• Title/Summary/Keyword: MTT assay

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Induction of Apoptotic Cell Death by Red Pericarp Rice (Jakwangchalbyeo) Extracts

  • Chi, Hee-Youn;Lee, Chang-Ho;Kim, Kwang-Ho;Kim, Sun-Lim;Chung, Ill-Min
    • Food Science and Biotechnology
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    • v.15 no.4
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    • pp.534-542
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    • 2006
  • The effects of ethanol fractions of three different rice grain extracts, Jakwangchalbyeo, Hwasunchalbyeo, and Ilpumbyeo, on apoptotic cell death in the rat hepatoma H4IIE cell line were investigated using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cell viability assay. One hundred mg/mL Jakwangchalbyeo extract significantly reduced cell viability to 69.5, 57.2, and 46.1% within 24, 48, and 72 hr, respectively. Fluorescence-activated cell sorting (FACS) analyses were also performed to characterize the cell death pattern caused by treatment with the rice grain extracts. Apoptotic cell death was clearly observed with time after treatment with the Jakwangchalbyeo extract. In Western blotting analysis, degradation of the 116 kDa poly-ADP-ribose polymerase (PARP) molecule was observed with concomitant formation of an 89 kDa product 24, 48, and 72 hr after treating cells with the Jakwangchalbyeo extract. This indicates that an apoptotic process caused cell death in these cells. In conclusion, red-pericarp Jakwangchalbyeo extract induced apoptotic cell death in H4IIE cells to a larger extent than the other rice extracts.

An Experimental Study on the Effect of Angelica gigas Ethanol Extract on Hyaluronic Acid Synthesis (Angelica gigas 에탄올 추출물의 Hyaluronic acid 합성 효과에 대한 실험적 연구)

  • Park, Hye-Su;Ha, Hun-Yong;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.31 no.1
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    • pp.32-41
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    • 2018
  • Objectives : Hyaluronic acid(HA) is a mucopolysaccharide, occuring naturally in living organisms. It is one of the most hydrophilic molecules, so it has been known as being related to skin hydration and skin aging. The purpose of this study is to examine the effects of Angelica gigas(A. gigas) ethanol extract on hyaluronic acid synthesis. Methods : To determine cytotoxicity and hyaluronic acid synthase 2 gene expression, hyaluronic acid production in HaCaT cells, MTT assay and RT-PCR ELISA was used. Results : There were no cytotoxicity in $50{\mu}g/ml$ concentration A. gigas extract in MTT assay. Hyaluronic acid synthase 2(HAS2) gene expression was increased by all treated concentration A. gigas extract. Hyaluronic acid production was higher than control group in $50{\mu}g/ml$ & $100{\mu}g/ml$ concentration A. gigas extract. Conclusions : Hyaluronic acid production was increased by A. gigas extracts. Therefore, We suggest that A. gigas can make a contribution to the moisturizing effect on human skin.

The Inhibitory Effects of Houttuynia cordata $T_{HUNB}$ against Cadmium induced Cytotoxicity (II) (어성초의 카드뮴에 대한 독성억제효과 (II))

  • 이정호;유일수;이기남;정우영;한두석;백승화
    • YAKHAK HOEJI
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    • v.44 no.5
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    • pp.432-439
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    • 2000
  • This study was condo더ed to investigate the antitoxic component in aqueous extract of Houttuynia cordata $T_{HUNB}$. The results were as follows: Generally, detoxification effects by Houttuynia cordata $T_{HUNB}$ extract increased in proportion to the extract concentrations in rats. When 40 mBtg dosage of Houttuynia cordata $T_{HUNB}$ extract was administrated, Houttynia cordata $T_{HUNB}$ extract showed the highest antitoxic effects in metallothionein induction. After the extract treatment, body weights increased in proportion to the extract concentrations. However, after 3 weeks, the body weight decreased insignificantly. From the above results, Houttuynia cordate $T_{HUNB}$ extract increased metallothionein concentration and decreased the toxicity of cadmium in rats. In vitro the antitoxic activity of aqueous extract of Houttuynia cordata $T_{HUNB}$ on NIH 373 fibroblasts was evaluated by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetra-zoliumbromide) and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treated cells. These results were obtained as follows; The concentration of 10$^{-2}$ mg/ml of Houttuynia cordata $T_{HUNB}$ extract was shown significant antitoxic activity The number of NIH 373 fibroblasts were increased and tend to regenerate. These results suggest that Houttuynia cordata THUNB extract retains a potential antitoxic activity.oxic activity.

