• Title/Summary/Keyword: anti-oxidant assay

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Protective effect of lycopene against cytokine-induced β-cell apoptosis in INS-1 cells (라이코펜이 사이토카인에 의해 유도된 베타세포 사멸에 미치는 효과 및 기전 연구)

  • Kim, Kyong;Jang, Se-Eun;Bae, Gong Deuk;Jun, Hee-Sook;Oh, Yoon Sin
    • Journal of Nutrition and Health
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    • v.51 no.6
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    • pp.498-506
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    • 2018
  • Purpose: Lycopene, a carotenoid with anti-oxidant properties, occurs naturally in tomatoes and pink grapefruit. Although the beneficial effects of lycopene on various disorders have been established, little attention has been paid to the possible anti-diabetic effects of lycopene focusing on ${\beta}$-cells. Therefore, this study investigated the potential of lycopene to protect ${\beta}$-cells against apoptosis induced by a cytokine mixture. Methods: For toxicity experiments, the cells were treated with 0.1 ~ 10 nM of lycopene, and the cell viability in INS-1 cells (a rat ${\beta}$-cell line) was measured using a MTT assay. To induce cytokine toxicity, the cells were treated with a cytokine mixture (20 ng/mL of $TNF{\alpha}$ + 20 ng/mL of IL-$1{\beta}$) for 24 h, and the effects of lycopene (0.1 nM) on the cytokine toxicity were measured using the MTT assay. The expression levels of the apoptotic proteins were analyzed by Western blotting, and the level of intracellular reactive oxidative stress (ROS) was monitored using a DCFDA fluorescent probe. The intracellular ATP levels were determined using a luminescence kit, and mRNA expression of the genes coding for anti-oxidative stress response and mitochondrial function were analyzed by quantitative reverse-transcriptase PCR. Results: Exposure of INS-1 cells to 0.1 nM of lycopene increased the cell viability significantly, and protected the cells from cytokine-induced death. Lycopene upregulated the mRNA and protein expression of B-cell lymphoma-2 (Bcl-2) and reduced the expression of the Bcl-2 associated X (Bax) protein. Lycopene inhibited apoptotic signaling via a reduction of the ROS, and this effect correlated with the upregulation of anti-oxidative stress response genes, such as GCLC, NQO1, and HO-1. Lycopene increased the mRNA expression of mitochondrial function-related genes and increased the cellular ATP level. Conclusion: These results suggest that lycopene reduces the level of oxidative stress and improves the mitochondrial function, contributing to the prevention of cytokine-induced ${\beta}$-cell apoptosis. Therefore, lycopene could potentially serve as a preventive and therapeutic agent for the treatment of type 2 diabetes.

Anti-oxidant Function and Inhibitory Effects of the Expression of MITF, TRP-1, TRP-2 and Tyrosinase of Sesamum indicum L. in B16F10 Melanoma Cells (참깨 추출물의 항산화 활성 및 melanoma cell (B16F10)에서 MITF, TRP-1, TRP-2, tyrosinase 의 발현 저해)

  • Yoo, Dan-Hee;Joo, Da-Hye;Lee, Jin-Young
    • Journal of Life Science
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    • v.27 no.3
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    • pp.318-324
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    • 2017
  • This study was performed to improve the antioxidant and skin-whitening activities of 70% ethanol extract from Sesamum indicum L. (SIL). The electron-donating ability of the SIL extract was 71.7% at a concentration of $1,000{\mu}g/ml$. The whitening effects that was measured by tyrosinase inhibition assay. As a result, SIL extract was shown 42% at $1,000{\mu}g/ml$ concentration. The cell toxicity on B16F10 melanoma cells of SIL of 70% ethanol extract showed 84.3% at $1,000{\mu}g/ml$ concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase relate protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2) and Tyrosinase protein and mRNA expression inhibitory effect of SIL extract were measured by western blot and reverse transcription- polymerase chain reaction (PCR) at 50, 250, $500{\mu}g/ml$ concentration. Consequently, the MITF, TRP-1, TRP-2, Tyrosinase protein expression inhibitory effect of SIL extract was decreased by 68.3%, 39.2%, 89.7%, 22.3%, respectively, at $500{\mu}g/ml$ concentration. Moreover, MITF, TRP-1, TRP-2, Tyrosinase mRNA expression inhibitory effect by reverse-transcription-PCR of SIL extract was decreased by 81.8%, 66.5%, 84.2%, 68.1%, respectively, at $500{\mu}g/ml$ concentration. Therefore, we excellently identified the antioxidant activities and whitening effect of SIL extract, and this finding suggested that SIL extract has great potential as a cosmetic ingredients.

