• Title/Summary/Keyword: MTT assay

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생체외 모델에서 아우레오바지디움 유래 베타 글루칸이 피부창상 치유에 미치는 영향 (Effect of ${\beta}$-glucan Originated from Aureobasidium on the Dermal Wound Repair in Vitro Model)

  • 이정우;권영삼;장광호
    • 한국임상수의학회지
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    • 제30권6호
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    • pp.403-408
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    • 2013
  • 본 연구의 목적은 in vitro 실험에서 human dermal fibroblast cell을 이용한 섬유아세포 증식과 콜라겐 생성을 통한 전층피부창상치유에 대한 효과를 알아보는 것이다. In vitro 실험에서, 베타글루칸 및 양성대조물질 oncostatin M을 사용하여 처치 후 24 또는 48시간 이후 MTT assay와 세포수 관찰을 통해 섬유모세포 증식효과에 대해 평가하였다. Procollagen 생산에 대한 효과를 평가하기 위해 베타글루칸 및 양성대조물질 oncostatin M 처치 48시간 후 hydroxyproline을 HPLC를 이용하여 정량하여 평가하였다. 창상재구축에 대한 효과를 평가하기 위해 베타글루칸 및 양성대조물질 fucoidan fraction 7을 FPCL 모델을 이용하여 평가하였다. 베타글루칸 처치군은 음성대조군 대비 0.1 mg/ml 이상의 농도에서 용량의존적이고 유의적인 OD 또는 섬유아세포수의 증가가 관찰되었다. 게다가 베타글루칸 처치군은 창상면으로 유주된 세포수가 음성대조군 대비 1 mg/ml 이상의 농도에서 용량의존적이고 유의적인 증가가 관찰되었다. 그러나 베타글루칸 처치군은 음성대조군 대비 모든 용량에서 procollagen 생산에 대한 유의적인 변화가 관찰되지 않았다.

Purification of Cucurbitacins D, E, and I from Ecballium Elaterium (L.) A. Rich Fruits and Study of Their Cytotoxic Effects on the AGS Cell Line

  • Jafargholizadeh, Naser;Zargar, Seyed Jalal;Yassa, Narguess;Tavakoli, Saeed
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권10호
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    • pp.4631-4635
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    • 2016
  • Background: The plant Ecballium elaterium (L.) A. Rich, belongs to the Cucurbitaceae family which occupies an important position in traditional medicine prescriptions. It has been reported that a freeze-dried aqueous extract of E. elaterium fruits has cytotoxic effects on the AGS human stomach adenocarcinoma cell line. We here focused on anticancer effects of the main chemicals purified from E. elaterium fruits. Materials and Methods: We isolated cucurbitacins D, E, and I from chloroform, and ethyl acetate fractions of a methanolic extract of E. elaterium fruits and assessed their cytotoxic effects on the AGS cell line by MTT assay. The methanolic extract was fractionated to petroleum ether, chloroform, and ethyl acetate fractions. The compounds isolated by column chromatography were identified by NMR spectroscopy. Results: After 24 h of incubation with AGS cells, the IC50 values were 0.3, 0.1, and $0.5{\mu}g/ml$ for cucurbitacins D, E, and I respectively. Conclusions: This finding suggests that because of its cucurbitacins, E. elaterium fruit may have some cytotoxic effects on gastric cancer cells. Also, compared with D and I, cucurbitacin E showed greater potency in this regard.

HepG2 세포에서 tert-butyl hydroperoxide로 유도된 간독성에 대한 kurarinone의 세포 보호 효과 (Cytoprotective effects of kurarinone against tert-butyl hydroperoxide-induced hepatotoxicity in HepG2 Cells)

