• Title/Summary/Keyword: HaCaT keratinocytes

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Study on Skin pH Improvement Effect through Regulation of Na+/H+ Exchanger 1 (NHE1) Expression of Prunella vulgaris Extract and Its Active Compound, Caffeic Acid (꿀풀 추출물과 그 활성 화합물인 카페인산의 Na+/H+ exchanger 1 (NHE1) 발현 조절을 통한 피부 pH 개선 효과에 대한 연구)

  • No-June Park;Sim-Kyu Bong;Sang-A Park;Gi Hyun Park;Young Chul Ko;Hae Won Kim;Su-Nam Kim
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.1
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    • pp.87-96
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    • 2023
  • This study was conducted to discover substances that regulate skin surface acidification using human epidermal keratinocyte cell lines, and to investigate their effects on the moisturizing ability and skin barrier function of the stratum corneum. Prunella vulgaris (P. vulgaris) is an herb widely distributed in Northwest Africa and North America that has been studied for its anti-apoptotic, antioxidant, and anti-inflammatory effects. However, research on the regulation of NHE1 expression and the restoration of skin barrier function has not been conducted. Analysis of P. vulgaris revealed the presence of rosmarinic acid and caffeic acid as active ingredients, which were tested for toxicity in human epidermal keratinocyte cell lines (HaCaT), and showed no toxic effects were observed at high concentarion (100 ㎍/mL or 100 µM). It is known that sodium-hydrogen ion exchange pumps (NHE1) decrease in expression in aging skin to maintain the acidic pH of the stratum corneum, and it is hypothesized that this decrease plays an important role in the impaired restoration of skin barrier function in aging skin. P. vulgaris extract and caffeic acid increased the expression of NHE1 in keratinocytes, increased the expression of natural moisturizing factor (NMF) precursor filaggrin and ceramide synthesis enzyme serine palmitoyl transferase (SPT). In addition, P. vulgaris and caffeic acid decreased the extracellular pH of keratinocytes, indicating a direct effect on skin pH regulation. Taken together, these results suggest that P. vulgaris and caffeic acid can regulate skin pH through NHE1 modulation, and may help to restore skin barrier function by increasing NMF and ceramide synthesis. These results show the possibility that honeysuckle and caffeic acid can have a positive effect on skin health, and can be the basis for the development of new skin protection products using them.

Mechanisms of Suppression of Matrix Metalloproteinases in UVB-Irradiated HaCaT Keratinocytes of Colored Rice Varieties (UVB에 조사된 HaCaT Keratinocytes에서의 유색미에 의한 Matrix Metalloproteinases 발현억제 메커니즘)

  • Choi, Eun-Young;Lee, Jae-Bong;Kim, Do-Hoon;Kwon, Yong-Sham;Cheon, Jung-Yoon;Lee, Jin-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.5
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    • pp.562-571
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    • 2017
  • In this study, we investigated the anti-oxidant activities [electron-donating ability (EDA), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging ability, and reactive oxygen species (ROS) inhibitory activity], anti-wrinkle activities [collagenase inhibitory activity, suppression and/or phosphorylation of matrix metalloproteinases (MMPs), and mitogen-activated protein (MAP) kinase activity], and mRNA expression levels using reverse transcription-polymerase chain reaction (RT-PCR) assay in ultraviolet (UV) B ray ($50mJ/cm^2$)-irradiated human keratinocyte HaCaT cells. Josaengheugchal, Sinneungheugchal (SE), Shintoheug rice, Heugjinju rice, and Heugseol (HE) among colored rice varieties were reported to have excellent antioxidant properties. In the EDA and ABTS radical scavenging assays, extracts of the five colored rice varieties had scavenging activities of 72% at concentrations higher $50{\mu}g/mL$. In the collagenase inhibition assay, ethanol extracts of the five colored rice varieties showed high inhibitory effects of about 60% at concentrations higher $25{\mu}g/mL$. In the ROS inhibition assay, ethanol extracts of HE and SE showed very excellent inhibition efficacies at all concentrations. We determined molecular biological mechanisms of MMPs (MMP-1, -3, -8, and -13) and mitogen-activated protein kinase (MAPK) with HE, and the results show that HE suppressed expression of MMPs and phosphorylation of MAPK and increased expression of pro-collagen type I in UVB-irradiated cells. It was also confirmed by RT-PCR that HE reduced expression of MMPs mRNA. Therefore, these results suggest that HE has anti-wrinkle and collagen production effects and may be used as a material in the development of functional food and cosmetic industries.

