• Title/Summary/Keyword: HaCaT cell

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Investigation of the effect of Hibiscus sabdariffa L. extracts on tight-junction related genes in human keratinocyte HaCaT cells (히비스커스 추출물이 인간 각질 형성 세포의 밀착 연접 관련 유전자 발현에 미치는 영향 연구)

  • Jung, Haesoo;Kim, Eunmi;Han, Hyosang;Kim, Keekwang
    • The Korea Journal of Herbology
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    • v.36 no.5
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    • pp.59-67
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    • 2021
  • Objectives : Hibiscus (Hibiscus sabdariffa L.) is rich in antioxidants such as flavonoids and anthocyanins and is known to have anti-inflammatory activity and anti-aging function of the skin, but there is no study on its effect on the skin barrier. This study aim to investigate the positive effect on the skin barrier by confirming the effect of water extracts of Hibiscus sabdariffa L. (WEHS) on the tight junction-related gene expression. Methods : The antioxidant efficacy of WEHS was investigated through ABTS and DPPH assays, and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium was performed to examine the effect on cell viability. quantitative Reverse transcription polymerase chain reaction and immunoblot analysis were performed to confirm the effect of WEHS on the expression of tight junction-related genes, and immunofluorescence microscopy was used to confirm the movement of Claudin 1 protein into tight junctions. Results : WEHS showed strong antioxidant activity and induced an increase in both mRNA and protein expression levels of Claudin 1 among tight junction-related genes. The strong localization of Claudine 1 protein increased by WEHS to the tight junction was confirmed by immunofluorescence microscopy. Conclusions : Hibiscus was confirmed through this study to show antioxidant activity and the function of promoting the expression of the tight junction Claudin 1 gene, suggesting that Hibiscus can be used as a material for the prevention and treatment of skin moisturizing and atopy, which have an important influence on tight junction.

Therapeutic Effects of Amnion-Conjugated Chitosan-Alginate Membranes on Diabetic Wounds in an Induced Diabetic Swine Model: An In Vitro and In Vivo Study

  • Jeong, Woonhyeok;Hong, Jamin;Jung, Minho;Jang, Mijin;An, Sanghyun;Jo, Taehee;Kwon, Sunyoung;Son, Daegu
    • Archives of Plastic Surgery
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    • v.49 no.2
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    • pp.258-265
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    • 2022
  • Background Chitosan (CS) is a well-known antimicrobial dressing material. Moreover, widely used amniotic membranes contain growth factors beneficial for wound healing. Herein, we created a novel amnion-conjugated CS-alginate membrane dressing and tested its wound healing potency in a diabetic swine model. Methods The bovine amniotic powder growth factor contents were evaluated by protein assay, and the powder's wound healing effects were assessed in vitro by HaCaT cell scratch closure. In vivo, two minipigs developed streptozotocin-induced diabetes. Serial serum glucose measurements and intravenous glucose tolerance tests were performed to confirm their diabetic status. Twelve square-shaped wounds created on each pig's back were randomly divided into control (n = 4), CS (n = 4), and amnion-CS (AC; n = 4) groups and treated accordingly with different dressings. Wound healing in each group was assessed by measuring wound contraction over time, capturing wound perfusion with indocyanine green (ICG) angiography, and histologically analyzing inflammatory markers. Results Amniotic powder elution promoted HaCaT cell migration in the scratch wound model, suggesting its beneficial in vitro wound healing effects. In vivo, the CS and AC groups showed earlier wound contraction initiation and reepithelialization and earlier wound perfusion improvement by ICG angiography than the control group. Additionally, the wound size of the AC group at week 3 was significantly smaller than those in the control group. There was no significant difference in the numbers of acute and chronic inflammatory cells between the groups. Conclusion The amnion-conjugated CS-alginate membrane, as well as CS dressing alone, could be a favorable dressing option for diabetic wounds.

