• Title/Summary/Keyword: HaCat normal cell

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A Cytotoxic Compound from Moutan Cortex Radicis (목단피의 세포독성 물질)

  • Chu, Bo-Yeun;Jin, Wen-Yi;Choi, Su-La;Myung, Pyung-Keun;Seong, Yeon-Hee;Song, Kyung-Sik;Bae, Ki-Hwan
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.3
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    • pp.255-261
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    • 2004
  • The MeOH extracts from 21 species were tested for their cytotoxicity against SK-MEL-28 melanoma cell line and HaCat normal cell line in 5 g/ml by sulforrhodamine-B (SRB) method. Among them, the MeOH extract from Moutan Cortex Radicis showed the moderate activity with the growth rate of 74.3% in SK-MEL-28 cells and the high activity with the growth rate of 207.8% in HaCat cells. Activity-guided fractionation was performed and five compounds, paeonol (1), benzoylpaeoniflorin (2), benzoic acid (3), 2,5-dihydroxy-4-methoxy- acetophenone (4), paeonfliorin (5) were isolated from hexane and EtOAc fraction. The structures were established by physicochemical and spectrometric methods $(mp,\;UV,\;IR,\;^1H-NMR,\;^{13}C-NMR,\;spectram)$. All compounds were evaluated for their cytotoxicity, compound 4 showed the significant cytotoxic activity with $ED_50$ value of $5.92\;{\mu}g/ml$, but the other compounds no activity. These results suggest that compound 4 is a novel anticancer candidate against SK-MEL-28 melanoma cells.

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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Inhibitory Effect of Retinoic Acid on lipid Synthesis in Human Sebocyte (피지선세포에서 Retinoic Acid의 피지생성억제효과)

  • Mun Yeun Ja;Kim Youn Seok;Kwon Gang Joo;Rhee Hee Sub;Roh Seong Taek;Kim Yang Jin;Lee Jang Cheon;Woo Won Hong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1317-1321
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    • 2004
  • The differentiation of the sebaceous gland is remarkably species-specific and sebocytes may play crucial parts in the pathophysiologic processes and disorders of pilosebaceous unit SZ95 cell is an immortalized human sebaceous gland cell line that shows characteristics of normal human sebocytes, In this study, we investigated the effect of testosterone and the anti-androgenic effect of 13-cis-retinoic acid (13-cis-RA) on lipid synthesis in SZ95 cells. Cytoplasmic lipid droplets were shown by Oil-red staining. The majority of the SZ95 cells positively labeled with Oil-red dye, while HaCaT cells negatively labeled with Oil red dye. Total lipid level of SZ96 cells is higher 4 times than that of HaCat cells. Testosterone markedly increased 2 times lipid synthesis of SZ95 cells in compared with control. 13-cis-RA significantly inhibited lipid synthesis and cell proliferation in SZ95 cells. Combined treatment with testosterone and 13-cis-RA resulted in a lower total lipid levels than that with androgen alone. In conclusion, SZ95 cells well resembled the morphologic and functional characteritics of normal human sebocytes. This in vitro model could provide a valuable tool for the study of sebocytes with a key role in pathophysiology and differentiation of sebaceous glands.

Comparative Analysis on Antioxidant Activity in Various Human Skin Fibroblasts and Mesenchymal Stem Cells (사람의 피부 섬유아세포 및 중간엽 줄기세포에서 항산화 활성의 반응에 대한 비교 분석)