Effects of water and ethanol extracts from Ojeok-san on inflammation and its related diseases (오적산의 추출 용매 차이에 따른 염증 및 염증 관련 질환에 대한 효능 비교)

  • Yoo, Sae-Rom;Jeong, Soo-Jin;Kim, Ye-Ji;Lim, Hye-Sun;Jin, Seong-Eun;Jeon, Woo-Young;Shin, In-Sik;Shin, Na-Ra;Kim, Seong-Sil;Kim, Jung-Hoon;Ha, Hye-Kyung;Lee, Mee-Young;Kim, Ohn-Soon;Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • The Journal of Internal Korean Medicine
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    • v.33 no.4
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    • pp.418-428
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    • 2012
  • Objectives : Ojeok-san, a traditional herbal formula, has been used for the treatment of cold illness and its related symptoms such as headache, nausea and indigestion. This study was performed to compare effects of water (OJSW) and 70% ethanol extracts (OJSE) of Ojeok-san on inflammation and its related diseases atopy, asthma and obesity in vitro. Methods : We performed HPLC to investigate contents of index components of OJSW and OJSE. We investigated the effects of OJSW and OJSE with an in vitro model, using 5 cell lines, specifically RAW 264. 7, HaCaT, MC/9, BEAS-2B and 3T3-L1. Results : HPLC analysis displayed that the contents of index components were higher in OJSE than OJSW. In lipopolysaccharide (LPS)-treated RAW 264.7 macrophages, OJSE significantly inhibited productions of interleukin (IL)-6, nitrite and prostaglandin $E_2$ ($PEG_2$). In TNF-${\alpha}$/IFN-${\gamma}$-treated HaCaT keratinocytes, OJSE significantly lowered levels of macrophage-derived chemokine (MDC) as well as regulated and normal T cell expressed and secreted (RANTES). OJSE also had a protective effect on inflammatory response by decreasing RANTES secretion in TNF-${\alpha}$-stimulated BEAS-2B cells. Conclusions : We conclude that OJSE could be more appropriate to enhance the biological activities against inflammation and its related diseases, and could be applied as a bioactive material for developing the potent anti-inflammatory agents.

Mechanism Underlying a Proteasome Inhibitor, Lactacystin-Induced Apoptosis on SCC25 Human Tongue Squamous Cell Carcinoma Cells (사람혀편평상피세포암종세포에서 proteasome 억제제인 lactacystin에 의해 유도된 세포자멸사의 기전에 대한 연구)