Effect of Fermented Soybean on the Proliferation and Growth in HaCaT and Fibroblast Cell (대두 발효물이 인간 유래 피부세포의 증식 및 성장에 미치는 영향)

  • Kim, Eun-Joo;Han, Myung-Ryun;Lee, So-Young;Kim, Ae-Jung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.2
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    • pp.326-335
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    • 2021
  • This study was undertaken to determine the effect of fermented soybean extract and its fractions on skin cell proliferation and growth. The extract was procured by the pepsin and Lactobacillus rhamnosus fermentation of soybean. LC-MS analysis was performed subsequent to soybean fermentation, and cell viability was measured by the WST-1 assay. Cell proliferation was observed to increase after exposing cells to the fermented soybean extract and its fractions at all concentrations tested (0~2,000 ㎍/mL). In particular, compared to the normal control group and 120 % proliferation of the EGF (epidermal growth factor) positive control group, 160~180 % cell proliferation was achieved at 800 ㎍/ml, indicating the excellent potential as an application material for skin aging inhibition and skin cell regeneration. In addition, we also examined the effects of fermented soybean extract and its fractions on wound healing ability, in HaCaT cells and fibroblasts. Our results indicate excellent cell migration abilities after treatment with fermented soybean extract and its fractions, as compared to the control treatment. Similar cell migration abilities were observed in the positive control group (EGF). Taken together, our results indicate that fermented soybean extract and its fractions (F4 and F5) exert amelioratory effects as a natural material for skin.

Protective Effects of Trifolium pratense L. Extract against H2O2-induced Oxidative Stress in HaCaT Keratinocytes (인간 피부각질세포에서 Hydrogen peroxide로 유도된 산화적 스트레스에 대한 붉은 토끼풀 추출물의 세포 보호 효과)

  • Mi Song Shin;You Kyeong Lee;Seo Young Choi;Ji Sun Hwang;Parkyong Song;Hyeon Cheal Park;Keun Ki Kim;Hong-Joo Son;Yu-Jin Kim;Kwang Min Lee
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.2
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    • pp.223-232
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    • 2023
  • Oxidative stress plays a significant role in the pathogenesis of various skin conditions, resulting in cellular and tissue damage that can contribute to the development of skin tone unevenness, roughness and wrinkles. In this study, we found that Trifolium pratense L. extract (TE) attenuated oxidative-induced damage in HaCaT cells and elucidated the underlying molecular mechanism. Our finding demonstrated that TE effectively protected HaCaT cells against H2O2-induced cell death by inhibiting caspase-3 activation, downregulating Bax and upregulating Bcl-2, and attenuating the activation of three mitogen-activated protein kinases (MAPKs). Our results suggest that TE has remarkable cytoprotective properties against oxidative damage in HaCaT cells and could serve as a complementary or alternative approach to prevent and treat skin damage.

Effects of Bambusae Caulis in Taeniam Extract on the UVB-induced Cell Death, Oxidative Stress and Matrix Metalloproteinase 1 Expression in Keratinocytes (각질세포에서 자외선B가 유도한 세포 사멸, 산화적 스트레스 및 matrix metalloproteinase 1 발현에 대한 죽여추출물의 영향)

  • Seok, Jin Kyung;Kwak, Jun Yup;Seo, Hyeong Ho;Suh, Hwa Jin;Boo, Yong Chool
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.1
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    • pp.9-20
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    • 2015
  • Ultraviolet radiation (UV) is a major cause of skin photoaging, and effective UV protecting agents are needed for the skin health and beauty. This study was undertaken to examine the effects of Bambusae caulis in Taeniam extract (BCTE) on UVB-induced cell death, oxidative stress and matrix metalloproteinase 1 (MMP1) expression in cell-based assays. HaCaT human keratinocytes were exposed to UVB in the presence of BCTE at different concentrations and resulting changes in cell viability and biochemical events were determined. The results showed that BCTE enhanced the viabilities of UVB-exposed cells, and attenuated apoptotic events such as cleavage of procaspase 3 to its active form, and the increase of Bax to Bcl-2 ratios. BCTE also attenuated the reactive oxygen generation and lipid peroxidation in cells exposed to UVB. Additionally, it attenuated the expression of matrix metalloproteinase 1 and the phosphorylation of c-Jun N-terminal kinase stimulated by UVB. Conclusively, the present study demonstrated that BCTE pro tected skin cells from the UVB-induced cell death, oxidative stress and MMP1 expression, suggesting its potential use as a cosmetic ingredient mitigating some features of the skin photoaging.