  • 김상찬;이종록;박숙자
    • 대한한의학방제학회지
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    • 제26권3호
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    • pp.251-259
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    • 2018
  • Objective : Kurarinone is one of the flavonoids isolated from Sophorae Radix with various biological activities including anti-microbial effect. In this study, we investigated the effects of Kurarinone on tert-butyl hydroperoxide (tBHP)-induced oxidative stress finally leading to apoptosis in human hepatoma cell line HepG2. Methods : To determine the effects on cell viability, the cells were exposed to tBHP ($100{\mu}mol/l$) after pretreatment with kurarinone (0.5 and $1{\mu}g/ml$). Cell viability was measured by MTT assay. To reveal the possible mechanism of cytoprotectivity of kurarinone, levels of reactive oxygen species, intracellular glutathione, mitochondrial membrane potential, and expression of caspase were examined. Results : tBHP-induced cell death was due to oxidative stress and the resulting apoptosis. Kurarinone dose-dependently protected cells from apoptosis when determined by MTT and TUNEL assay. Consistent with this observation, decreased expression of pro-caspase 3/9 protein by tBHP was restored by kurarinone. Kurarinone also showed anti-oxidative effects by inhibiting generation of ROS and depletion of GSH in tBHP-stimulated HepG2 cells. In addition, kurarinone significantly recovered disruption of mitochondrial membrane potential (MMP) as a start sign of hepatic apoptosis induced by oxidative stress. Conclusion : From these results, it was concluded that kurarinone protected tBHP-induced hepatotoxicity with anti-oxidative and anti-apoptotic activities. Our results suggest that kurarinone might be beneficial to hepatic disorders caused by oxidative stress.

동물 세포 증식을 위한 저출력 광 소스의 특성 (Characteristics of Low-level Light Source for Animal Cell Proliferation)

  • 천민우;김성환;송창훈;문성표;김태곤;박용필;김대영;김영수
    • 한국전기전자재료학회논문지
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    • 제20권1호
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    • pp.92-97
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    • 2007
  • This paper performed the basic study for developing the Photodynamic Therapy Equipment for medical treatment. We developed the equipment palpating cell proliferation using a high brightness LED. This equipment was fabricated using a micro-controller and a high brightness LED, and designed to enable us to control light irradiation time, intensity, frequency and so on. Especially, to control the light irradiation frequency, FPGA was used, and to control the change of output value, TLC5941 was used. Control stage is divided into 30 levels by program. Consequently, the current value could be controlled by the change of level in Continue Wave(CW) and Pulse Width Modulation(PWM), and the output of a high brightness LED could be controlled stage by stage. And then, each experiment was performed to irradiation group and non-irradiation group for both Rat bone marrow and Rat tissue cells. MTT assay method was chosen to verify the cell increase of two groups and the effect of irradiation on cell proliferation was examined by measuring 590 nm transmittance of ELISA reader. As a result, the cell increase of Rat bone marrow and tissue cells was verified in irradiation group as compared to non-irradiation group. The fact that specific wavelength irradiation has an effect on cell vitality and proliferation is known through this study.

쑥부쟁이 분획물의 in vitro 암세포증식 억제 및 QR 유도효과 (Inhibitory Effects on Cell Survival and Quinone Reductase Induced Activity of Aster yomena Fractions on Human Cancer Cells)

  • 정복미;임상선;박윤자;배송자
    • 한국식품영양과학회지
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    • 제34권1호
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    • pp.8-12
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    • 2005
  • 본 연구는 약용 및 식용 식물로 사용되고 있는 쑥부쟁이 추출물과 분획물의 암세포 증식 억제 효과와 QR유도 효과를 실험하였다. 쑥부쟁이의 metanol 추출 및 용매 분획물의 암세포 증식억제 효과를 MTT assay로 실험한 결과, 3종의 인체 암세포주, 간암세포주인 HepG2, 자궁경부암 세포주인 HeLa 및 유방암 세포주인 MCF-7에서 모두 쑥부쟁이의 ethyl ether 분획층인 AYMEE층과 ethyl acetate 분획층인 AVMEA층에서 암세포 성장 저지 효과가 제일 컸다. 특히 HepG2 세포주에서 는 AYMEE를 200 $\mu$g/mL 첨가시켰을 때 94.5%의 암세포 증식 억제 효과가 나타났으며, 농도를 증가시킬수록 그 효과가 커져 500 $\mu$g/mL에서는 97.7%의 저지 효과를 보였다. MCF-7 세포주와 HeLa 세포주에서도 이와 비슷한 경향이었다. Butanol층인 AYMB와 메탄올층인 AYM에서는 대체적으로 낮은 농도에서는 효과가 약하였으나 300 $\mu$g/mL 이상에서는 높은 암세포 성장저지 효과를 보였다. HepG2 세포를 이용하여 암예방 지수인 QR 유도 활성을 측정한 결과, 다른 분획층에 비해 비극성 용매층인 hexane 분획층, AYMH에서 QR 유도 활성을 증가시키는 경향이었다.

