• Title/Summary/Keyword: 5-diphenyl-2H-tetrazolium bromide

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Moutan Cortex Extract Inhibits Amyloid ${\beta}$ Protein (25-35)-induced Neurotoxicity in Cultured Rat Cortical Neurons (Amyloid ${\beta}$ 2 Protein (25-35) 유도 배양신경세포 독성에 대한 목단피의 억제효과)

  • Kim, Joo-Youn;Ju, Hyun-Soo;Ban, Ju-Yeon;Song, Kyung-Sik;Seong, Yeon-Hee
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.6
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    • pp.409-415
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    • 2008
  • Moutan cortex, the root bark of Paeonia suffruticosa Andrews (Paeoniaceae), has pharmacological effects such as anti-inflammatory, antiallergic, analgesic and antioxidant activities. We investigated a methanol extract of Moutan cortex for neuroprotective effects on neurotoxicity induced by amyloid ${\beta}$ protein ($A{\beta}$) (25-35) in cultured rat cortical neurons. Exposure of cultured cortical neurons to $10\;{\mu}M\;A{\beta}$ (25-35) for 24 h induced neuronal apoptotic death. Moutan cortex inhibited $10\;{\mu}M\;A{\beta}$ (25-35)-induced neuronal cell death at 30 and $50\;{\mu}g/m{\ell}$, which was measured by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. Moutan cortex inhibited $10\;{\mu}M\;A{\beta}$ (25-35)-induced elevation of intracellular calcium concentration ($[Ca^{2+}]_i$), and generation of reactive oxygen species (ROS) which were measured by fluorescent dyes. Moutan cortex also inhibited glutamate release into medium induced by $10\;{\mu}M\;A{\beta}$ (25-35), which was measured by HPLC. These results suggest that Moutan cortex prevents $A{\beta}$ (25-35)-induced neuronal cell damage by interfering with the increase of $[Ca^{2+}]_i$, and then inhibiting glutamate release and ROS generation. Moutan cortex may have a therapeutic role in preventing the progression of Alzheimer's disease.

Effect of STAT3 on Lysophosphatidic Acid-Induced Oral Cancer Cell Invasion

  • Song, Zi Hae;Cho, Kyung Hwa;Kim, Jin Young;Lee, Hoi Young
    • Journal of dental hygiene science
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    • v.19 no.2
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    • pp.141-146
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    • 2019
  • Background: Oral cancer has a high incidence worldwide and has been closely associated with smoking, alcohol, and infection by the human papillomavirus. Metastasis is highly important for oral cancer survival. Lysophosphatidic acid (LPA) is a bioactive lipid mediator that promotes various cellular processes, including cell survival, proliferation, metastasis, and invasion. Signal transducer and activator of transcription (STATs) are transcription factors that mediate gene expression. Among the seven types of STATs in mammals, STAT3 is involved in invasion and metastasis of numerous tumors. However, little is known about the role of STAT3 in oral tumor invasion. In the present study, we hypothesized that STAT3 mediates LPA-induced oral cancer invasion. Methods: Immunoblotting was performed to analyze LPA-induced STAT3 activation. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was performed to assess the survival rates of YD-10B cells. STAT3 levels in LPA-treated oral tumor cells were evaluated by performing in vitro invasion assay. Results: To the best of our knowledge, this is the first study to demonstrate that LPA enhances STAT3 phosphorylation in oral cancer. In addition, treatment with WP1066, a selective inhibitor of STAT3, at a concentration that does not cause severe reduction in cell viability, significantly attenuated LPA-induced YD-10B cancer cell invasion. Conclusion: The results suggested that LPA induces oral tumor cells with greater invasive potential via STAT3 activation. Our findings provided important insights into the mechanisms underlying mouth neoplasms.