Antioxidant Activity of Methanol Extracts from $Lactuca$ $indica$ (왕고들빼기 메탄올 추출물의 항산화성)

  • Kim, Ju-Nam;Kim, Ja-Min;Lee, Kyung-Soo
    • Food Science and Preservation
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    • v.19 no.2
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    • pp.294-300
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    • 2012
  • The anti-oxidant properties of $Lactuca$ $indica$ were determined using in-vitro assay systems. The vitamin C contents of the leaf and root extracts were 24.14 and 0.38 mg/100 g, respectively. The total polyphenol contents of the leaf and root extracts were 42.8 and 7.66 mg/g, and their flavonoid contents were 23.09 and 0.77 mg/g. The leaf extract showed higher DPPH and ABTS radical scavenging ability than the root extract at all the extract concentrations. Especially, the ABTS radical scavenging ability of the leaf extract was 92.3% at a concentration of 5 mg/mL. The reducing power was increased with the increase in the concentration of extracts, and the leaf extract had a higher reducing power than the root. The $Fe^{2+}$-chelating ability of the leaf and root were 97.2% and 34.3% at 14 mg/mL, respectively. The $IC_{50}$ values of the leaf for DPPH, its ABTS radical scavenging ability, and its $Fe^{2+}$-chelating ability were 0.19, 2.7, and 6.27 mg/mL, respectively, and the leaf extract showed lower $IC_{50}$ values than root extract. These results show that the $L.$ $indica$ leaf extract contained high amounts of anti-oxidative compounds and had higher anti-oxidant activity levels than the root extract. It is suggested that Lactuca indica is very high in availability as a functional food and in its materials.

Effects of Neutral Dammarane Saponin from Panax ginseng on the in vitro Function of Polymorphonuclear Leukocytes (인삼의 중성 Dammarane계 사포닌의 다형핵 백혈구 기능에 미치는 영향)

  • Bridges Raymond B.;Park Ki Hyun;Han Byung Hoon;Han Yong Nam;Chung Soo Il
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.115-121
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    • 1988
  • Although Saponin A from Panax ginseng has previously been shown to inhibit carageenin induced edema. a paucity of information exists on the effects of components from ginseng on the cellular inflammatory response. specifically polymorphonuclear leukocyte (PMNL) function. The purpose of this study was 10 determine the effects of isolated neutral dammarane saponins from ginseng (i.e..glycosidic derivatives of 20(S)-protopanaxadiol [ginsenoside $Rb_1,\;Rb_2$ and Rc] and 20(S)-protopanaxatriol [ginsenosides Re and $Rg_1$]) on in vivo PMNL function and to compare their effects with those produced by a steroidal anti-inflammatory agent (dexamethasone) and commercially available saponin. Dexamethasone. the ginsenosides and saponin were all shown to he potent inhibitors of PMNL chemotaxis using the $^{51}Cr$ assay with $5{\times}10^{-8}M$ f-met-leu-phe [FMLP] as the chemoattractant. Inhibition or PMNL chemotaxis by dexamethasone. the ginsenosides and saponin were all shown to be both time-and dose-dependent and these agents did not affect cellular viability at the concentrations tested Saponin and the ginsenosides were more potent inhibitors of chemotaxis than was dexamethasone. while oxidant generation (as measured by the luminol-enhaneed chemil-uminescence of PMNL using FMNL $[10^{-6}]$ as the stimulus) was inhibited by dexamethasone. the ginsenosides $(Rb_1\;Rb_2\;Rc\;Re\;and\;Rg_1)$ and saponin at a concentration of 1 ${\mu}M$ had no significant effect on PMNL chemiluminescence. Thus. the neutral dammarane saponins are potentially important modulators or PMNL function and their inhibitory effects may he differentiated from those of the Steroidal anti-inflammatory agents.

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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.

Study on the Physiological Activity of Chrysanthemum indicum Flower Extracts Using the Hydrothermal Water Extraction Method (감국꽃 열수 추출물의 생리활성에 관한 연구)

  • Jisung Ha;Yuri Kang;Woonjung Kim
    • Industry Promotion Research
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    • v.9 no.3
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    • pp.77-85
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    • 2024
  • In this study, the hydrothermal extract of Chrysanthemum indicum Flowers are analyzed for the identification of its functional material value. Various aromatic compounds were detected as a result of GC-MS analysis, and antioxidant analysis confirmed the high antioxidant activity of the extract. High antibacterial activity was confirmed in the TIME KILL TEST using the 70% ethanol extract, which showed the highest activity, and the inhibition of intracellular melanogenesis was confirmed to be 115.77 ± 6.67%, 112.42 ± 1.64%, and 67.12 ± 3.89% compared to the α-MSH treatment group at concentrations of 1%, 1.2%, and 1.5%, respectively, when the extract was treated with 1%, 1.2%, and 1.5%. Collagenage activity inhibition assay showed collagenase inhibition of 56.57 ± 2.28%, 58.64 ± 2.99 and 29.53 ± 1.03% when treated at concentrations of 1.5%, 3% and 5%. The cytotoxicity test of the extract showed no cytotoxicity at concentrations of 1.5%, 3% and 5%. Therefore, this study predicts that the hydrothermal extract of Chrysanthemum indicum Flowers will be useful as a functional cosmetic material.

