• Title/Summary/Keyword: MTT assay

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

  • Lee, Jung-Woo;Kwon, Young-Sam;Jang, Kwang-Ho
    • Journal of Veterinary Clinics
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    • v.30 no.6
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    • pp.403-408
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    • 2013
  • The objective of the present study is to detect the effect of ${\beta}$-glucan originated from Aureobasidium on the proliferation and collagen production in human dermal fibroblast cells with wound repopulation in vitro. The proliferative effects were assessed using a MTT assay as well as cell counts at 24 and 48 hr after treatment. Hydroxyproline was measured as an index of procollagen production with reverse-phase high pressure liquid chromatography. Oncostatin M was used as a reference agent. In glucagon treated group, dose-dependent and significant increase of optical density or fibroblast cell numbers was demonstrated, when compared with those of control from 0.1 mg/ml concentration. In addition, the numbers of cells which had migrated into the wound defects were more significantly and dose-dependently increased than those of non-treated control. However, no meaningful effects on the procollagen production were observed.

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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    • v.17 no.10
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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.

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

  • Kim, Sang Chan;Lee, Jong Rok;Park, Sook Jahr
    • Herbal Formula Science
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    • v.26 no.3
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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.

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

  • Cheon, Min-Woo;Kim, Seong-Hwan;Song, Chang-Hun;Mun, Seong-Pyo;Kim, Tae-Gon;Park, Yong-Pil;Kim, Dae-Young;Kim, Young-Su
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.20 no.1
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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.

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

  • Jung, Bok-Mi;Lim, Sing-Sn;Park, Yun-Ja;Bae, Song-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.1
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    • pp.8-12
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    • 2005
  • This study was performed to determine the inhibitory effects on cell survival and Quinone reductase induced activity of Aster yomena (AY) on human cancer cells which, using methanol, was extracted and fractionated into five different solvent types: hexane (AYMH), ethylether (AYMEE), ethylacetate (AYMEA), butanol (AYMB) and aqueous (AYMA) partition layers. The experiment was conducted to determine cytotoxicity of various Aster yomena partition layers on HepG2, HeLa and MCF-7 cells by MTT assay. Among various partition layers of Aster yomena, A YMEE and A YMEA showed the strong cytotoxic effects on all cancer cell lines we used. The Quinone reductase (QR) induced activity on HepG2 cells, A YMH at a does of 100 $\mu$g/mL was 2.46 times more effective compared to the control value of 1.0.

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

  • Heo, Gyeong;Kim, Myung Ho;Lim, Seong Woo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.1
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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.

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

  • Park, Kwang-Hun;Kim, Kgu-Hwan
    • Journal of radiological science and technology
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    • v.37 no.1
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    • pp.21-28
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    • 2014
  • Cultured pancreatic beta cells and nerve cells, it is given normal condition of 10% FBS (fetal bovine serum), 11.1 mM glucose and hyperglycemia codition of 1% FBS, 30 mM glucose. For low LET X-ray irradiated with 0.5 Gy/hr dose-rate(total dose: 0.5 to 5 Gy). Survival rates were measured by MTT assay. When non irradiated, differentiated in the pancreatic beta cells experiment is hyperglycemia conditions survival rate compared to normal conditions survival rate seemed a small reduction. However increasing the total dose of X-ray, the survival rate of normal conditions decreased slightly compared to the survival rate of hyperglycemia conditions, the synergistic effect was drastically reduced. When non irradiated, undifferentiated in the nerve cells experiment is hyperglycemia conditions survival rate compared to normal conditions survival rate seemed a large reduction. As the cumulative dose of X-ray normal conditions and hyperglycemia were all relatively rapid cell death. But the rate of decreased survivals by almost parallel to the reduction proceed and it didn't show synergistic effect.

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

  • Lim, Nan-Young;Kim, Dae-Sung;Ko, Kyung-Sook;Mun, Yeun-Ja;Woo, Won-Hong
    • Korean Journal of Acupuncture
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    • v.28 no.3
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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.

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

  • Cho, Hoon;Yang, Eun-Yeong;Kim, Jong-Soo;Yoo, Il-Soo;Ryu, Do-Gon;Kang, Kil-Ung;Lee, Jeong-Ho;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.30 no.4
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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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The Inhibitory Effects of Trichosanthes kirilowii Root against Cadmium Induced Cytotoxicity (III) (천화분의 카드뮴에 대한 독성억제효과(III))

  • Lee, Jeong-Ho;You, Il-Soo;Kim, Shin-Kee;Lee, Ki-Nam;Chung, Woo-Young;Han, Du-Seok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.32 no.1 s.124
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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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