봉독이 위암 세포주에 미치는 효과 (Effects of the Bee Venom on Human Gastric Adenocarcinoma Cell Lines)

  • 허경;김명호;임성우
    • 동의생리병리학회지
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    • 제27권1호
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    • pp.92-98
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    • 2013
  • Bee Venom(below BV) has been used in alternative medicine to treat the diseases, such as pain diseases. BV contains a variety of peptides, including melittin, apamin, adolapin, MCD peptide, enzymes(i.e. PLA2), amines(i.e. histamine and epinephrine), and nonpeptide components. The two main components of BV are melittin and PLA2. The cell cytotoxic effects through the activation of PLA2 by melittin have been suggested to be the critical mechanism for the depress of cancer cell. Melittin and PLA2 have been reported to induce apoptosis and to possess anti-cancer effects and neurite outgrowth in PC12 cells. Analysis of proliferation was confirmed by MTT assay. BV decreased cell number through dose- and duration-dependent manner and these effects are apparent at a concentration of 3 ${\mu}g/ml$. To observe which signaling molecules will be activated by BV, phosphorylation of ERK, p38 MAPK, JNK and ERM were examined by Western blot analysis. To study the long term effect of BV in human gastric adenocarcinoma cell lines, the image of cells treated with BV for 4 days were obtained. BV was shown to exhibit anti-cancer activity in human gastric adenocarcinoma cell lines at a broad range of concentrations of 3 ${\mu}g/ml$. ERK, p38 MAPK and JNK were found to increase in BV treated cells. However, ERM which known to be involved in the cell death, was gradually decreased to 30minutes after addition 3 ${\mu}g/ml$ of BV. These results provide a possible BV-induced inhibitory signal for cancer proliferation that is initiated by the decrease in ERM activity. Moreover, it is likely that the activation of ERK, p38 MAPK and JNK are required for the BV-induced inhibition of cancer proliferation.

Low LET X-ray가 췌장 ${\beta}$ 세포와 신경세포에 미치는 효과 (Effect on Pancreatic Beta Cells and Nerve Cells by Low LET X-ray)

  • 박광훈;김구환
    • 대한방사선기술학회지:방사선기술과학
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    • 제37권1호
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    • pp.21-28
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    • 2014
  • 배양된 췌장베타세포와 신경세포를 10% FBS (fetal bovine serum), 11.1 mM glucose의 normal 조건과 1% FBS, 30 mM glucose의 hyperglycemia 조건으로 배양하고, 저 LET X-선을 0.5 Gy/hr의 선량률로 총 선량이 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5 Gy가 되도록 저 LET 방사선을 조사한 후 MTT assay로 생존율을 측정하고 비교하였다. 분화된 췌장베타세포에 방사선을 조사하지 않았을 때는 normal 조건에 비해 hyperglycemia 조건의 생존율이 경미한 감소를 보였다. 그러나 X선의 총 선량이 점차 누적됨에 따라 normal 조건에서 생존율이 조금씩 감소하는 것에 비해 hyperglycemia 조건에서는 급격히 감소하여 시너지 효과를 나타냈다. 미분화된 신경세포에서는 방사선을 조사하지 않은 때도 normal 조건에 비해 hyperglycemia 조건의 생존율이 뚜렷하게 감소하였다. X선의 총 선량이 점차 누적됨에 따라 normal 과 hyperglycemia 조건 모두에서 비교적 급격한 세포사멸이 나타났지만 그 생존율의 감소비율이 거의 평행하게 진행되어 시너지 효과를 보이지 않았다.

