• Title/Summary/Keyword: MTT assay

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The Inhibitory Effects of the Methanol Extract of Houttuynia cordata $T_{HUNB}$ against Cadmium Induced Cytotoxicity (V) (어성초 메탄올 추출물의 카드뮴에 대한 독성억제효과(V))

  • Lee, Jeong-Ho;Jeong, Seung-Il;You, Il-Soo;Kim, Shin-Kee;Lee, Ki-Nam;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.61-67
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    • 2001
  • This study was conducted to investigate the antitoxic agent in methanol extract of Houttuynia cordata $T_{HUNB}$. Detoxication effects By H. cordata $T_{HUNB}$ extract increased in proportion to the extract concentrations. When 40 mg/kg dosage of H. cordata $T_{HUNB}$ extract was administered, it showed the highest antitoxic effects in metallothionein induction. After the extract treatment, body weights generally increased in proportion to the extract concentrations. From the above results, H. cordata $T_{HUNB}$ extract increased metallothionein concentrations and decreased the toxicity of cadmium in rats. In vitro the antitoxic activity of methanol extract of H. cordata $T_{HUNB}$ on NIH 3T3 fibroblasts was evaluated by the MTT {3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide} and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treated cells, $10^{-2}\;mg/ml$ Concentrations of H. cordata $T_{HUNB}$ extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that H. cordata $T_{HUNB}$ extract retains a potential antitoxic activity.

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Nerve Growth Factor Stimulates Glioblastoma Proliferation through Notch1 Receptor Signaling

  • Park, Jun Chul;Chang, In Bok;Ahn, Jun Hyong;Kim, Ji Hee;Song, Joon Ho;Moon, Seung Myung;Park, Young-Han
    • Journal of Korean Neurosurgical Society
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    • v.61 no.4
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    • pp.441-449
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    • 2018
  • Objective : Notch receptors are heterodimeric transmembrane proteins that regulate cell fate, such as differentiation, proliferation, and apoptosis. Dysregulated Notch pathway signaling has been observed in glioblastomas, as well as in other human malignancies. Nerve growth factor (NGF) is essential for cell growth and differentiation in the nervous system. Recent reports suggest that NGF stimulates glioblastoma proliferation. However, the relationship between NGF and Notch1 in glioblastomas remains unknown. Therefore, we investigated expression of Notch1 in a glioblastoma cell line (U87-MG), and examined the relationship between NGF and Notch1 signaling. Methods : We evaluated expression of Notch1 in human glioblastomas and normal brain tissues by immunohistochemical staining. The effect of NGF on glioblastoma cell line (U87-MG) was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. To evaluate the relationship between NGF and Notch1 signaling, Notch1 and Hes1 expression were evaluated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. To confirm the effects of NGF on Notch1 signaling, Notch1 and Hes1 small interfering RNAs (siRNAs) were used. Results : In immunohistochemistry, Notch1 expression was higher in glioblastoma than in normal brain tissue. MTT assay showed that NGF stimulates U87-MG cells in a dose-dependent manner. RT-PCR and Western blot analysis demonstrated that Notch1 and Hes1 expression were increased by NGF in a dose-dependent manner. After transfection with Notch1 and Hes1 siRNAs, there was no significant difference between controls and 100 nM $NGF-{\beta}$, which means that U87-MG cell proliferation was suppressed by Notch1 and Hes1 siRNAs. Conclusion : These results indicate that NGF stimulates glioblastoma cell proliferation via Notch1 signaling through Hes 1.

Antioxidant effect of Woogyuyeum against hydrogen peroxide-induced oxidative stress in Leydig cells (右歸飮이 hydrogen peroxide에 의해 유도된 Leydig cell에 미치는 항산화 효과 연구)