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Hepatoprotective Effect of Solvent Fractions from Raphiolepis indica against Oxidative Stress

  • Kim, Sun Young;Kim, Hye-Ran;Chang, Kyung-Soo
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.4
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    • pp.1269-1275
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    • 2018
  • Raphiolepis indica (R. indica) is one of evergreen shrubs belonging to the Rosaceae and is grown wildly in Jeju. This study was performed to evaluate the hepatoprotective effect of different fractions (n-hexane, dichloromethane, ethyl acetate, butanol, water) from R. indica. Anti-oxidative effects were determined using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity and total phenol contents. Hepatoprotective effect was identified by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay in Huh7 cells. Among various fractions, ethyl acetate and butanol fractions showed the lowest DPPH remained rate reaching approximately 78.7 and 65.5% at $400{\mu}g/mL$. Ethyl acetate and butanol fractions showed the total phenolic content at 164.5 and 137.3 mg GAE/g extract. The ethyl acetate and butanol fractions were resistant against oxidative stress in MTT assay and showed higher hepatoprotective effect than other fractions. Therefore, these results suggest that the ethyl acetate and butanol fractions of R. indica might have therapeutic value in liver damage.

Ex-situ conservation and cytotoxic activity assessment of native medicinal orchid: Coelogyne stricta

  • Thapa, Bir Bahadur;Thakuri, Laxmi Sen;Joshi, Pusp Raj;Chand, Krishna;Rajbahak, Sabari;Sah, Anil Kumar;Shrestha, Resha;Paudel, Mukti Ram;Park, So Young;Pant, Bijaya
    • Journal of Plant Biotechnology
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    • v.47 no.4
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    • pp.330-336
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    • 2020
  • Ex-situ conservation of the ornamental and medicinal orchid, Coelogyne stricta, was performed by mass propagation using seed culture. Propagation stages were optimized using full- and half-strength solidified MS medium with different phytohormones. Maximum seed germination (88 ± 0.5% over 6 weeks of culture) was achieved on half-strength MS medium supplemented with 15% coconut water. Maximum shoot numbers were found on full-strength MS medium supplemented with 1 mg/L BAP, 2 mg/L Kinetin, and 10% coconut water, while the longest root was developed on full-strength MS medium with 1.5 mg/L IBA. A 2:1:1 combination of coco-peat, pine bark, and sphagnum moss was found to be a suitable potting mixture resulting in 80% seedling survivability. The cytotoxic activity of extracts of both wild plants and in vitro-developed protocorms was determined using an MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay on a cervical cancer cell line. The wild plant extract inhibited the growth of 41.99% of cells, showing that this extract has moderate cytotoxic activity toward cervical cancer cells.

NF-${\kappa}B$ Activation and cIAP Expression in Radiation-induced Cell Death of A549 Lung Cancer Cells (A549 폐암세포주의 방사선-유도성 세포사에서 NF-${\kappa}B$ 활성화 및 cIAP 발현)