Anti-aging and Anti-diabetes Effects of Aconitum pesudo-laeve var. erectum Extracts (진범(Aconitum pesudo-laeve var. erectum) 추출물의 항노화 및 항당뇨 효과)

  • Kim, Jeung-Hoan;Lee, Soo-Yeon;Kwon, O-Jun;Park, Joo-Hoon;Lee, Jin-Young
    • Journal of Life Science
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    • v.23 no.5
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    • pp.616-621
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    • 2013
  • Aconitum pesudo-laeve var erectum has been known to possess anti-inflammatory activity and modulate the intestinal immune system. In addition, it has traditionally been used for the treatment of water retention in the body. In this study, the anti-aging and anti-diabetes effects of water and ethanol extracts from Aconitum pesudo-laeve var. erectum were investigated. The activities of each extract were measured by antioxidant tests such as DPPH and ABTS radical scavenging activity, antioxidant protection factor (PF), TBARs content, and ${\alpha}$-amylase and ${\alpha}$-glucosidase inhibition activity assay. DPPH radical scavenging activity was found in over 50% of water and ethanol extracts at $100{\mu}g/ml$, $50{\mu}g/ml$, respectively. The ABTS radical scavenging activity of ethanol extract was $99.8{\pm}0.1$% at $1,000{\mu}g/ml$ in water, which was highest among the ethanol extract concentrations. PFs measured with ${\beta}$-carotene-linoleate model systems were in the order of ethanol (1.49 PF at $1,000{\mu}g/ml$) > ethanol (1.40 PF at $500{\mu}g/ml$) > water (1.33 PF at $1,000{\mu}g/ml$) > water (1.27 PF at $500{\mu}g/ml$). TBARs content in ethanol extracts ($1,000{\mu}g/ml$) was $0.16{\pm}0.03{\mu}M$, which was lower than that of water extracts and other ethanol extract concentrations. The extracts also showed over 90% of ${\alpha}$-amylase inhibition and over 60% of ${\alpha}$-glucosidase inhibition ratio in water ($1,000{\mu}g/ml$) and ethanol extracts (100~$1,000{\mu}g/ml$). These results suggest that Aconitum pesudo-laeve var. erectum extracts could be used as a cosmetic source and preventive agent for aging and diabetes.

Anti-proliferation, Cell Cycle Arrest, and Apoptosis Induced by Natural Liquiritigenin from Licorice Root in Oral Squamous Cell Carcinoma Cells (구강편평세포암종 세포에서 감초 유래 Liquiritigenin의 항증식, 세포주기 정지 및 세포사멸 유도)

  • Kwak, Ah-Won;Yoon, Goo;Chae, Jung-Il;Shim, Jung-Hyun
    • Journal of Life Science
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    • v.29 no.3
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    • pp.295-302
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    • 2019
  • Liquiritigenin (LG) is a chiral flavonoid isolated from the roots of licorice. It exhibits multiple biological activities including anti-oxidant, anti-cancer, and anti-inflammatory effects. In particular though, the anti-cancer activity of LG in oral squamous cell carcinoma has yet to be elucidated, and LG-induced apoptosis in oral squamous cell carcinoma remains poorly understood. In the present study, we tested the role of LG in inducing apoptosis in oral squamous cell carcinoma cells. LG treatment of HN22 cells resulted in a dose-dependent inhibition of cell viability as detected by a 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay. The induction of apoptosis in terms of Annexin V/7-Aminoactinomycin D staining, sub-G1 population, and multi-caspase activity were assessed with a $Muse^{TM}$ Cell Analyzer. Flow cytometric analysis revealed that LG treatment resulted in G2/M arrest in cell cycle progression and downregulation of cyclin B1 and CDC2 expression in a concentration-dependent manner. It also resulted in significant upregulation of p27. In addition, LG was seen to trigger the generation of reactive oxygen species and induce CCAAT/enhancer-binding protein homologous protein and 78-kDa glucose-regulated protein in concentration-dependent upregulation. The LG treatment of HN22 cells led to a loss of mitochondrial membrane potential (${\Delta}{\Psi}m$); it also reduced the levels of anti-apoptotic protein and increased the expression of apoptotic protease activating factor-1, cleaved poly (ADP-ribose)polymerase and Bax. Overall, our results indicate that the pro-apoptotic effects of LG in HN22 cells depend on the activation of both intrinsic and extrinsic signaling pathways. Thus, our results suggest that LG constitutes a natural compound with a potential role as an anti-tumor agent in oral squamous cell carcinoma.