Anti-inflammatory and Antioxidant Effects of Water Extracts of Sasangja-tang(SSJ) and Gami-sasangja-tang(GSJ) (사상자탕과 가미사상자탕의 항염증 및 항산화 효과 비교 연구)

  • Choi, Jung-Eun;Park, Bo-Kyung;Jin, Mirim
    • Journal of Haehwa Medicine
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    • v.23 no.2
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    • pp.5-13
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    • 2015
  • Objectives : The anti-inflammatory and antioxidant effects of water extracts of Sasangja-tang(SSJ) and Gami-sasangja-tang(GSJ) were investigated. The effects of SSJ and GSJ were compared. Methods : We performed cell viability assay in HaCaT cells and RAW 264.7 cells using 3-(4,5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide(MTT) assay. We measured chemokines(regulated on activation normal T-cell expression and secreted ; RANTES/CCL5, interferon-inducible protein; IP-10/CXCL10, macrophage-derived chemokine; MDC/CCL22) in HaCat cells, also we measured cytokines (tumor necrosis factor-${\alpha}$; TNF-${\alpha}$, interleukin-6; IL-6) and nitric oxide(NO) production in RAW 264.7 cells using enzyme-linked immunosorbent assay(ELISA) and NO assay. Western blot assay was used to evaluate the expression for inducible nitric oxide synthase(iNOS) in RAW 264.7 cells. Results : SSJ and GSJ did not affect the cell viability at the concentrations treated ($0-800{\mu}g/ml$). As a result of SSJ and GSJ treatment in HaCat cells stimulated by TNF-${\alpha}$(10 ng/ml) and interferon(IFN)-${\gamma}$(10 ng/ml), the production of RANTES and IP-10 was inhibited significantly. However there was no significant difference in the secretion of MDC. And in RAW 264. 7 cells stimulated by lipopolysaccharide(LPS, $1{\mu}g/ml$), SSJ and GSJ treatment significantly inhibited the secretion of TNF-${\alpha}$ and IL-6 and the production of NO. The expression of iNOS was also decresed by SSJ and GSJ treatment in RAW 264. 7 cells. Compared with SSJ, GSJ was superior to SSJ in inhibition of RANTES, IP-10, TNF-${\alpha}$, IL-6 and NO production at the concentration of $200{\mu}g/ml$. Conclusion : Both SSJ and GSJ have anti-inflammatory and antioxidant effects. And GSJ has better effects than SSJ.

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Anti-Microbial and Anti-Wrinkle Effect of Kaempferol and Kaempferol Rhamnosides isolated from Hibiscus cannabinus L. (양마에서 분리한 Kaempferol과 그 배당체의 항균 및 주름억제 효과)

  • Lee, Keun Ha;Kong, Hae Jin;Cho, Young Long;Joo, Chul Gue;Kwon, Sun Sang;Hwang, Jae Sung;Park, Chung
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.454-460
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    • 2012
  • In this study, kaempferol and its rhamnosides (${\alpha}$-rhamnoisorobin, afzelin, kaempferitrin) were isolated electively by bioconversion technology from Hibiscus cannabinus L. leaves to evaluate the anti-wrinkles effects and anti-microbial effects. In order to evaluate anti-wrinkles activity, reduction of expression matrix metalloproteinase-1 (MMP-1) protein and proliferation/ pro-collagen production were investigated. Kaempferol and ${\alpha}$-rhamnoisorobin showed inhibition activity of MMP-1 generated to compared to positive control. In HaCaT cell proliferation assay, kaempferol and ${\alpha}$-rhamnoisorobin significantly promoted cell proliferation in a concentration-dependent manner. In addition, procollagen synthesis assays (by HDF-N cell) showd that TGF-${\beta}$ induced procollagen production and also, all four kinds of experimental significantly promoted procollagen synthesis in a concentration-dependent manner. Kaempferol and ${\alpha}$-rhamnoisorobin exhibited strong antimicrobial activities on five of microbes, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger.