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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Investigation of ${\alpha}$-Glucosidase Inhibitory Activity of Ethanolic Extracts from 19 Species of Marine Macroalgae in Korea

  • Jeong, So-Young;Qian, Zhong-Ji;Jin, Yeong-Jun;Kim, Gi-Ok;Yun, Pil-Yong;Cho, Tae-Oh
    • Natural Product Sciences
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    • v.18 no.2
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    • pp.130-136
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    • 2012
  • In the present work, we have collected 19 species of macroalgae (9 Phaeophta and 10 Rhodophyta) f rom all around of Korea: Dictyopteris divaricata, D. prolifera, Myelophycus cavus, Papenfussiella kuromo, Petalonia zosterifolia, Petrospongium rugosum, Rugulopteryx okamurae, Sargassum fulvellum, S. muticum, Callophyllis japonica, Gloiopeltis tenax, Gracilaria longissima, Gracilaria vermiculophylla, Grateloupia asiatica, Grateloupia lanceolata, Grateloupia sparsa, Grateloupia turuturu, Grateloupia sp, and Polyopes affinis. The macroalgal species were extracted by 70% ethanol (EtOH) for 24 h and evaluated its inhibitory effects on ${\alpha}$-glucosidase. Among ethanol extracts, Myelophycus cavus showed the most effectively inhibitory activity ($IC_{50}$, 2.17 ${\mu}g/ml$) against ${\alpha}$-glucosidase, followed by Sargassum fulvellum (<$IC_{50}$, 8.13 ${\mu}g/ml$), Dictyopteris prolifera ($IC_{50}$, 16.66 ${\mu}g/ml$), Rugulopteryx okamurae ($IC_{50}$, 50.63 ${\mu}g/ml$), and Petrospongium rugosum ($IC_{50}$, 101.62 ${\mu}g/ml$). Furthermore, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay showed no cytotoxicity on mouse pre-adipocytes cell line (3T3-L1). These results suggest that some edible macroalgae merit further evaluation for clinical usefulness as anti-diabetic functional foods.

Effects of Kuseonwangdogo on the Proliferation of Preadipocyte 3T3-L1 Cells, the Anti-Complementary and the Cytotoxic Effects (구선왕도고가 전지방세포(前脂肪細胞) 3T3-L1의 증식(增殖), 항보체활성(抗補體活性) 및 세포독성(細胞毒性)에 미치는 영향)

  • Choi, Young-Heu;Kim, Ho-Kyoung;Ko, Byoung-Seob;Ju, Young-Sung
    • The Journal of Korean Medicine
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    • v.20 no.3 s.39
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    • pp.105-114
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    • 1999
  • To investigate the anti-complementary and cytotoxic effects of oriental prescription, Kuseonwangdogo, on the proliferation of preadipocyte 3T3- L1 cells, we examined biological effects of Kuseonwangdogo. The results obtained were as follows. 1. After 14 days, the body weight of rats treated with Kuseonwangdogo decreased more than that in the control group (p<0.05). However, the weights of liver, spleen and kidney were unchanged. In serum biochemical test, we examined the level of glucose (GLU) and glutamic pyruvic transaminase (GPT). The levels of GOT and CHOL in serum were decreased remarkably by the administration of Kuseonwangdogo (p<0.05). The haematological examination of the tested group showed significant increment of white blood cells (WBC), hemoglobin concentration (HGB), mean corpuscular hemoglobin (MCH) and monocyte (MO). 2. The effect of Kuseonwangdogo on the proliferation of 3T3-L1 cells was tested by the sulforhodamin B(SRB) assay. The high concentration ($100{\mu}l\;and\;200{\mu}l$) of extracts inhibited the proliferation of 3T3- L1 cells. The p-value was <0.01, respectively. 3. The extract of Kuseonwangdogo showed a potent anti -complementary activity. It was suggested that the active principle may be a kind of polysaccharide molecule. 4. The cytotoxic effects of Kuseonwang dogo and its composing herbs in human liver cells (WRL68) and monkey kidney cells (Vero) were examined by the SRB and 3- (4,5- Dimethylthiazol-2-yl) -2,5 diphenyl-2H- tetrazolium bromide (MTT) assay. Cytotoxic effects were not observed.