Anti-inflammatory Efficacy and Liver Protective Activity of Pine Pollen according to Probe Sonicator Ultrasonic Disintegration Extraction Method (송화분의 초음파 파쇄 추출 방법에 따른 항염증 효능 및 간 보호 활성)

  • Kim, Ok Ju;Woo, Young Min;Jo, Eun Sol;Jo, Min Young;Li, Chun-Ri;Lee, Young-Ho;Ahn, Mee Young;Lee, Sang-Hyeon;Ha, Jong Myung;Kim, Andre
    • Applied Chemistry for Engineering
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    • v.30 no.5
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    • pp.569-579
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    • 2019
  • In this study, the effect anti-oxidant, anti-inflammatory, and liver protective activity was investigated via quick ultrasonic disintegration of pine pollen using a probe sonicator (PS) followed by the extraction with water, 70% ethanol, and 100% ethanol. The anti-inflammatory effect was studied by measuring the production of nitric oxide (NO) and cytokine in RAW264.7 cells induced with lipopolysaccharides (LPS). The cell toxicity was also checked with an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the experiment was conducted using non-toxic $100{\mu}g/mL$. The NO inhibition rate was highest in the 70% ethanol PS group at $85.99{\pm}0.12%$. Also an excellent efficiency was obtained from the results of interlukin-1 beta ($IL-1{\beta}$) and tumor necrosis factor alpha ($TNF-{\alpha}$), which is related to inflammation-related cytokine, with the respective inhibition rates of 63 and 22%. To examine liver protective activity, HepG2 cells were treated with Taclin, and the generation of glutamic oxaloacetic transaminase (GOT) and lactate dehydrogenase (LDH) was measured in the culture solution. From GOT and LDH generation results, the inhibition rates in the 70% ethanol PS group were 28% and 13%, respectively, which was higher compared to that of using negative control group. Our results suggest that pine pollen extracted in 70% ethanol using PS may be used to develop food products that have anti-aging, anti-inflammatory, and liver protective effects.

Anti-tumor Effect of a Combination of Hongyoung Ethanol Extract and Cisplatinin YD-10B Oral Cancer Cells (YD-10B 구강암세포에서 홍영 에탄올 추출물과 시스플라틴 병용에 의한 항암 효과)

  • Eun-Jung Kim;Sung-Hee Hwang;Sangwook Park
    • Journal of Life Science
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    • v.33 no.6
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    • pp.498-505
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    • 2023
  • Solanum tuberosum Linnaeus cv Hongyoung, which represents red potato, was developed in Korea. Hongyoung is known to have anti-oxidant, anti-inflammatory, anti-viral, and anti-tumor properties, but no research has been conducted on the growth inhibition and apoptosis effects of hongyoung in YD-10B oral cancer cells. In this study, the combined treatment of hongyoung ethanol extract (HEE) and cisplatin were examined to determine its ability to inhibit cancer cell growth, induce apoptosis, and inhibit matrix metalloproteinases (MMP)-2 and MMP-9 cancer metastasis. The cell viability was investigated using a 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H- tetrazolium monosodium salt (MTS) assay, and the ability to induce apoptosis was analyzed using an FACS analyzer. The mRNA expression and protein activity of MMP-2 and MMP-9 were measured via RT-PCR and zymography. The YD-10B oral cancer cells showed an increase in growth inhibition as the concentration of HEE increased. The combination of 200 µM cisplatin and 500 ㎍/ml HEE reduced the growth of the YD-10B oral cancer cells by more than 50% compared to cisplatin alone. When phorbol 12-myristate 13-acetate (PMA)-treated YD-10B oral cancer cells were co-treated with 200 µM cisplatin and 500 ㎍/ml HEE, both the mRNA expression and protein activity of the MMP-2 and MMP-9 decreased. In addition, the percentage of the sub-G1 phase, which indicates apoptosis ability, more than doubled when treated in combination with 200 µM cisplatin and 500 ㎍/ml HEE than when cisplatin alone was used. The results of this study therefore suggest the possibility of using a combination of HEE and cisplatin in the development of effective drugs to treat oral cancer.