  • Kim, Soo Hyun;Kim, Do Rim;Chang, Mun Seog;Park, Seong Kyu
    • Herbal Formula Science
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    • v.23 no.1
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    • pp.111-119
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    • 2015
  • Objectives : The purpose of this study was to investigate the antioxidant activity of water extract of Woogyuyeum (WGY) in Leydig cells. Methods : We investigated the cytoprotective effect of WGY in cultured mouse Leydig cells by MTT assay. Leydig cells treated with WGY were incubated in the presence or absence of 50 μM hydrogen peroxide at 37℃ for 24 h. The protective effects of WGY against hydrogen peroxide-induced oxidative stress, lipid peroxide (LPO), superoxide dismutase (SOD), and catalase activity assays were performed in Leydig cells. Results : As a result, WGY showed no significant cytotoxicity in Leygdig cells. WGY showed cell viability as 103.65% in 5 μg/ml concentrations. The cytotoxicity induced by hydrogen peroxide in Leygdig cells, the antioxidant effects of WGY was increased in 1, 5, 50, 100 ug/ml concentraions. 100 μg/ml concentration of WGY showed maximum antioxidant effects. Treatment of cells with 100 μg/ml WGY significantly reduced the MDA concentration to 0.23 nmoles/mg protein. SOD activity was increased at 1, 100 μg/ml concentration of WGY and catalase activity was significantly increased at 50, 100 μg/ml concentrations of WGY, respectively. Conclusions : In conclusion, WGY has antioxidant activities against hydrogen peroxide-induced oxidative stress in Leydig cells.

Characteristic of neuroblastoma cell (SH-SY5Y) culture on the crystalline diamond film (다결정 다이아몬드 필름의 신경종양세포(SH-SY5Y) 배양 특성)

  • Nam, Hyo-Geun;Oh, Hong-Gi;Kim, Dae-Hoon;Kim, Min-Hye;Park, Hye-Bin;Jhee, Kwang-Hwan;Song, Kwang-Soup
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.12 no.4
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    • pp.10-15
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    • 2013
  • In order to fabricate high sensitive and stable biosensors, we require the material with superior biocompatibility and physical-chemical stability. Many kinds of biomaterials have been evaluated to apply for bioindustry. Recently, carbon based diamond thin films have been focal pointed as bio-applications and their possibility has been evaluated. Diamond thin film has many advantages for electrochemical and biological applications, such as wide potential window (3.0-3.5V), low background current and chemical-physical stability. In this work, we have cultured neuroblastoma cell (SH-SY5Y) on the crystalline diamond films. We use MTT assay to evaluate the characteristic of cell culture on the substrates. As a result, neuroblastoma cell was cultured on the crystalline diamond film as similar as cell culture dish.

Anti-oxidative Effect of Some Plant Extracts Against Nitric Oxide-induced Oxidative Stress on Neuronal Cell (Nitric oxide에 의해 산화적 스트레스를 받은 Neuronal cell에 항산화 효과를 가지는 수종 생약추출물의 검색)

  • Koo, Uk;Lee, Hak-Ju;Lee, Dong-Ho;Lee, Hyun-Jung;Ham, Ah-Rom;Cho, Eun-Young;Mar, Woong-Chon
    • Korean Journal of Pharmacognosy
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    • v.39 no.4
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    • pp.290-294
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    • 2008
  • The objective of this study is screening the anti-oxidative effects of several plant MeOH extracts against oxidative stress in Neuroblastoma cell. Oxidative stress has been implicated in the pathogenesis of many neurotoxicity, neurodegenerative disorders and cell death. This oxidative stress is generated by ROS (Reactive Oxygen Species) such as nitric oxide, nitrogen dioxide, peroxyl, superoxide ($O_2^-$), hydroxyl, alkoxyl. So, in the present study, we induced oxidative stress by treatment of sodium nitroprusside (2.5 mM) in human neuroblastoma SH-SY5Y cell which was treated samples before 24hr, and cell viability was measured by MTT reduction assay. Of those tested, the extracts of Paeonia japonica (roots), Eucommia ulmoides (炒)(barks), Paeonia japonica (曝乾)(roots), Phyllostachys bambusoides (stems), Polygala tenuifolia (去心, 炒)(roots), Paeonia japonica (roots), Polygala tenuifolia (roots), Machilus thunbergii (barks), Mallotus japonicus (leaves), Poria cocos (whole), Sophora flavescens (roots), Angelica tenuissima (roots), Angelica gigas (當歸尾)(roots) showed anti-oxidative effects[$EC_{50}$<15.20 ${\mu}g$/ml(Carnosine:Positive control)]in dose dependent manner.