  • Kong, Ji-Weon;Park, Ryeok;Park, Joon-Woo;Lee, Joo-Yeong;Choi, Yeon-Joo;Moon, Sun-Ha;Kim, Hyeon-Ji;Jeon, Byeong-Gyun
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.2
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    • pp.394-406
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    • 2019
  • The cellular senescence may be due to damage by the reactive oxygen species (ROS). This study has compared the antioxidant activity in the human cell lines of various origins, including 10S and 50S-derived normal skin fibroblasts, and 10S bone marrow, dental tissue and adipose-derived adult stem cells. After being exposed to $H_2O_2$, half inhibitory concentration ($IC_{50}$) values by cytotoxicity assay was significantly (P<0.05) lower in 50S-derived skin fibroblasts, than in 10S-derived skin fibroblasts and various adult stem cell lines. The cell population doubling time (PDT) and the cell frequency with high senescence associated-${\beta}$-galactose activity were remarkably increased in 50S-derived fibroblasts exposed to 50 ppm $H_2O_2$ for 7 days, than those of 10S-derived fibroblasts and various adult stem cell lines. Further, the expression level of antioxidant-related genes, glutathione peroxidase (GPX) and catalase (CAT), was investigated in 10S and 50S-derived skin fibroblasts, and 10S-derived various adult stem cells by reverse transcription polymerase chain reaction (RT-PCR). The expression level of GPX was higher in most of cell lines, compared to CAT, and a significantly (P<0.05) higher expression level of GPX was observed in 10S-derived skin fibroblasts and adult stem cell lines, compared to 50S-derived skin fibroblasts. We concluded that old-aged skin fibroblasts seemed to be less resistant against ROS than young-aged skin fibroblasts and adult stem cells.

Protective Effects of Succinic Acid of Succiniter against Liver Toxicity (간 독성에 대한 보석 호박 호박산의 간 보호 효과)

  • Kim, Hong-Bi;Ha, Bae-Jin
    • Journal of Life Science
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    • v.27 no.8
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    • pp.896-901
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    • 2017
  • This study was performed to investigate the protective effects of succinic acid of Succiniter against carbon tetrachloride ($CCl_4$)-induced hepatotoxicity in rats. After an adaptation period of one week, Sprague-Dawley rats were administered succinic acid of Succiniter at 200 mg/kg every day for 21 days. Then $CCl_4$ (3.3 ml/kg) was intraperitoneally injected into rats of the other groups except the normal group, five hours after the last treatment of succinic acid of Succiniter on day 21. The succinic acid-treated group showed 93.20% and 88.76% of inhibitory effects in aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities, respectively, compared with the $CCl_4-treated$ group. The succinic acid-treated group showed inhibition of malonedialdehyde (MDA) by 85.17% compared with the $CCl_4-treated$ group. The succinic acid-treated group in liver homogenate promoted effects of 38.65% and 47.99% in superoxide dismutase (SOD) and catalase (CAT), respectively, compared with the $CCl_4-treated$ group. In conclusion, the AST and ALT activities of the succinic acid-treated group were both decreased compared with the $CCl_4-treated$ group. The MDA level of the succinic acid-treated group was decreased compared with the $CCl_4-treated$ group. However, the SOD and CAT levels of the succinic acid-treated group in liver homogenate were both increased compared with the $CCl_4-treated$ group. Also, histological examinations showed that the liver cell necrosis and centrilobular congestion aggregation induced by $CCl_4$ were clearly eliminated by treatment with succinic acid of Succiniter. These results suggest that succinic acid of Succiniter has a protective effect against liver damage and could be used in the development of the appropriate drug.

Protective Effect of Eriobotrya japonica Lindl. on Hepatotoxicity by Carbon Tetrachloride (비파식초의 사염화탄소에 의한 간독성 보호 효과)

  • Shin, Yu-Bin;Ha, Bae-Jin
    • Journal of Food Hygiene and Safety
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    • v.33 no.1
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    • pp.77-82
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    • 2018
  • This study was carried out to investigate the effect of natural Eriobotrya japonica Lindl. vinegar on the liver protective effect of animals exposed to carbon tetrachloride. Eriobotrya japonica Lindl. vinegar (200 mg/kg) was administered at the same time for 28 days, and hepatotoxicity was induced by intraperitoneal injection of carbon tetrachloride on the $29^{th}$ day. The aspartate aminotransferase and alanine aminotransferase levels were significantly decreased (p < 0.001) and the superoxide dismutase and catalase activities were significantly increased (p < 0.001) in the Eriobotrya japonica Lindl. vinegar group compared to the control group. Histopathological observations showed that the Eriobotrya japonica Lindl. vinegar showed hepatic cell structure similar to normal group, and these results showed that it had an effect of suppressing and protecting the damage of liver cell. Therefore, Eriobotrya japonical Lindl. vinegar is considered to be a healthy functional food of the liver.

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.