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Ginsenoside Rg3 reduces the adhesion, invasion, and intracellular survival of Salmonella enterica serovar Typhimurium

  • Mechesso, Abraham F.;Quah, Yixian;Park, Seung-Chun
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.75-85
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    • 2021
  • Background: Invasive infections due to foodborne pathogens, including Salmonella enterica serovar Typhimurium, are prevalent and life-threatening. This study aimed to evaluate the effects of ginsenoside Rg3 (Rg3) on the adhesion, invasion, and intracellular survival of S. Typhimurium. Methods: The impacts of Rg3 on bacterial growth and host cell viability were determined using the time kill and the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays, respectively. Gentamicin assay and confocal microscopic examination were undertaken to determine the effects of Rg3 on the adhesive and invasive abilities of S. Typhimurium to Caco-2 and RAW 264.7 cells. Quantitative reverse transcription polymerase chain reaction was performed to assess the expression of genes correlated with the adhesion, invasion, and virulence of S. Typhimurium. Results: Subinhibitory concentrations of Rg3 significantly reduced (p < 0.05) the adhesion, invasion, and intracellular survival of S. Typhimurium. Rg3 considerably reduced (p < 0.05) the bacterial motility as well as the release of nitrite from infected macrophages in a concentration-dependent manner. The expression of genes related to the adhesion, invasion, quorum sensing, and virulence of S. Typhimurium including cheY, hilA, OmpD, PrgK, rsgE, SdiA, and SipB was significantly reduced after Rg3 treatment. Besides, the compound downregulated rac-1 and Cdc-42 that are essential for actin remodeling and membrane ruffling, thereby facilitating Salmonella entry into host cells. This report is the first to describe the effects of Rg3 on "trigger" entry mechanism and intracellular survival S. Typhimurium. Conclusion: Rg3 could be considered as a supplement agent to prevent S. Typhimurium infection.

Prognostic Value of T Cell Immunoglobulin Mucin-3 in Prostate Cancer

  • Piao, Yong-Rui;Piao, Long-Zhen;Zhu, Lian-Hua;Jin, Zhe-Hu;Dong, Xiu-Zhe
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3897-3901
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    • 2013
  • Background: Optimal treatment for prostate cancer remains a challenge worldwide. Recently, T cell immunoglobulin mucin-3 (TIM-3) has been implicated in tumor biology but its contribution prostate cancer remains unclear. The aim of this study was to investigate the role of TIM-3 as a prognostic marker in patients with prostate cancer. Methods: TIM-3 protein expression was determined by immunohistochemistry and Western blotting in 137 prostate cancer tumor samples and paired adjacent benign tissue. We also performed cell proliferation assays using 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl- 2H tetrazolium bromide (MTT) and cell invasion assays. The effects of small interfering RNA (siRNA)-mediated knockdown of TIM-3 (TIM-3 siRNA) in two human prostate cancer cell lines were also evaluated. Results: TIM-3 expression was higher in prostate cancer tissue than in the adjacent benign tissue (P<0.001). High TIM-3 expression was an independent predictor of both recurrence-free survival and progression-free survival. TIM-3 protein was expressed in both prostate cancer cell lines and knockdown suppressed their proliferation and invasion capacity. Conclusions: TIM-3 expression is associated with a poor prognosis in prostate cancer. Taken together, our resutlts indicate that TIM-3 is a potential prognostic marker in prostate cancer.

