• Title/Summary/Keyword: MTT reduction assay

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In vitro Cytotoxic Effect of Extracts from Styela plicata (오만둥이(Styela plicata) 추출물의 in vitro 세포독성 효과)

  • Lee, Bo-Bae;Cha, Mi-Ran;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.9
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    • pp.1099-1105
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    • 2007
  • The present study describes the preliminary evaluation of the anticancer activity of Styela plicata. Freeze-dried S. plicata was extracted with methanol, ethanol, acetone, and water, and then anticancer effect of the extracts was measured by the MTT reduction assay and phase-contrast microscopy on the HT-29 human colon carcinoma cells. Among the extracts, acetone extract showed the highest anticancer activity. The cell proliferation rates markedly decreased by 94.0% at the concentration of 500 ${\mu}g/mL$ of acetone extract compared with control cells. The acetone extract was further fractionated with hexane, diethyl ether, ethyl acetate, and water layer according to the degree of polarity. The HT-29 cells with hexane layer extract (250 ${\mu}g/mL$) decreased the cell viability to 5.1% of untreated control. The growth of SW620, HeLa, and MCF-7 cells was decreased to about 10%, by the treatment of hexane layer extract 250 ${\mu}g/mL$. Theses results suggest extracts from S. plicata as possible natural cancer therapeutic material.

Neuroprotective Effect of Extracts from Root Bark of Morus alba on Glutamate-induced Cytotoxicity in Neuronal Cells. (Glutamate가 유도하는 세포독성으로부터 신경세포를 보호하는 상백피 추출물의 효과)

  • Kim, Hyun-Jung;Kim, Ji-Hyun;Son, Eun-Soon;Lee, Jeung-Min;Park, Hae-Ryong
    • Journal of Life Science
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    • v.19 no.7
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    • pp.963-967
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    • 2009
  • This study evaluated the neuroprotective effect of extracts from the root bark of Morus alba (MA) against glutamate-induced cytotoxicity in neuronal cells. Glutamate-induced cytotoxicity was shown by MTT reduction assay. The neuroprotective effects of methanol, ethanol, and acetone extracts from MA against glutamate-induced cytotoxicity were measured. Among the three extracts, the methanolic extracts showed the highest protective effect, as determined by the results of an morphological assay, a lactate dehydrogenase release assay. Furthermore, the methanol extracts were fractionated sequentially with hexane, diethyl ether, ethyl acetate, and water layer according to degree of polarity. The hexane fractions exhibited a neuroprotective effect against glutamate-stressed N18-RE-105 cells. Therefore, these results suggest that extracts of MA could be a new potential candidate as a protective substance against glutamate-induced cytotoxicity.

Protective Effect of Methanolic Extracts from Dendrobium nobile Lindl. on $H_{2}O_{2}$-induced Neurotoxicity in PC12 cells (석곡 MeOH 추출물이 $H_{2}O_{2}$에 의한 신경세포 보호효과에 미치는 영향)

  • Yoon, Mi-Young;Kim, Ju-Young;Hwang, Ji-Hwan;Cha, Mi-Ran;Lee, Mi-Ra;Jo, Kyung-Jin;Park, Hae-Ryong
    • Applied Biological Chemistry
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    • v.50 no.1
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    • pp.63-67
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    • 2007
  • The neuroprotective effect of methanolic extracts from Dendrobium nobile Lindl. (DME) against $H_{2}O_{2}$-induced neurotoxicity in PC12 cells was investigated. The treatment of PC12 cells with various DME concentrations under $H_{2}O_{2}$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay, an LDH release assays, and a morphological assay. Interestingly, we also detected reduction of apoptotic bodies and inhibition of caspase-3 activity by DME in $H_{2}O_{2}$-indeced PC12 cells. These data show that the neuroprotective effect of DME against PC12 cells might be related to the suppression of caspase-3 activation. Therefore, these results suggest that DME could be a new potential candidate as chemotherapeutic agents against neuronal diseases.