The Effect of an Essential Oil Fragrance from Radix Angelica Sinesis on Differentiation and Proliferation of Neural Stem Cells of Rat (당귀(當歸) 향기액(香氣液)이 Rat의 뇌신경줄기세포의 분화(分化)와 증식(增殖)에 미치는 영향)

  • Park, Se-Hwan;Kang, Jae-Hyun;Jung, Young-Su;Kim, Geun-Woo;Koo, Byung-Soo
    • Journal of Oriental Neuropsychiatry
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    • v.20 no.4
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    • pp.63-77
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    • 2009
  • Objectives : In this study, an essential oil fragrance from Danggwi was administrated into the neural stem cell and the effect of the essential oil on the differentiation and proliferation of the neural stem cells were observed. Methods : The establishment of the neural stem cell was identified via Nestin, DAPI dye. An essential oil fragrance from Danggwi was administrated with a proved optimum level for the survival of the cell through MTT assay. Also, according to the analysis of Western blot, the essential oil fragrance from Danggwi promotes the phosphorylating of Akt, Erk, ERM protein. Results : MTT assay showed increased in GFAP. The result indicates that the differentiation to astrocyte is promoted. The phosphorylation levels of ERM, Erk and Akt were increased at 60 min after addition of 5 ug/ml of essential oil fragrance from Danggwi and sustained to 48 hours. These imply that essential oil fragrance from Danggwi may induce the survival and the proliferation of the differentiated cells. Conclusions : These results suggest that the essential oil fragrance from Danggwi can be effective for the in vivo study of degenerative neuronal disease using neural stem cell.

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Regenerative Effects of Houttuynia cordata Extracts against Cadmium-induced Cytotoxicity in NIH 3T3 fibroblasts Cultures (VII) (카드뮴 독성으로 손상된 NIH 3T3 섬유모세포에 대한 어성초 추출물의 재생효과(VII))

  • Lee Jeong-Ho;Oh Hyun-Ju;Jeong Seung-Il;Chun Hyun-Ja;Han Do-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.122-128
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    • 2001
  • This study was carried out to evaluate antitoxic effects of Houttuynia cordata extract on cadmium by colorimetric methods. The antitoxic activity ofc in NIH 3T3 fibroblasts was evaluated by 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. The concentration of 10-2 mg/ml of Houttuynia cordata extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were antitoxic and tend to regenerate. These results suggest that the ethyl acetate extract of Houttuynia cordata retains a potential antitoxic activity.

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Treatment of BG-1 Ovarian Cancer Cells Expressing Estrogen Receptors with Lambda-cyhalothrin and Cypermethrin Caused a Partial Estrogenicity Via an Estrogen Receptor-dependent Pathway

  • Kim, Cho-Won;Go, Ryeo-Eun;Choi, Kyung-Chul
    • Toxicological Research
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    • v.31 no.4
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    • pp.331-337
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    • 2015
  • Synthetic pyrethroids (SPs) are the most common pesticides which are recently used for indoor pest control. The widespread use of SPs has resulted in the increased exposure to wild animals and humans. Recently, some SPs are suspected as endocrine disrupting chemicals (EDCs) and have been assessed for their potential estrogenicity by adopting various analyzing assays. In this study, we examined the estrogenic effects of lambda-cyhalothrin (LC) and cypermethrin (CP), the most commonly used pesticides in Korea, using BG-1 ovarian cancer cells expressing estrogen receptors (ERs). To evaluate the estrogenic activities of two SPs, LC and CP, we employed MTT assay and reverse-transcription polymerase chain reaction (RT-PCR) in LC or CP treated BG-1 ovarian cancer cells. In MTT assay, LC ($10^{-6}M$) and CP ($10^{-5}M$) significantly induced the growth of BG-1 cancer cells. LC or CP-induced cell growth was antagonized by addition of ICI 182,720 ($10^{-8}M$), an ER antagonist, suggesting that this effect appears to be mediated by an ER-dependent manner. Moreover, RT-PCR results showed that transcriptional level of cyclin D1, a cell cycle-regulating gene, was significantly up-regulated by LC and CP, while these effects were reversed by co-treatment of ICI 182,780. However, p21, a cyclin D-ckd-4 inhibitor gene, was not altered by LC or CP. Moreover, $ER{\alpha}$ expression was not significantly changed by LC and CP, while down-regulated by E2. Finally, in xenografted mouse model transplanted with human BG-1 ovarian cancer cells, E2 significantly increased the tumor volume compare to a negative control, but LC did not. Taken together, these results suggest that LC and CP may possess estrogenic potentials by stimulating the growth of BG-1 ovarian cancer cells via partially ER signaling pathway associated with cell cycle as did E2, but this estrogenic effect was not found in in vivo mouse model.