Neuronal Cell Protective Effects of Methanol Extract from Cheonggukjang Using in vitro System (In vitro system에서 청국장 메탄올 추출물의 신경세포 보호효과)

  • Jeong, Chang-Ho;Kwak, Ji-Hyun;Kim, Ji-Hye;Choi, Gwi-Nam;Jeong, Hee-Rok;Heo, Ho-Jin
    • Korean Journal of Food Science and Technology
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    • v.42 no.6
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    • pp.768-772
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    • 2010
  • In this study, the neuronal cell protective effects of methanol extract from cheonggukjang were evaluated. The proximate composition and total phenolics of the methanol extract were 40.95% crude protein, 22.49% crude fat, 15.99% nitrogen free extract, 7.91% moisture, 6.74% crude ash, 5.92% crude fiber, and 28.43 mg/g of total phenolics. Intracellular ROS accumulation resulting from $H_2O_2$ treatment of PC12 cells was significantly reduced when methanol extract was present in the media compared to PC12 cells treated with $H_2O_2$ only. In a cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium-bromide (MTT), the methanol extract showed protective effects against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase (LDH) release into the medium was also inhibited. Furthermore, the inhibitory effect of the methanol extract against acetylcholinesterase was dose-dependent.

Endothelial Cell Products as a Key Player in Hypoxia-Induced Nerve Cell Injury after Stroke

  • Cho, Chul-Min;Ha, Se-Un;Bae, Hae-Rahn;Huh, Jae-Taeck
    • Journal of Korean Neurosurgical Society
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    • v.40 no.2
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    • pp.103-109
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    • 2006
  • Objective : Activated endothelial cells mediate the cascade of reactions in response to hypoxia for adaptation to the stress. It has been suggested that hypoxia, by itself, without reperfusion, can activate the endothelial cells and initiate complex responses. In this study, we investigated whether hypoxia-induced endothelial products alter the endothelial permeability and have a direct cytotoxic effect on nerve cells. Methods : Hypoxic condition of primary human umbilical vein endothelial cells[HUVEC] was induced by $CoCl_2$ treatment in culture medium. Cell growth was evaluated by 3,4,5-dimethyl thiazole-3,5-diphenyl tetrazolium bromide [MTT] assay Hypoxia-induced products [$IL-1{\beta},\;TGF-{\beta}1,\;IFN-{\gamma},\;TNF-{\alpha}$, IL-10, IL-6, IL-8, MCP-l and VEGF] were assessed by enzyme-linked immunosorbent assay. Endothelial permeability was evaluated by Western blotting. Results : Prolonged hypoxia caused endothelial cells to secrete IL -6, IL -8, MCP-1 and VEGF. However, the levels of IL -1, IL -10, $TNF-{\alpha},\;TGF-{\beta},\;IFN-{\gamma}$ and nitric oxide remained unchanged over 48 h hypoxia. Hypoxic exposure to endothelial cells induced the time-dependent down regulation of the expression of cadherin and catenin protein. The conditioned medium taken from hypoxic HUVECs had the cytotoxic effect selectively on neuroblastoma cells, but not on astroglioma cells. Conclusion : These results suggest the possibility that endothelial cell derived cytokines or other secreted products with the increased endothelial permeability might directly contribute to nerve cell injury followed by hypoxia.

Stability of main components and physiological activities of bee venom treated with pH (산도에 따른 봉독의 성분 및 생리활성에 대한 안정성)

  • Cho, Miran;Han, Sangmi;Kim, Jungmin;Yeo, Joohong;Hong, InPhyo;Woo, Soonok;Lee, Kwanggill
    • Journal of Sericultural and Entomological Science
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    • v.52 no.1
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    • pp.6-9
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    • 2014
  • This study was for the investigation of the stability of purified bee venom (PBV) during the treatment in the pH range from pH2 to pH9 for 24 hours, respectively. Changes of components and physiological functionalities in PBV were by evaluated silver staining, and melittin contents were measured by liquid chromatography. The antimicrobial activity against bacteria by minimum inhibitory concentration (MIC) and effect of the cell regeneration were measured by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl tetrazolium bromide (MTT) assay using human dermal fibroblast (HDF) cell. The main proteins such as melittin and phospholipase $A_2$ showed no characteristic changes. The antimicrobial activity and effect of cell regeneration showed no difference from pH2 to pH9. From this study, we suggest that components and physiological functionalities of PBV against treated pH were kept stability at from pH2 to pH9.