  • Lee, Kye Young;Kwak, Shang-June
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.5
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    • pp.488-498
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    • 2003
  • Background : Activation of the transcription factor NF-${\kappa}B$ has been shown to protect cells from tumor necrosis factor-alpha, chemotherapy, and radiation-induced apoptosis. NF-${\kappa}B$-dependent cIAP expression is a major antiapoptotic mechanism for that. NF-${\kappa}B$ activation and cIAP expression in A549 lung cancer cells which is relatively resistant to radiation-induced cell death were investigated for the mechanism of radioresistance. Materials and methods : We used A549 lung cancer cells and Clinac 1800C linear accelerator for radiation. Cell viability test was done by MTT assay. NF-${\kappa}B$ activation was tested by luciferase reporter gene assay, Western blot for $I{\kappa}B{\alpha}$ degradation, and electromobility shift assay. For blocking ${\kappa}B$, MG132 and transfection of $I{\kappa}B{\alpha}$-superrepressor plasmid construct were used. cIAP expression was analyzed by RT-PCR and cIAP2 promoter activity was performed using luciferase assay system. Results : MTT assay showed that cytotoxicity even 48 hr after radiation in A549 cells were less than 20%. Luciferas assay demonstrated weak NF-${\kappa}B$ activation of $1.6{\pm}0.2$ fold compared to PMA-induced $3.4{\pm}0.9$ fold. Radiation-induced $I{\kappa}B{\alpha}$ degradation was observed in Western blot and NF-${\kappa}B$ DNA binding was confirmed by EMSA. However, blocking NF-${\kappa}B$ using MG132 and $I{\kappa}B{\alpha}$-superrepressor transfection did not show any sensitizing effect for radiation-induced cell death. The result of RT-PCR for cIAP1 & 2 expression was negative induction while TNF-${\alpha}$ showed strong expression for cIAP1 & 2. The cIAP2 promoter activity also did not show any change compared to positive control with TNF-${\alpha}$. Conclusion : We conclude that activation of NF-${\kappa}B$ does not determine the intrinsic radiosensitivity of cancer cells, at least for the cell lines tested in this study.

Antineoplastic Effect of Several Herbal Medicine Mixtures on SNU-80 Anaplastic Thyroid Carcinoma Cell Line (수종 한약 복합물의 역형성갑상선암세포 SNU-80에 대한 항암효과)

  • Yeo, Hyun-Soo;Lee, Min-Hye;Choi, You-Kyung;Jun, Chan-Young;Park, Jong-Hyeong
    • The Journal of Internal Korean Medicine
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    • v.35 no.4
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    • pp.416-427
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    • 2014
  • Objectives: The purpose of this study was to investigate the antineoplastic effect of several herbal medicine mixtures (compositions of Astragalus membranaceu, Angelica gigas, Trichosanthes kirilowii, Panax ginseng, Rhus verniciflua Stokes) on the SNU-80 anaplastic thyroid carcinoma cell line. Methods: MTT assay was used to examine whether our herbal medicine mixtures decreased cell growth rate of SNU-80. Wound healing assay and Transwell invasion assay was performed to investigate whether our herbal medicine mixtures affect the migration and invasion of anaplastic cancer cells, SNU-80. ELISA assay was performed to know if our herbal medicine mixtures suppressed the expression of pro-invasive molecules, such as vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) secreted from SNU-80. Results: MTT assay demonstrated that A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas :T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 strongly suppressed the growth of SNU-80. Wound healing assay demonstrated that A. membranaceus:A. gigas=3:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the migration of SNU-80. Transwell invasion assay demonstrated that A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii =1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng :R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the invasion of SNU-80. ELISA assay demonstrated that A. membranaceus :A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 suppressed the expression of VEGF. Also, A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus :A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes =1:1:1:1:1 or 3:1:1:1:1 suppressed the expression of MMP-2. Conclusions: The results obtained in this study suggest that several herbal medicine mixtures suppresse the growth and inhibit the migration and invasion of SNU-80, which is anaplastic thyroid cancer cells. Especially, A. membranaceus:A. gigas: T. kirilowii=1:1:1 mixture had a stronger anti-cancer effect.

Protective Effect of Prunella spica Extracts against H2O2-Induced Cytotoxicity in PC12 Cells (Hydrogen peroxide가 유도하는 세포독성으로부터 PC12 세포를 보호하는 하고초(Prunella spica) 추출물의 영향)

  • Kim, Hyun-Jung;Lee, Jeung-Min;Moon, Seong-Hee;Park, Hae-Ryong
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1121-1126
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    • 2010
  • The oxidative stress induced by reactive oxygen species (ROS) may play an important role in the pathogenesis of neurodegenerative diseases. In this study, we investigated the neuroprotective effects of methanolic extracts of Prunella Spica (PSE) against $H_2O_2$-induced oxidative stress in PC12 cells. The cells exposed to $H_2O_2$-induced oxidative stress were treated with various concentrations of PSE; this treatment resulted in the induction of a dose-dependent protective effect, which was evidenced by the results of MTT reduction assay, lactate dehydrogenase (LDH) release assay, morphological assay, and colony-formation assay. Interestingly, we also observed reduction of apoptotic bodies in the Hoechst staining and flow cytometric analysis. These data show that apoptosis was significantly suppressed in the PC12 cells that were exposed to $H_2O_2$-induced oxidative stress and treated with PSE. These results suggest that Prunella Spica could be a new potential protective agent against $H_2O_2$-induced oxidative stress.

