• Title/Summary/Keyword: in vitro suppression

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Antimetastatic Effects of Capsaicin in Murine B16 Melanoma Cell Lines

  • Hwang Myung Sil;Yum Young Na;Kang Ho Il;Kim Ok Hee
    • Environmental Mutagens and Carcinogens
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    • v.25 no.1
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    • pp.1-5
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    • 2005
  • The present work was undertaken to evaluate the antimetastatic potential of capsaicin (8­methyl-N-vanillyl-6-nonenamide) by measuring its effects on matrix metalloproteinase activity, cell invasion and lung metastasis. Significant inhibition of matrix metalloproteinase-2 activity by capsaicin (100 $\mu$M) was detected by gelatin zymography. In vitro invasion assay showed capsaicin (50, 100 $\mu$M) reduced tumor cell invasion ($28-40\%$). Capsaicin (i.p., 2.5 mg/kg) inhibited development of lung colonization ($58\%$). These results suggest that capsaicin prevents metastasis in part through suppression of invasion of B16F10 melanoma cells by inhibiting matrix metalloproteinase-2 responsible for degradation of extracellular matrix.

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Epigallocatechin Gallate inhibits Prostagladins Generation by Suppression of cPLA2 Activity on Arachidonic Acid Metabolism in LPS-Stimulated RAW264.7 Cells

  • Son, Dong-Ju;Akiba, Satoshi;Sato, Takashi;Park, Young-Hyun;Yun, Yeo-Pyo
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.260.1-260.1
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    • 2002
  • Green tea contains several antioxidants including polyphenols of the catechin. which have been shown to act in vitro and in vivo as anti-inflammatory. anti-viral and anti-tumor drugs. Prostaglandins (PGs) are a family of intercellular and intracellular messengers derived from arachidonic acid(AA) by phospholipase(PL) and cyclooxygenase(COX). These mediators exert a wide range of effects on processes such as smooth muscle tone. vascular permeability, cellular proliferation. and inflammatory/immune function. (omitted)

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Anti-inflammatory Effect of Ribes fasciculatum in IFN-${\gamma}$/LPS-stimulated Mouse Peritoneal Macrophage

  • Kim, Jin Kyu;Kim, Ha Na;Kang, Chung Sik;Seo, Je Han;Seo, Hyun Won;Im, Jun Sang;Kim, Bong Seok;Cha, Dong Seok;Kwon, Jin;Oh, Chan Ho;Ma, Sang Yong;Nam, Jung Il;Jeon, Hoon
    • Natural Product Sciences
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    • v.20 no.2
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    • pp.113-118
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    • 2014
  • Ribes fasciculatum which belongs to Saxifragaceae has been widely used as a traditional medicine for the treatment of symptoms associated with lacquer poison. However, pharmacological studies on the R. fasciculatum are extremely limited until now. Thus, in this study, we evaluated the possible anti-inflammatory effects of ethyl acetate fraction of R. fasciculatum (ERF) using IFN-${\gamma}$/LPS-stimulated peritoneal macrophage model. We investigated the change in nitrite level in the absence or presence of ERF after LPS stimulation, and we found that ERF effectively attenuates the NO production in a dose dependent manner without notable toxicity. To determine the mechanism of the inhibitory action of ERF on NO production, we performed iNOS enzyme activity assay and Western blotting. Here we showed that both of iNOS enzyme activities and iNOS expressions were significantly down-regulated by ERF, indicating that these dual activities of ERF are responsible for ERF-mediated NO suppression. In addition, ERF inhibitied the expression of cyclooxygenase-2 (COX-2), an another key enzyme in inflammation through suppression of NF-${\kappa}B$ activation. We also tested anti-inflammatory properties of ERF not only in vitro, but in vivo using trypsin-induced paw edema model in mice. Our results revealed that the increased paw volume in response to trypsin injection was recovered by ERF supplement dose dependently.

PRODUCTION OF TRANSFORMING GROWTH FACTOR-${\beta}_1$ IN HUMAN FIBROBLASTS INDUCED WITH BACTERIAL TOXINS (세균 독소를 작용시킨 섬유아 세포에서 Transforming Growth Factor-${\beta}_1$의 생성)

