• Title/Summary/Keyword: MTS Assay

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Induction of Apoptosis Signaling by a Glycoprotein of Capsosiphon fulvescens in AGS Cell (매생이 (Capsosiphon fulvescens) 당단백질에 의한 인간 위암세포 사멸기전)

  • Kim, Young-Min;Kim, In-Hye;Nam, Taek-Jeong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.3
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    • pp.216-224
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    • 2011
  • Capsosiphon fulvescens is well-known green sea algae that, in recent years, has been proposed as a potential anticancer drug. In this study, we found that C. fulvescens glycoprotein (Cf-GP) had pro-apoptotic effects on human gastric carcinoma cells. By SDS-PAGE, we confirmed that C. fulvescens extract contained a glycoprotein. Using H33342 staining, we found that the Cf-GP caused cell death in a does-dependent manner, while an MTS assay showed decreased cellular viability due to induction of apoptosis. To determine the effect of Cf-GP on apoptosis-related cellular events, cells were treated with Cf-GP and the expression of several apoptosis-related protein was determined by Western blotting. Our results indicate that Cf-GP activated both a caspase cascade and PARP, which is a substrate of caspase-3, caspase-8 and the Bcl-2 family proteins. In addition, we assessed caspase-3, and -8 activation and annexin V staining. Our results revealed a cell cycle arrest, itself leading to an increased percentage of sub-G1 cells. Our findings indicate that Cf-GP may be a source of bio-functional material with therapeutic effects on human gastrointestinal cancer.

Anti-inflammatory Effect of Mugi-hwan Water Extract in RAW 264.7 Cells (무기환(戊己丸)의 RAW 264.7 세포에 대한 항염증작용 연구)

  • Kim, Ilhyun;Choi, Chonghwan;Lee, Sewon;Song, Yungsun
    • Journal of Korean Medicine Rehabilitation
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    • v.23 no.3
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    • pp.27-35
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    • 2013
  • Objectives The aim of this study was to investigate anti-inflammatory activity of Mugi-hwan (MGH) Water Extract. Methods Cells were treated with 2 ug/ml of LPS 1 hour prior to the addition of MGH. Cell viability was measured by MTS assay. The production of NO was determined by reacting cultured medium with Griess reagent. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. The content of level of cytokines ($PGE_2$, IL-6, in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. Results MGH inhibited the production of NO, $PGE_2$, IL-6 as well as the expressions of iNOS, COX-2 in the murine macrophage, RAW 264.7 cells. MGH also had suppression effects of LPS induced MAPKs activation. Conclusions These results suggest that MGH has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes.

Effects of Kochiae Fructus Extracts on the Expression of Angiogeneis and Inflammation Related Proteins (지부자(地膚子)의 신생혈관 및 염증매개 단백질 발현에 미치는 영향)

  • Na, Sang-Hyuk;Shin, Yong-Cheol;Ko, Seong-Gyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.3
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    • pp.557-562
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    • 2006
  • Hypoxia induced angiogenesis and inflammation are essential processes for metastasis and progress of solid tumors. We examined the anti-angiogenic and inflammation related activity of Kochiae Fructus (KF) extract. To investigate the roles of the KF extract, we performed MTS assay, western blots using HaCaT cells and $HepG_2$ cells. The results are as follows. The protein level of $HIF-1{\alpha}$ was reduced when induced by $CoCl_2$ in $HepG_2$ cells treated with KF extract and induced by IGF-11 in HaCaT cells treated with KF extract. KF extract reduced the mRNA level of VEGF in HaCaT cells and KF extract reduced the protein level of iNOS in HaCaT cells. These results suggest that KF extract contributes to the anti-angiogenic and anti-inflammatory activities and also we could assume that KF extract act as antioxidant or anti-inflammatory agents via reduction of $HIF-1{\alpha}$.