Antineoplastic Effect of Several Herbal Medicines on SNU-80 Anaplastic Thyroid Carcinoma Cell Line (황기, 당귀, 칠피, 천화분의 역형성갑상선암세포 SNU-80에 대한 항암효과)

  • Yeo, Hyun Soo;Lee, Min Hye;Ko, Seong Gyu;Choi, You Kyung;Jun, Chan Young;Park, Jong Hyeong
    • Journal of Society of Preventive Korean Medicine
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    • v.18 no.1
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    • pp.83-92
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    • 2014
  • Objective : This study was performed to investigate the antineoplastic effect of Astragalus membranaceus, Angelica gigas, Rhus verniciflua Stokes and Trichosanthes kirilowii on SNU-80 anaplastic thyroid carcinoma cell line. Method : We examined whether our herbal medicines decreases cell growth rate of SNU-80 using MTT assay. We performed western blot analysis to verify that our herbal medicines induces apoptosis via caspase-dependent mechanism. We also performed wound healing assay and transwell invasion assay to investigate whether our herbal medicines affects the migration and invasion of anaplastic cancer cells, SNU-80. We also carried out ELISA assay to know our herbal medicines suppresses the expression of proinvasive molecules, such as VEGF and MMP-2 secreted from SNU-80. Results : MTT assay demonstrates that Angelica gigas, Rhus verniciflua Stokes, and Trichosanthes kirilowii suppressed strongly the growth of SNU-80. Western blot analysis demonstrates that Trichosanthes kirilowii induces apoptosis activating the cleavages of caspases (caspase-8, caspase-3) and PARP. Wound healing assay demonstrates that Rhus verniciflua Stokes, and Trichosanthes kirilowii inhibited the migration of SNU-80. Transwell invasion assay demonstrates that Rhus verniciflua Stokes, and Trichosanthes kirilowii inhibited the invasion of SNU-80. Elisa assay demonstrates that Astragalus membranaceus, Angelica gigas, Rhus verniciflua Stokes, and Trichosanthes kirilowii suppressed the expression of VEGF and MMP-2. Conclusion : We could conclude that several herbal medicines suppresses the growth and inhibits the migration and invasion of SNU-80 which is anaplastic thyroid cancer cells. Especially, Rhus verniciflua Stokes, Trichosanthes kirilowii had stronger anti-cancer effect suggesting that we can apply them to treat anaplastic thyroid cancer.

Cytotoxic Evaluation of the Ethyl Acetate Soluble Fraction of Perilla frutescens (소엽 Ethyl acetate 분획의 세포독성 평가)

  • Kim Sung-Eun;Chun Hyun-Ja;Kim Il-Kwang;Han Du-Seok;Lee Hyun-Ok;Ahn Jong-Woong;Lee Mee-Hee;Baek Seung-Hwa
    • Journal of Society of Preventive Korean Medicine
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    • v.5 no.2
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    • pp.106-113
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    • 2001
  • The Cytotoxic activity of the ethyl acetate soluble fraction of Perilla frutescens on human oral epithelioid carcinoma cell lines was evaluated by 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl-2H-tetrazalium bromide (MTT) colorimetric method. The light microscopic study was carried out to observe the morphological changes of cultured human oral epitheloid carcinoma cell lines (KB) These results were obtained as follows; 1. subfraction 1 of the ethyl acetate soluble fraction of perilla frutescens was shown significant cytotoxic activity (p<0.001) at 10-2 mg/ml concentration, this concentration was decreased the cytotoxic activity of 70.3% . 2. The comparison of IC50 values of various subfractions in human oral epithelioid carcinoma cell lines was exhibited that their susceptibility to these subfractions decreased in the following order ; subfraction 5 > subfraction 4 > subfraction 3 > subfraction 2 > subfraction 1 by MTT assay. In light microscopy, the subfraction 1 of the ethyl acetate soluble fraction of Perilla frutescens showed the highest cytotoxic activity. These findings suggest that subfraction 1 possessed the most cytotoxic constituents.

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