Evaluation of Whitening Activity and Wrinkle Inhibitory Effect of Ethanol Extracts of Nelumbinis Rhizomatis Nodus (우절 에탄올추출물의 미백활성능과 주름저해 효능평가)

  • Jang, Young-Ah;Yeom, Bo-Seul;Kim, Se-Gie;Lee, Jin-Tae
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1192-1199
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    • 2019
  • The intention of this study was to confirm the possible use of an ethanol extracts of Nelumbinis Rhizomatis Nodus (NRN) as a cosmetic material. To this end, we extracted NRN with 70% ethanol and performed biological activity evaluation of whitening efficacy and wrinkle reduction. We performed cellular tyrosinase inhibition and melanin contents assay to check the whitening activity of NRN and carried out a toxicity evaluation of NRN via an MTT assay and the amounts of associated proteins that affect melanin production in a melanoma cell line (B16F10). And collagenase inhibitory assay was performed for the evaluation of anti-wrinkle of samples. In addition, a toxicity evaluation using an MTT assay and matrix metalloprotease (MMP-1) and procollagen synthesis inhibition by NRN were evaluated in a fibroblast cell line (CCD-986sk). Western blot results for the whitening activity evaluation revealed that the levels of two proteins related to melanin production, tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2), were decreased in a dose-dependent manner. Moreover, collagenase inhibition activity at a concentration of $500{\mu}g/ml$ NRN by measuring epigallocatechin-3-gallate (EGCG) was increased by more than 80% compared to the control group. Meanwhile, procollagen synthesis was reduced by 68.8% in the UVB-induced CCD- 986sk cells group whereas collagen synthesis recovered by 80.2% with $25{\mu}g/ml$ NRN. The MMP-1 expression rate showed 20.2% reduction at $25{\mu}g/ml$. The results of the experiments verified the whitening and wrinkle suppression effects of NRN and confirmed that it could be used as a safe natural cosmetic material in the future.

The Flavin-Containing Reductase Domain of Cytochrome P450 BM3 Acts as a Surrogate for Mammalian NADPH-P450 Reductase

  • Park, Seon-Ha;Kang, Ji-Yeon;Kim, Dong-Hyun;Ahn, Taeho;Yun, Chul-Ho
    • Biomolecules & Therapeutics
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    • v.20 no.6
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    • pp.562-568
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    • 2012
  • Cytochrome P450 BM3 (CYP102A1) from Bacillus megaterium is a self-sufficient monooxygenase that consists of a heme domain and FAD/FMN-containing reductase domain (BMR). In this report, the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) by BMR was evaluated as a method for monitoring BMR activity. The electron transfer proceeds from NADPH to BMR and then to BMR substrates, MTT and CTC. MTT and CTC are monotetrazolium salts that form formazans upon reduction. The reduction of MTT and CTC followed classical Michaelis-Menten kinetics ($k_{cat}=4120\;min^{-1}$, $K_m=77{\mu}M$ for MTT and $k_{cat}=6580\;min^{-1}$, $K_m=51{\mu}M$ for CTC). Our continuous assay using MTT and CTC allows the simple, rapid measurement of BMR activity. The BMR was able to metabolize mitomycin C and doxorubicin, which are anticancer drug substrates for CPR, producing the same metabolites as those produced by CPR. Moreover, the BMR was able to interact with CYP1A2 and transfer electrons to promote the oxidation reactions of substrates by CYP1A2 and CYP2E1 in humans. The results of this study suggest the possibility of the utilization of BMR as a surrogate for mammalian CPR.

Antioxidant Activity and Neuroprotective Effect of Psoralea corylifolia Linne Extracts (파고지 (Psoralea corylifolia Linne) 추출물의 항산화 작용에 의한 신경세포 보호효과)

  • Yoon, Mi-Young;Lee, Bo-Bae;Kim, Ju-Young;Kim, Yong-Seong;Park, Eun-Ju;Lee, Seung-Cheol;Park, Hae-Ryong
    • Korean Journal of Pharmacognosy
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    • v.38 no.1
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    • pp.84-89
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    • 2007
  • The present study describes the preliminary evaluation of the antioxidant activities and the neuroprotective effect of methanolic extracts from Psoralea corylifolia Linne (PCE). The antioxidant activities and neuroprotective effect of the PCE were evaluated by total phenolic contents (TPC), DPPH radical scavenging activity (RSA), reducing power (RP), MTT reduction assay, and LDH release assay. TPC, DPPH RSA, and RP of the extract at concentration of 100 ${\mu}g$ was 125.93 ${\mu}g$, 63.81%, 0.138, respectively, and those were concentration dependent. The treatment of PC12 and N18-RE-105 cells with various PCE concentrations under $H_2O_2$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay and LDH release assay. Therefore, these results suggest that PCE could be a new potential candidate as an antioxidant against neuronal diseases.

