergic effects of MF extract on lipopolysaccharide (LPS)-treated RAW264.7 cells and tumor necrosis factor (TNF)-${\alpha}$/interferon (IFN)-${\gamma}$-treated HaCaT cells. The analytical column for separation was used a Gemini $C_{18}$ column maintained at $40^{\circ}C$. The mobile phase consisted of 1.0% (v/v) acetic acid in water (A) and 1.0% (v/v) acetic acid in acetonitrile (B). The flow rate was 1.0 mL/min and the detector was a photodiode array (PDA) set at 280 nm and 320 nm. We evaluated the inhibitory effect of MF extract on the production of inflammatory markers, nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) in LPS-stimulated RAW264.7 cells and thymus- and activation-regulated chemokine (TARC/CCL17) in TNF-${\alpha}$/IFN-${\gamma}$-treated HaCaT cells, respectively. We confirmed the genes expression related with TARC, macrophage-derived chemokine (MDC/CCL22) and regulated on activation, normal T cell expressed and secreted (RANTES/CCL5) in HaCaT keratinocyte cells by MF extract. The contents of the five compounds in MF were 0.22-1.01 mg/g. Also, the MF extract show inhibition of about 78% and 75% on NO and $PGE_2$ production at the concentration 1000 mg/mL in RAW264.7 cells. MF extract suppressed the hTARC level and genes expression such as TARC, MDC, and RANTES on TNF-${\alpha}$/IFN-${\gamma}$-treated HaCaT cells.

The Protective Effects of Astragali Radix Against UV Induced Cellular Damage in Human Keratinocytes (황기의 자외선에 의한 세포 손상을 막는 보호 효과)

  • Lee, Jin-Young;Park, Hye-Yoon;Yeom, Myeong-Hun;Kim, Duck-Hee;Kim, Han-Kon
    • Korean Journal of Pharmacognosy
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    • v.39 no.4
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    • pp.300-304
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    • 2008
  • The root of Astragalus membranaceus Bunge (Leguminosae) has been used in the Korean oriental medicine for strengthening the vital energy. UV irradiation has been suggested as a major cause of photo aging in skin. In order to investigate protective effects against UV induced cellular damage, Astragali Radix was extracted with 70% ethanol and dissolved in DMSO. The protective effect was detected by MTT assay, LDH assay, and Comet assay in immortalized human keratinocyte cell line, HaCaT cell system after UV irradiation. Astragli Radix 70% EtOH extract reduced UV induced cellular damage in cell survival, membrane integrity and DNA damage.

Skin Barrier Recovery by Protease-Activated Receptor-2 Antagonist Lobaric Acid

  • Joo, Yeon Ah;Chung, Hyunjin;Yoon, Sohyun;Park, Jong Il;Lee, Ji Eun;Myung, Cheol Hwan;Hwang, Jae Sung
    • Biomolecules & Therapeutics
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    • v.24 no.5
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    • pp.529-535
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    • 2016
  • Atopic dermatitis (AD) results from gene and environment interactions that lead to a range of immunological abnormalities and breakdown of the skin barrier. Protease-activated receptor 2 (PAR2) belongs to a family of G-protein coupled receptors and is expressed in suprabasal layers of the epidermis. PAR2 is activated by both trypsin and a specific agonist peptide, SLIGKV-$NH_2$ and is involved in both epidermal permeability barrier homeostasis and epithelial inflammation. In this study, we investigated the effect of lobaric acid on inflammation, keratinocyte differentiation, and recovery of the skin barrier in hairless mice. Lobaric acid blocked trypsin-induced and SLIGKV-$NH_2$-induced PAR2 activation resulting in decreased mobilization of intracellular $Ca^{2+}$ in HaCaT keratinocytes. Lobaric acid reduced expression of interleukin-8 induced by SLIGKV-$NH_2$ and thymus and activation regulated chemokine (TARC) induced by tumor necrosis factor-a (TNF-${\alpha}$) and IFN-${\gamma}$ in HaCaT keratinocytes. Lobaric acid also blocked SLIGKV-$NH_2$-induced activation of ERK, which is a downstream signal of PAR2 in normal human keratinocytes (NHEKs). Treatment with SLIGKV-$NH_2$ downregulated expression of involucrin, a differentiation marker protein in HaCaT keratinocytes, and upregulated expression of involucrin, transglutamase1 and filaggrin in NHEKs. However, lobaric acid antagonized the effect of SLIGKV-$NH_2$ in HaCaT keratinocytes and NHEKs. Topical application of lobaric acid accelerated barrier recovery kinetics in a SKH-1 hairless mouse model. These results suggested that lobaric acid is a PAR2 antagonist and could be a possible therapeutic agent for atopic dermatitis.