여정실 물 분획물이 인체 진피 섬유아세포의 증식에 미치는 영향 (Effect of Fructus Ligustri Lucidi $H_2O$ Extract on Cell Proliferation in Hman Dermal Fibroblast)

  • 임난영;김대성;고경숙;문연자;우원홍
    • Korean Journal of Acupuncture
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    • 제28권3호
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    • pp.43-51
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    • 2011
  • Objectives : In this study, we investigated the effect of Fructus Ligustri Lucidi $H_2O$ fraction (FLLW) on cell proliferation, and the phosphorylation of ERKs and Akt in human dermal fibroblast neonatal (HDFn). Methods : After treatment of HDFn with FLLW, MTT assay was performed to quantitatively determine cellular viability. The ERK and Akt pathways were analyzed in vitro by Western blot in a HDFn. HDFn proliferation after FLLW and minoxidil treatment in the absence or presence of PD98059, a MEK inhibitor, LY294002, and a PI3K inhibitor, was examined by Western blot or MTT assay. Results : FLLW increased cell proliferation in a dose-dependent manner and minoxidil used as positive control also induced cell proliferation in HDFn. FLLW increased the phosphorylation of ERK and Akt. In addition, minoxidil, too, induced the phosphorylation of ERK and Akt in HDFn. PD98059 and LY294002 significantly attenuated FLLW-inducible p-ERK and p-Akt expression and proliferation in cultured HDFn. Conclusions : Our results suggest that FLLW stimulates the growth of fibroblast cells through ERK and Akt pathways. Therefore, FLLW is a potential agent for the inducer of fibroblast growth.

L1210 및 $P388D_1$에 대한 고삼 추출물의 세포독성에 관한 연구(II) (Studies on the Cytotoxicity of Sophora flavescens Ait. Extract Against L1210 and $P388D_1$ Cells (II))

  • 조훈;양은영;김종수;유일수;류도곤;강길웅;이정호;백승화
    • 생약학회지
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    • 제30권4호
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    • pp.351-354
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    • 1999
  • This study was carried out to evaluate cytotoxicity of the extracts from Sophora flavescens Ait. against L1210 (lymphocytic leukemia) and $P388D_1$ (lymphoid neoplasms) cells in vitro. We have determined cytotoxicity by MTT {3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazo-liumbromide} assay. The order of cytotoxicity of Sophora flavescens Ait. extracts against L1210 and $P388D_1$ cells in vitro is as follows: AM> EASF > CFSF > MTSF > WSF > HXSF and AM> EASF> CFSF> MTSF> HXSF> WSF. These results suggest that the ethyl acetate soluble extract of Sophora flavescens Ait may be a valuable choice for the development of antitumor agents.

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천화분의 카드뮴에 대한 독성억제효과(III) (The Inhibitory Effects of Trichosanthes kirilowii Root against Cadmium Induced Cytotoxicity (III))

  • 이정호;유일수;김신기;이기남;정우영;한두석;백승화
    • 생약학회지
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    • 제32권1호통권124호
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    • pp.15-21
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    • 2001
  • This study was conducted to investigate the antitoxic components in the water extract of the roots of Trichosanthes kirilowii (Cucurbitaceae). The results were as follows: Generally, detoxication effects by the water extract of T. kirilowii increased in proportion to the concentrations. Experimental animals were treated with cadmium and T. kirilowii water extract by oral administration. When 40 mg/kg dosage of T. kirilowii extract was administrated it showed the highest antitoxic effects in metallothionein induction. After the water extract treatment, body weights did not increase in proportion to the extract concentrations. These results suggest that T. kirilowii extract increased metallothionein concentration and decreased the toxicity of cadmium in rats. In vitro the antitoxic activity of water extract of T. kirilowii on NIH 3T3 fibroblasts was evaluated by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) 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 T. kirilowii extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that T. kirilowii extract retains a potential antitoxic activity.

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