Antitumor Effect and the Change of Chemosensitivity of Chitosan in Human Lung Cancer Cell Line (인체 폐암세포주에 대한 키토산의 항암효과와 항암제 감수성에 미치는 영향)

  • 노숙령
    • Journal of Nutrition and Health
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    • v.31 no.4
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    • pp.739-746
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    • 1998
  • This study was designed to investigated the antitumor effect and change chemosensitivity of chitosan in 2 kinds of humen lung cancer cell lines(NCI-H522, NCI-H596). To evaluate the antitumor effect and synergistic effectof chomosensitivity, MTT assay was used in vitro. then anticancer drugs used were 챤-platin , ectoposide, and adrimycin. The results of this study were as follows; Chitosan shwoed in antitumor effect on both NCI-H522 and NCI-H596. The lung cancer viability percent for NCI-H522 and NCL-H596 showed at the lowest levels of 5.31 and 5.33% when the concentration of chitosan was 25mg/$m\ell$ media and the exposure time of chitosan was 72 hours. ID50 value of chitosan on both NCI-H522 and NCI-H596 showed at the lowest levels of 14.07, 11.68 mg/$m\ell$ media when the exposure time of chitosan was 72 hours. the synergistic effect of chomosensitivity was better in NCI-H596 than in NCI0H522 . When the synergistic effect of chomosensitivity was shown according to the kind of the anticancer drugs, in case of NCI-H522 , in the concentration of 100$\mu\textrm{g}$/$m\ell$, ectoposide showed the highest synergistic effect of chomosensitivity and then was adrimycin In case of NCI-H596, in the concentration of 100$\mu\textrm{g}$/$m\ell$,, the order of the synergistic effect of chomosensitivity was ectoposide>adrimycin>cis-platin and in the concentration of 10$\mu\textrm{g}$/$m\ell$, ectoposide>cis-platin >adrimycin. It is concluded that chitosan is an active antitumor agent and is increased chomosensitivity though there is difference according to the kind and the concentration of anticancer drugs. But to be sued to lung cancer theraphy, further studies on toxicity, the mechanism of action, animal experiment are wanted.

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Ellagic Acid Exerts Anti-proliferation Effects via Modulation of Tgf-Β/Smad3 Signaling in MCF-7 Breast Cancer Cells

  • Zhang, Tao;Chen, Hong-Sheng;Wang, Li-Feng;Bai, Ming-Han;Wang, Yi-Chong;Jiang, Xiao-Feng;Liu, Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.273-276
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    • 2014
  • Ellagic acid has been shown to inhibit tumor cell growth. However, the underlying molecular mechanisms remain elusive. In this study, our aim was to investigate whether ellagic acid inhibits the proliferation of MCF-7 human breast cancer cells via regulation of the TGF-${\beta}$/Smad3 signaling pathway. MCF-7 breast cancer cells were transfected with pEGFP-C3 or pEGFP-C3/Smad3 plasmids, and treated with ellagic acid alone or in combination with SIS3, a specific inhibitor of Smad3 phosphorylation. Cell proliferation was assessed by MTT assay and the cell cycle was detected by flow cytometry. Moreover, gene expression was detected by RT-PCR, real-time PCR and Western blot analysis. The MTT assay showed that SIS3 attenuated the inhibitory activity of ellagic acid on the proliferation of MCF-7 cells. Flow cytometry revealed that ellagic acid induced G0/G1 cell cycle arrest which was mitigated by SIS3. Moreover, SIS3 reversed the effects of ellagic acid on the expression of downstream targets of the TGF-${\beta}$/Smad3 pathway. In conclusion, ellagic acid leads to decreased phosphorylation of RB proteins mainly through modulation of the TGF-${\beta}$/Smad3 pathway, and thereby inhibits the proliferation of MCF-7 breast cancer cells.