  • Baek, Chul-Jung;Kim, Gyoo-Cheon;Kim, In-Ryoung;Lee, Seung-Eun;Kwak, Hyun-Ho;Park, Bong-Soo;Tae, Il-Ho;Ko, Myung-Yun;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.34 no.3
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    • pp.261-276
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    • 2009
  • Lactacystin, a microbial natural product synthesized by Streptomyces, has been commonly used as a selective proteasome inhibitor in many studies. Proteasome inhibitors is known to be preventing the proliferation of cancer cells in vivo as well as in vitro. Furthermore, proteasome inhibitors, as single or combined with other anticancer agents, are suggested as a new class of potential anticancer agents. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in SCC25 human tongue sqaumous cell carcinoma cell line treated with lactacystin. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingiva fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay respectively. The hoechst staining, hemacolor staining and TUNEL staining were conducted to observe SCC25 cells undergoing apoptosis. SCC25 cells were treated with lactacystin, and Western blotting, immunocytochemistry, confocal microscopy, FAScan flow cytometry, MMP activity, and proteasome activity were performed. Lactacystin treatment of SCC25 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Interestingly, lactacytin remarkably revealed cytotoxicity in SCC25 cells but not normal cells. And tested SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, the up-regulation of Bax, and the activation of caspase-7, caspase-3, PARP, lamin A/C and DFF45 (ICAD). Flow cytometric analysis revealed that lactacystin resulted in G1 arrest in cell cycle progression which was associated with up-regulation in the protein expression of CDK inhibitors, $p21^{WAF1/CIP1}$ and $p27^{KIP1}$. We presented data indicating that lactacystin induces G1 cell cycle arrest and apoptois via proteasome, mitochondria and caspase pathway in SCC25 cells. Therefore our data provide the possibility that lactacystin could be as a novel therapeutic strategy for human tongue squamous cell carcinoma.

Levosulpiride, (S)-(-)-5-Aminosulfonyl-N-[(1-ethyl-2-pyrrolidinyl) methyl]-2-methoxybenzamide, enhances the transduction efficiency of PEP-1-ribosomal protein S3 in vitro and in vivo

  • Ahn, Eun-Hee;Kim, Dae-Won;Kim, Duk-Soo;Woo, Su-Jung;Kim, Hye-Ri;Kim, Joon;Lim, Soon-Sung;Kang, Tae-Cheon;Kim, Dong-Joon;Suk, Ki-Tae;Park, Jin-Seu;Luo, Qiuxiang;Eum, Won-Sik;Hwang, Hyun-Sook;Choi, Soo-Young
    • BMB Reports
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    • v.44 no.5
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    • pp.329-334
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    • 2011
  • Many proteins with poor transduction efficiency were reported to be delivered to cells by fusion with protein transduction domains (PTDs). In this study, we investigated the effect of levosulpiride on the transduction of PEP-1 ribosomal protein S3 (PEP-1-rpS3), and examined its influence on the stimulation of the therapeutic properties of PEP-1-rpS3. PEP-1-rpS3 transduction into HaCaT human keratinocytes and mouse skin was stimulated by levosulpiride in a manner that did not directly affect the cell viability. Following 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in mice, levosulpiride alone was ineffective in reducing TPA-induced edema and in inhibiting the elevated productions of inflammatory mediators and cytokines, such as cyclooxygenase-2, inducible nitric oxide synthase, interleukin-6 and -1${\beta}$, and tumor necrosis factor-${\alpha}$. Anti-inflammatory activity by PEP-1-rpS3 + levosulpiride was significantly more potent than by PEP-1-rpS3 alone. These results suggest that levosulpiride may be useful for enhancing the therapeutic effect of PEP-1-rpS3 against various inflammatory diseases.

Mechanism underlying Chios gum mastic-induced apoptosis on SCC25 human tongue squamous cell carcinoma cell line