Neuroprotective Effects of Extracts from Diospyros kaki L. Peel (단감(Diospyros Kaki L.)껍질 추출물의 신경세포 보호 효과)

  • Lee, Mi-Ra;Moon, Seong-Hee;Choi, Ae-Ran;Lee, Seung-Cheol;Ahn, Kwnag-Hwan;Park, Hae-Ryong
    • Korean journal of food and cookery science
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    • v.27 no.4
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    • pp.67-73
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    • 2011
  • This study was performed to assess the neuroprotective effects of methanolic extracts from sweet persimmon peel (PPE) against glutamate-induced cytotoxicity in hybridoma N18-RE-105 cells. The neuroprotective effects of PPE in N18-RE-105 cells were measured using the MTT reduction assay, LDH release assay, and phase-contrast microscopy. The results of the MTT reduction assay showed that treating cells with 500 ${\mu}g/ml$ PPE resulted in cell viability of 66.9%. Additionally, the morphological changes and the results of the LDH release assay showed that glutamate-induced damage to nerve cells was strongly inhibited by PPE. GSH content of N18-RE-105 cells was 3.5 ${\mu}M$ compared to that of the control, whereas pretreatment with 500 ${\mu}g/ml$ PPE increased GSH content by 4.7 ${\mu}M$. PPE was fractionated with hexane, and that layer had the highest neuroprotective effects in glutamate-stressed N18-RE-105 cells. In conclusion, our data showed that glutamate potentiated the effects of N18-RE-105 cell death by a mechanism involving oxidative stress. Therefore, PPE may be a potential candidate for prevention and therapy of neurodegenerative diseases.

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.

Antioxidant Effects of Psoraleae Fructus in GC-1 Cells (보골지(補骨脂)가 남성 생식세포 GC-1의 항산화에 미치는 영향)

  • Oh, Myung-Sook;Kim, Do-Rim;Kim, So-Yeon;Chang, Mun-Seog;Park, Seong-Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.1
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    • pp.81-86
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    • 2005
  • The purpose of this study is to examine the antioxidant activity in the germ cells of the extract of Psoraleae fructus. The extract was studied for dipheny-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay and lipid perixidation by malondialdehyde (MDA) formation, respectively. The results showed that the extract scavenged DPPH radical with the IC50 being 0.427 mg/mL. The extract was dose-dependent in growth of GC-1 cell. $H_2O_2$-induced cytotoxicity (67.7 %) was blocked by the extract concentration- dependently. Furthermore, the extract also displayed a dose-dependent reduction of MDA formation on $H_2O_2$-induced lipid peroxidation. In conclusion, the extract of Psoraleae fructus has potent antioxidant activity.

Antioxidant Effects of Corni Fructus in GC-1 Cells (산수유(山茱萸)가 남성 생식세포 GC-1의 항산화에 미치는 영향)

  • Oh Myung-Sook;Kim Do-Rim;Sung Eun-Jin;Chang Mun-Seog;Park Seong-Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.6
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    • pp.1541-1545
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    • 2005
  • The purpose of this study is to examine the antioxidant activity in the germ cells of the extract of Corm fructus. The extract was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay and lipid perixidation by malondialdehyde (MDA) formation, respectively. The results showed that the extract scavenged DPPH radical with the IC50 being $200{\mu}g/mL$. The extract at concentrations of $10-500{\mu}g/ml$ showed dose-dependent in growth of GC-1 cell. $H_2O_2$-induced cytotoxicity (63.0%) was blocked by the extract (10, 50, 100, 250 and $500{\mu}g/ml$) concentration-dependently. Furthermore, the extract (50, 100 and $250{\mu}g/ml$) also displayed a dose-dependent reduction of MDA formation on $H_2O_2$-induced lipid peroxidation. In conclusion, the extract of Corm fructus has potent antioxidant activity.