  • Lee, Seong-Geun;Kim, Kwang-Hyuk
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.4
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    • pp.345-354
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    • 2000
  • TGF-${\beta}_1$ is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to wound healing. The objective of this study is to investigate production of TGF-${\beta}_1$ in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of TGF-${\beta}_1$ which may be responsible for wound healing. The fibroblasts were originated from facial dermis and hypertrophic scar in 26 year-old male patient. In the presence of LPS($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$), SEB($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$) respectively, cells($5{\times}10^3ml$) were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, cells($2.5{\times}10^5ml$) were cultivated in EMEM with LPS(0.01, 0.1 and $1.0{\mu}g$), SEB(0.01, 0.1 and $1.0{\mu}g$) respectively and LPS($0.1{\mu}g$) and SEB($0.1{\mu}g$) in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and TGF-${\beta}_1$ was assayed in duplicate. The results were as follows. 1. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of TGF-${\beta}_1$ was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of TGF-${\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. 2. In hypertrophic scar fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation did not occur at 1 day after incubation, compared with the control. In SEB and LPS exposure in combination, the production of TGF-${\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. However, the production of TGF-${\beta}_1$ did not occur in SEB and LPS exposure respectively. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of TGF-${\beta}_1$ very significantly and both fibroblasts have different phenotype each other in this regard. This data suggest that the significant production of TGF-${\beta}_1$ may develope abnormal wound healing associated with tissue fibroproliferative disorder, such as hypertrophic scar and keloid formation.

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Aspartyl-tRNA Synthetase from Acidithiobacillus ferrooxidans Aspartylates Both tRNA$^{Asp}$ and tRNA$^{Asn}$

  • Keem, Joo-Oak;Choi, Soon-Yong;Koh, Suk-Hoon;Hyun, Sung-Hee;Min, Bok-Kee
    • Biomedical Science Letters
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    • v.13 no.2
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    • pp.105-110
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    • 2007
  • Aspartyl-tRNA synthetase (AspRS) exists in two different forms with respect to tRNA recognition. The discriminating enzyme (D-AspRS) recognizes only tRNA$^{Asp}$, while the non-discriminating one (ND-AspRS) also recognizes tRNA$^{Asn}$ and therefore forms both Asp-tRNA$^{Asn}$ and Asp-tRNA$^{Asp}$. Plus primary sequence distinguishes two general groups of AspRS. There is a predominantly bacterial-type, larger AspRS (about 580 aa) in addition to a shorter archaeal/eukaryotic type (about 430 aa). In vivo data made clear that discriminating and non-discriminating enzymes exist in both groups. The determinants in the protein sequence responsible for tRNA discrimination are not hewn. The AspRS from Acidithiobacillus ferrooxidans might be suggested ND-AspRS fur missing of AsnRS in genomic sequencing data. Therefore, we analyzed the AspRS from A. ferrooxidans with in vitro aminoacylation assay with E. coli unfractionated tRNA, in vivo missense suppression assay with tipA34 mutant and Northern hybridization with probes which were specific with tRNA$^{Asp}$ or tRNA$^{Asn}$. The AspRS from A. ferrooxidans produced more Asp-tRNA than that from E. coli. Only aspS gene from A. ferrooxidans suppressed trpA34 strain in minimal media without tryptophan. Only AspRS from A. ferrooxidans showed mischarged Asp-tRNA$^{Asn}$ band. Therefore, AspRS from A. ferrooxidans is definitely ND-AspRS.

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Sonchus asper extract inhibits LPS-induced oxidative stress and pro-inflammatory cytokine production in RAW264.7 macrophages

  • Wang, Lan;Xu, Ming Lu;Liu, Jie;Wang, You;Hu, Jian He;Wang, Myeong-Hyeon
    • Nutrition Research and Practice
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    • v.9 no.6
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    • pp.579-585
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    • 2015
  • BACKGROUND/OBJECTIVES: Sonchus asper is used extensively as an herbal anti-inflammatory for treatment of bronchitis, asthma, wounds, burns, and cough; however, further investigation is needed in order to understand the underlying mechanism. To determine its mechanism of action, we examined the effects of an ethyl acetate fraction (EAF) of S. asper on nitric oxide (NO) production and prostaglandin-E2 levels in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. MATERIALS/METHODS: An in vitro culture of RAW264.7 macrophages was treated with LPS to induce inflammation. RESULTS: Treatment with EAF resulted in significant suppression of oxidative stress in RAW264.7 macrophages as demonstrated by increased endogenous superoxide dismutase (SOD) activity and intracellular glutathione levels, decreased generation of reactive oxygen species and lipid peroxidation, and restoration of the mitochondrial membrane potential. To confirm its anti-inflammatory effects, analysis of expression of inducible NO synthase, cyclooxygenase-2, tumor necrosis factor-${\alpha}$, and the anti-inflammatory cytokines IL-$1{\beta}$ and IL-6 was performed using semi-quantitative RT-PCR. EAF treatment resulted in significantly reduced dose-dependent expression of all of these factors, and enhanced expression of the antioxidants MnSOD and heme oxygenase-1. In addition, HPLC fingerprint results suggest that rutin, caffeic acid, and quercetin may be the active ingredients in EAF. CONCLUSIONS: Taken together, findings of this study imply that the anti-inflammatory effect of EAF on LPS-stimulated RAW264.7 cells is mediated by suppression of oxidative stress.