  • Lee, Seung-Eun;Hur, Young-Joo;Kim, In-Ryoung;Kwak, Hyun-Ho;Kim, Gyoo-Cheon;Shin, Sang-Hun;Kim, Chul-Hoon;Park, Bong-Soo
    • International Journal of Oral Biology
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    • v.34 no.2
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    • pp.61-72
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    • 2009
  • Chios gum mastic (CGM) is a resin produced from the stem and leaves of Pistiacia lentiscus L var chia, a plant which grows only on Chios Island in Greece. CGM has been used for many centuries as a dietary supplement and folk medicine for stomach and duodenal ulcers in many Mediterranean countries and is known also to induce cell cycle arrest and apoptosis in some cancer cells. In this study, we further investigated the induction and mechanisms underlying the apoptotic response to CGM treatment in the SCC25 human tongue squamous cell carcinoma cell line. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingival fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay, respectively. Staining with Hoechst and hemacolor dyes and TUNEL assays were employed to detect SCC25 cells undergoing apoptosis. SCC25 cells were treated with CGM, and this was followed by western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, MMP activity and proteasome activity analyses. CGM treatment of SCC25 cells was found to result in a time- and dosedependent decrease in cell viability, a dose-dependent inhibition of cell growth, and apoptotic cell death. Interestingly, CGM showed a remarkable level of cytotoxicity in SCC25 cells but not in normal cells. Tested SCC25 cells also showed several lines of apoptotic manifestation. Taken together, our present findings demonstrate that CGM strongly inhibits cell proliferation by modulating the expression of G1 cell cycle-related proteins and induces apoptosis via the proteasome, mitochondria and caspase cascades in SCC25 cells.

Citrus platymamma inhibits the expression of pro-inflammatory cytokines, inducible nitric oxide synthase, and cyclooxygenase-2 in RAW264.7 macrophage (RAW264.7 대식세포에서 Citrus platymamma의 iNOS, COX-2, 염증성 사이토카인 발현 억제 효과)

  • Kim, Sang Suk;Park, Kyung Jin;An, Hyun Joo;Choi, Young Hun
    • Food Science and Preservation
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    • v.23 no.7
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    • pp.1026-1032
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    • 2016
  • Citrus platymamma hort. ex Tanaka is widely used in traditional Korean medicine because of its medicinal benefits including an anti-inflammatory effect. This study aimed to evaluate changes in the flavonoid content and anti-inflammatory activities of C. platymamma during its harvest period. Fruit peel samples were obtained between September 2015 and February 2016. The results indicate that C. platymamma peel extract (CPE) was an effective inhibitor of lipopolysaccharide (LPS)-induced NO production in RAW264.7 cells. The inhibitory effects of CPE at $100{\mu}g/mL$ concentration included dose-dependent decreases in the expression of iNOS and COX-2 proteins. In addition, CPE decreased the expression of pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. The highest anti-inflammatory activity and flavonoid content were observed in CPE of C. platymamma peel harvested during the immature fruit period in September. Further, to assess the suitability of CPE for cosmetic use, we performed MTT assays using HaCaT keratinocytes and observed that CPE did not exhibit any cytotoxicity. To test the potential application of CPE as a cosmetic material, we also performed primary skin irritation tests on normal skin of 30 volunteers and no adverse reactions were observed. The results of this study indicate that CPE may be considered as an anti-inflammatory candidate for inclusion in cosmetic materials.

Anti-inflammatory Effects by Arctium lappa L. Root Extracts through the Regulation of ICAM-1 and Nitric Oxide (우엉뿌리추출물이 ICAM-1과 NO조절에 미치는 항염증효과)

  • Kim, Ye-Jin;Kang, Se-Chan;NamKoong, Seung;Choung, Myoung-Gun;Sohn, Eun-Hwa
    • Korean Journal of Plant Resources
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    • v.25 no.1
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    • pp.1-6
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    • 2012
  • Atopic dermatitis is a chronic, allergic inflammatory skin disease that is accompanied by pruritic chronic eczema and markedly increased levels of inflammatory cells in endothelial cells. Arctium lappa L. is a popular edible vegetable cultivated in Asia. This study examined the effect of butanol extracts of A. lappa (ALBE) on the expression of adhesion molecule, ICAM-1 and the production of NO-iNOS induced by TNF-alpha in A549 endothelial cells. We also studied the effects of ALBE on the proliferation of keratinocyte. We observed significant inhibition of NO-iNOS production in dose-dependant manners and ALBE at $100\;{\mu}g$/mL suppressed the expression of ICAM-1 in TNF-${\alpha}$-induced A549 cells. In addition, the treatment of ALBE for 48 hrs increased the proliferation of HaCaT keratinocytes. These results support that ALBE has an anti-inflammatory effects for the treatment of atopic dermatitis.