Comparative Study of the Anti-inflammatory Effects of Menthae Herba from Korea and China (한국산과 중국산 박하의 항염증 효과에 관한 비교연구)

  • Lim, Hye-Sun;Kim, Jung-Hoon;Ha, Hye-Kyung;Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.43 no.3
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    • pp.231-238
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    • 2012
  • Menthae herba (MH) extracts exhibit anti-inflammatory effects. The purpose of this study was to determine whether the anti-inflammatory effects of MH extracts vary according to the cultivation regions. We performed a comparative analysis of MH extracts by evaluating the production of inflammatory mediators in RAW 264.7 murine macrophage cells and HaCaT human keratinocyte cells. MH extracts obtained from different cultivation regions in Korea and China significantly reduced the production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in RAW 264.7 cells stimulated with lipopolysaccharide (LPS). No differences in these inhibitory activities were observed between MH extracts. In HaCaT cells stimulated with TNF-${\alpha}$ and interferon-${\gamma}$ (IFN-${\gamma}$), MH extracts did not inhibit the production of macrophage-derived chemokine (MDC/CCL22), but most extracts reduced the production of the regulated on activation normal T-cell expression and secreted (RANTES/CCL5). We used clustering tree analysis of the MH extracts according to the chromatographic pattern and anti-inflammatory potency of MH extracts. We observed differences in the chromatographic pattern of MH extracts but no difference in anti-inflammatory potency. Our findings suggest that MH extracts from different regions do not show any differences in their pharmacological potency in that MH extracts are used as therapeutic agents to treat inflammatory disorders.

Preventive Effect of Natural Pigments Against Ultraviolet B-induced Cell Death in HaCat Cells

  • Lim, Jae-Chung;Bae, Chun-Sik;Jeong, Soo-Young;Boo, Hee-Ock;Hwang, Seong-Jin;Lim, Seul-Ki;Park, Min-Jung;Kim, Jong-Chun;Kang, Seong-Soo;Han, Ho-Jae;Park, Soo-Hyun
    • Biomedical Science Letters
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    • v.17 no.1
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    • pp.55-60
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    • 2011
  • Skin is a physical barrier against diverse injury and damages. Exposure to ultraviolet (UV) radiation causes detrimental skin injuries such as inflammation and cell death. The value of natural pigments could be applied to many usages including cosmetics. This study was conducted to examine the protective effect of natural pigments extracted from mulberry, balsam pear, purple-colored sweet potato, pehmannia root, gardenia fruit, and black rice against UV-induced cell death in HaCaT cells, human keratinocyte cell lines. In the present study, the exposure of 50 mJ/$cm^2$ UV-B for 24 hr induced cell death in HaCaT cells, which was prevented by the pretreatment of extracts of mulberry, balsam pear, purple-colored sweet potato, rehmannia root, gardenia fruit, and black rice. In addition, the exposure of 50 mJ/$cm^2$ UV-B for 24 hr also increased lipid peroxide (LPO) formation, compared to control in HaCaT cells, which was prevented by the pretreatment of extracts of mulberry, balsam pear, purple-colored sweet potato, rehmannia root, gardenia fruit, and black rice. In conclusion, the extracts of mulberry, balsam pear, purple-colored sweet potato, rehmannia root, gardenia fruit, and black rice prevented the UV-B-induced cell apoptosis via the inhibition of oxidative stress in HaCaT cells.