Influence of DBD Plasma Exposure on Normal and Cancer Cells Activity

  • Panngom, Kamonporn;Baik, Ku-Youn;Ryu, Young-Huo;Choi, Eun-Ha
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.172-172
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    • 2012
  • Non-thermal plasma has attracted medical researchers, since they showed higher apoptosis rate in cancer cells than normal cells. However, it is hard to conclude general cancer cell specific effect because comparison between normal and cancer cell activities after plasma treatment have not been reported yet. This research proposes a comparison of Dielectric Barrier Discharge (DBD) plasma effect on three normal cells lines and three cancer cells lines. We measured cell number, mitochondria activity (MTS assay) and amount of hydrogen peroxide (H2O2) for three days. The results show that the number of cancer cells decreased more than normal cells following of exposure time. On the other hand, mitochondria activities and amounts of H2O2 increased following of exposure time. In addition, we found that DBD plasma exposure on cell suspension in media and media only illustrated no difference in mitochondria activity, H2O2 quantity, and cell number. Thus, we can confirm higher apoptosis rate in cancer cells which is related to the reactive oxygen species (ROS) generated by DBD plasma. The related molecular mechanisms were investigated further.

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Anti-Inflammatory Effects of Rice Bran Ethanol Extract in Murine Macrophage RAW 264.7 Cells (미강에탄올추출물의 RAW264.7 세포에서 항염증효과)

  • Park, Jeong-Suk;Kim, Mi-Hye
    • YAKHAK HOEJI
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    • v.55 no.6
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    • pp.456-461
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    • 2011
  • The aim of the present study is to investigate the anti-inflammatory effect of a Rice Bran Ethanol Extract (RBE). Inflammation, such as a bacterial infection in vivo metabolites, such as external stimuli or internal stimuli to the defense mechanisms of the biological tissue a variety of intracellular regulatory factors deulin inflammatory TNF-${\alpha}$, IL-$1{\beta}$, IL-6, IL-8, such as proinflammatory cytokines, prostagrandin, lysosomal enzyme, free radicals are involved in a variety of mediators. The present study was designed to determine the effect of the RBE on pro-inflammatory factors such as NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of RBE, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, the RBE reduced NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 production without cytotoxicity. Our results suggest that the RBE may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful as an anti-inflammatory material.

Anti-oxidative and Anti-inflammatory Effect of 7 Herbal Extracts and Methods of Herbal Formula Compositioning (7종 한약재 추출물의 항산화 및 항염증 효과와 이를 이용한 방제 구성에 대한 고찰)

  • Park, JeongHoo;Park, JooYeon;Park, Sundong
    • Herbal Formula Science
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    • v.22 no.2
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    • pp.87-103
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    • 2014
  • Objectives : Herbal formulas are complicated to analyze and difficult to compose because of its mixed, complex features. So we discussed about how to compose herbal formulas effective and efficient by analyzing various effectiveness of each herbal extracts. Methods : To evaluate the effectiveness of herbal formula, anti-oxidative and anti-inflammatory effectiveness were mathematically analyzed. DPPH, superoxide anions, Nitric Oxide scavenging activity was measured to evaluate the effectiveness of 7 herbal extracts. And next, cytotoxic activity of extracts on RAW 264.7 cells were measured using MTS assay. To asses anti-inflammatory effect, nitric oxide and $PGE_2$ production were measured. Based on these anti-oxidative and anti-inflammatory experiment result, mathematical analyzation were carried out with constOptm function, and determined efficacy-maximizing ratio. Results : This mathematical analysis based formula showed significantly outstanding effectiveness than other formulae. And estimated tendency of anti-inflammatory effectiveness was matched with real effectiveness. Conclusions : So, mathematical analysis can be available to evaluate and estimate the effectiveness of herbal formulae.

Anti-inflammatory Effect of Imyosan Extract is more potent than that of its Component Herb Extracts in Murine Macrophages (마우스 대식세포인 RAW 264.7에 대한 이묘산(二妙散)의 메탄올 추출물의 항염증 효과)

  • Yun, Hyun-Jeong;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.163-173
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    • 2008
  • Objectives : Imyosan (IMS), a drug preparation comprised of Phellodendri Cortex (PC) and Atractylodis Rhizoma (AR), is commonly used as a traditional herbal medicine in Korea and China for the treatment of various inflammatory diseases. However, little is known about the effect of IMS and its component herbs on inflammatory mediators in RAW 264.7 cells. Therefore, in this study, methanol extracts of IMS and its component herbs were examined to determine if they inhibited inflammatory effects in RAW 264.7 cells. Methods : Cytotoxic activity of IJHT and its components on RAW 264.7 cells was using 5-(3-carboxymethoxyphenyl)-2H-tetrazolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were detected by western blot. Results : Methanol extract of IMS and its component herbs were significantly reduced iNOS and COX-2 expression as well as NO, PGE2, $IL-1{\beta}$ and IL-6 production in RAW 264.7 cells. Conclusions : The results of this study indicated that the anti-inflammatory effect of Imyosan extract is more potent than that of extracts of its component herbs in macrophages.