Enhancement of Tissue Type Plasminogen Activator (tPA) Production from Recombinant CHO Cells by Low Electromagnetic Fields

  • Lee, Seo-Ho;Lee, Hyun-Soo;Lee, Mi-Kyoung;Lee, Jin-Ha;Kim, Jong-Dai;Park, Young-Shik;Lee, Shin-Young;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.457-462
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    • 2002
  • Low Electromagnetic Field (EMF) intensity in the range of $1{\mu}T\;to\;10{\mu}T$(Tesla) was found to enhance the growth of CHO cells and the production of tPA in batch and perfusion cultivations. At $1{\mu}T\;intensity,\;1.3{\times}10^7$ viable cells/ml of maximum cell density and 80 mg/l of maximum tPA production were obtained in batch cultivation, compared to $2.8{\times}10^6$ viable cells/ml and 59 mg tPA/1 in unexposed case (control). A similar trend was observed in the perfusion process, where it was possible to obtain $1.2{\times}10^7$ viable cells/ml of maximum cell density and 81 mg tPA/l of maximum tPA production by more than 80 days of cultivation. However, there was not much difference between $1{\mu}T\;and\;10{\mu}T$ in perfusion cultivation, possibly due to better environmental growth conditions being maintained by continuous feeding of fresh medium into the reactor. On the contrary, both cell growth and tPA production were severely inhibited at higher than 1 mT intensity, showing no growth at 10 mT exposure. Specific growth rate was linearly correlated to specific tPA production rate at $1{\mu}T$EMF intensity, which represents a partially growth-related relationship. It was also found that a large amount of $Ca^2+$ was released at low EMF intensity, even though the cell growth was not much affected. Low EMF intensity significantly improved both cell growth and tPA production, and tPA production seemed to be more affected than the cell growth, possibly due to the changes of cell membrane characteristics. It can be concluded that the elaboration of EMF intensity less than $10{\mu}T$ could improve cell growth and tPA production, but mainly tPA secretion through batch or perfusion process in a bioreactor.

Enhancement of Cell Migration by Corticotropin-Releasing Hormone (CRH) in Human Gastric Cancer Cell Line, MKN-28 (Corticotropin-Releasing Hormone (CRH)에 의한 인간 위암 세포(MKN-28)의 Migration 증가)

  • Cheon, Soyoung;Cho, Daeho
    • IMMUNE NETWORK
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    • v.4 no.4
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    • pp.244-249
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    • 2004
  • Background: Corticotropin-Releasing Hormone (CRH), an important regulator of stress response, has a potent immunoregulatory effect with the ability to promote the growth of various cancer through CRH receptor type 1 under stress. Although the metastasized cancers through cell migration are more aggressive than the primary cancers, little is known about the effect of CRH on cell migration. Gastric cancer is prone to metastasize to other tissues and it is reported that gastric cancer is response to various stresses such as oxidative stress. Herein, we studied the relationship between CRH and gastric cancer cell migration. Methods: We used gastric cancer cell line, MKN-28 and tested the CRH receptor type 1 expression on MKN-28 by RT-PCR. To examine the change in the ability of migration by CRH in MKN-28, cells were incubated with CRH and then migration ability was measured using a cell migration assay. Results: We confirmed that CRH receptor type 1 was expressed in MKN-28 and HaCaT cells. The migration ability of MKN-28 cells was increased by CRH in a time-, dose- dependent manner. Conclusion: These data suggest that CRH increases migration ability in gastric cancer cell line and that CRH may be a critical regulator in the metastasis of gastric cancer cell.

Nootkatol prevents ultraviolet radiation-induced photoaging via ORAI1 and TRPV1 inhibition in melanocytes and keratinocytes

  • Woo, Joo Han;Nam, Da Yeong;Kim, Hyun Jong;Hong, Phan Thi Lam;Kim, Woo Kyung;Nam, Joo Hyun
    • 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.