The Effect of Eungapbang-gagam on Thrombus Disease Related Factors and Oxidative Stress (은갑방가감(銀甲方加減)이 혈전병태유관인자(血栓病態有關因子)와 산화적손상(酸化的損傷)에 미치는 영향(影響))

  • Lee, Soo-Jeong;Kim, Song-Baeg
    • The Journal of Korean Obstetrics and Gynecology
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    • v.21 no.2
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    • pp.125-151
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    • 2008
  • Purpose: In this study, we investigated the anti-thrombotic efficacy of "Eungapbang-gagam(EGB)" currently used in clinical treatment of PID Methods: We studied inhibitory effect of platelet cohesion, suppressive effect of GPIIb/IIIa activity, inhibitory effect of $TXB_2$ and $PGE_2$ biosynthesis, and oxidative damage suppression effects of "EGB" in vitro. Also, suppression of pulmonary embolism and changes of related factors in dextran coagulation condition model were studied in vivo. Results: In this study, EGB extract showed dose-dependent inhibitory effect on platelet coagulation induced by ADP, epinephrine, collagen, arachidonic acid. Also it showed dose-dependent inhibition effect on GPIIb/IIIa activities compared to the control group. EGB extract significantly suppressed the decrease of speed of bloodstream caused by blood coagulation in dextran coagulation condition model and increased the number of platelets and amount of fibrinogen, and decreased the APTT in dextran coagulation condition model compared to the control group. EGB extract showed dose-dependent decrease of oxidative damages caused by DPPH and superoxide anion radicals, whereas dose-dependent increase of superoxide dismutase like activity was observed compared to the control group. Conclusion: We confirmed the anti-thrombosis and anti-oxidative efficacy of "Eungapbang-gagam". Various clinical applications of "Eungapbang-gagam" as well as use of data for the construction of EBM is anticipated.

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Ginsenoside compound K ameliorates palmitate-induced atrophy in C2C12 myotubes via promyogenic effects and AMPK/autophagy-mediated suppression of endoplasmic reticulum stress

  • Kim, Tae Jin;Pyun, Do Hyeon;Kim, Myeong Jun;Jeong, Ji Hoon;Abd El-Aty, A.M.;Jung, Tae Woo
    • Journal of Ginseng Research
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    • v.46 no.3
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    • pp.444-453
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    • 2022
  • Background: Compound K (CK) is among the protopanaxadiol (PPD)-type ginsenoside group, which produces multiple pharmacological effects. Herein, we examined the effects of CK on muscle atrophy under hyperlipidemic conditions along with its pro-myogenic effects. Further, the molecular pathways underlying the effects of CK on skeletal muscle have been justified. Methods: C2C12 myotubes were treated with palmitate and CK. C2C12 myoblasts were differentiated using CK for 4-5 days. For the in vivo experiments, CK was administered to mice fed on a high-fat diet for 8 weeks. The protein expression levels were analyzed using western blotting analysis. Target protein suppression was performed using small interfering (si) RNA transfection. Histological examination was performed using Jenner-Giemsa and H&E staining techniques. Results: CK treatment attenuated ER stress markers, such as eIF2a phosphorylation and CHOP expression and impaired myotube formation in palmitate-treated C2C12 myotubes and skeletal muscle of mice fed on HFD. CK treatment augmented AMPK along with autophagy markers in skeletal muscle cells in vitro and in vivo experiments. AMPK siRNA or 3-MA, an autophagy inhibitor, abrogated the impacts of CK in C2C12 myotubes. CK treatment augmented p38 and Akt phosphorylation, leading to an enhancement of C2C12 myogenesis. However, AMPK siRNA abolished the effects of CK in C2C12 myoblasts. Conclusion: These findings denote that CK prevents lipid-induced skeletal muscle apoptosis via AMPK/autophagy-mediated attenuation of ER stress and induction of myoblast differentiation. Therefore, we may suggest the use of CK as a potential therapeutic approach for treating muscle-wasting conditions associated with obesity.

Anti-tumor Effects of Soybeans and Fermented Soybean Paste

  • Lee, Sung-Lim;Kim, Jong-Gyu
    • Proceedings of the Korean Environmental Health Society Conference
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    • 2004.06a
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    • pp.182-184
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    • 2004
  • Oral cancer is the sixth most common cancer globally. The effects of several extracts from soybeans and Korean soybean paste (doen-jang) on the growth of human oral carcinoma cells in vitro were assessed. We prepared petroleum ether extract, ethyl acetate extract, chloroform extract, methanol extract, and water extract from soybeans and soybean paste. We used KB cell, which is an oral epidermoid carcinoma cell, and investigated proliferation of the tumor cells using MTT method. Each extract of soybean paste suppressed the KB cell proliferation. A dose-response relationship was observed between the level of ethyl acetate extract of soybean paste and its suppression of the cell proliferation. The effects of soybean extracts were lower than those of soybean paste extracts. The effects might be enhanced by the fermentation of soybeans. The results of this work indicate that extracts from soybeans and Korean soybean paste could have potential as anti-tumor substances.

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