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Scutellaria baicalensis ethanol extracts inhibit IGF-II-induced HIF-1 ${\alpha}$ and VEGF expressions in HaCaT cells. (황금 에탄올 추출물이 IGF-II로 유도된 $HIF-1{\alpha}$와 VEGE 발현 억제에 미치는 영향)

  • Byun, Hak-Sung;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.20 no.1 s.32
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    • pp.27-37
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    • 2007
  • Background and Objective : Psoriasis is a chronic skin disease characterized by angiogenesis. It has been reported that growth factor as vascular endothelial growth factor (VEGF) and insulin-like growth factor (IGF)-II are overexpressed in psoriatic epidermis. This stydy was carried out for whether SB extracts have an anti-angiogenic effect for angiogenic factor. Method : To investigate the inhibitory effect of VEGF expression by the SB extracts, we performed MTS assay, western blots using HaCaT cells. HaCaT cells were pretreated with SB extracts for 1 hour followed by treatment with IGF-II. Result : SB extracts significantly reduced IGF-II induced HIF-1 ${\alpha}$ protein level via p53 and MAPK pathway in HaCaT cells. Also, SB extracts inhibited IGF-II induced VEGF mRNA and protein expression levels in the HaCaT keratinocytes. Conclusion : These results suggest that inhibition of HIF-1 ${\alpha}$ and VEGF expressions by SB extracts contributes to the anti-angiogenic effects.

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Anticancer and Signaling Mechanisms of Biologically Active Substances from Orostachys japonicus through Arrest of Cell cycle in Human Melanoma Cells (인체 흑색종 세포에 대한 와송 추출물의 세포주기 억제를 통한 항암효과와 기전 연구)

  • Ryu, Deok-Hyun;Ryu, Deok-Seon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.32 no.4
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    • pp.1-12
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    • 2019
  • Objectives : The purpose of this study was to identify the anticancer effect of biological substances of ethylacetate(EtOAc) fraction from Orostachys japonicus(OJEF), their effect on human melanoma A375 cells and the related molecular mechanisms. Methods : The MTS assay was used to confirm the inhibition of cancer cell proliferation in A375 cells. And the $MUSE^{TM}$ analyzer was used to determine the ability of OJEF to induce cell cycle arrest. Western blotting was used to determine the changes in protein expression in A375 cells after treatment with OJEF. Results : OJEF showed cytotoxicity to A375 cells. And cell cycle arrest occurred in G1 phase and G2/M phase owing to inhibition of CDK1, cyclin B1, CDK4, and cyclin D, which are related to cell cycle regulation and cell division control. Conclusion : OJEF is effective in regulating cell cycle of human melanoma cells and thus can be a good theraputic agent to treat patients with melanoma.

Effects of Sandalwood Essential Oil on the iNOS Expression and Proinflammatory Cytokine Production (Sandalwood Essential Oil의 iNOS 발현과 염증성사이토카인의 생성에 미치는 영향)

  • Park, Jeoung-Suk;Jung, Sook-Heui
    • YAKHAK HOEJI
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    • v.57 no.1
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    • pp.70-75
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    • 2013
  • The present study was designed to determine the effect of the Sandalwood Essential Oil (Santalum album) on pro-inflammatory factors such as NO, iNOS expression and IL-$1{\beta}$, IL-6, TNF-${\alpha}$ in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of Sandalwood Essential Oil, amount of NO was measured using the NO detection kit and the iNOS expression was measured by western blot analysis and reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, Sandalwood Essential Oil reduced NO, iNOS expression and IL-$1{\beta}$, IL-6, TNF-${\alpha}$ production without cytotoxicity. Our results suggest that the Sandalwood Essential Oil may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful as an anti-inflammatory oil.