Evaluation of Antioxidant, Tyrosinase and Collagenase Inhibitory of Grateloupia elliptica Extracts after Aureobasidium pullulans Fermentation (흑효모를 이용한 참도박 발효 추출물의 항산화 효과와 티로시나제 및 콜라게나제 저해효과)

  • Vu, Van Vinh;Lee, Kyung Eun;Kang, Sang Gu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.1
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    • pp.1-9
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    • 2020
  • In this experiment, the Grateloupia elliptica (G. elliptica) was fermented using the fungus Aureobasidium pullulans (A. pullulans) and its extract was obtained from hot water and 70% ethanol solution. The extract was studied for their biological activities such as antioxidant effect, Collagenase and tyrosinase inhibition in comparison to the nonfermented exatract in same solvents. Antioxidative activity test using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) showed that ethanol extract had higher antioxidant activity than water extract. Among all of the samples, the antioxidant activity of G. elliptica fermented ethanol extracts (GEFEE) was highest. Tyrosinase inhibitory activity of GEFEE was highest with 9.8% at the 1,000 ㎍/mL concentration. No inhibition of collagenase from G. elliptica water extract (GEWE) and G. elliptica fermented water extract (GEFWE) was observed, but G. elliptica ethanol extracts (GEEE) and GEFEE showed increased collagenase inhibition activity with increasing concentrations of them. Collagenase inhibitory activity of GEFEE was highest with 50.3% at the 1,000 ㎍/mL concentration. MTS cell proliferation assay was conducted with the GEWE, GEEE, GEFWE, GEFEE and cell viability was over 90% at the 10 ㎍/mL ~ 1000 ㎍/mL concentrations for all of the samples, which suggested that the extracts were noncytotoxic. In conclusion, fermented extracts of G. elliptica could be developed to bioactive functional material for cosmetics with antioxidant and wrinkle improvement effects.

Evaluation of the Physiological Activity and Identification of the Active Ingredients of Crab Apple (Malus prunifolia Borkh.) Extracts (꽃사과(Malus prunifolia Borkh.) 추출물의 생리활성 평가 및 활성 성분의 규명)

  • Shin, Hyun Young;Kim, Hoon;Jeong, Eun-Jin;Kim, Hyun-Gyeong;Lee, Kyung-Haeng;Bae, Yun-Jung;Kim, Woo Jung;Lee, Sanghyun;Yu, Kwang-Won
    • The Korean Journal of Food And Nutrition
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    • v.34 no.5
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    • pp.477-486
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    • 2021
  • To utilize Malus pruniforia Borkh. as a functional material, cold-water (CW), hot-water (HW), and 70% ethanol (EtOH) extracts were prepared, and their antioxidant and anti-inflammatory activities were compared. The antioxidant activity of the HW extract evaluated by ABTS and DPPH radical scavenging and FRAP activity was significantly effective. The total polyphenol content of the HW extract was also higher by 15.5±0.7 mg GAE/g extract compared to other extracts. The EtOH extract showed significantly decreased TNF-α (39.8%), IL-6 (65.5%), and NO (34.9%) levels in RAW 264.7 cells compared to the LPS-induced control group. The levels of IL-6 (21.1%) and IL-8 (19.3%) were significantly decreased by treatment of EtOH extract in HaCaT keratinocytes induced with TNF-α and IFN-γ. The UHPLC-MS results indicated that the EtOH extract might have chlorogenic acid and phlorizin as the major compounds. This was validated using HPLC-DAD, which showed that the EtOH extract had higher levels of chlorogenic acid and phlorizin (1,185±58 and 470±10 ㎍/g extract, respectively). In conclusion, the present study suggested that the anti-inflammatory activity of the EtOH extract was more effective than the CW and HW extracts, and chlorogenic acid and phlorizin could be used as indicator compounds and functional substances.