• Title/Summary/Keyword: cell-based assay

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Potential Effect of Solvent Fractions of Taraxacum mongolicum H. on Protection of Gastric Mucosa (민들레(Taraxacum mongolicum H.) 추출분획물이 위장보호에 미치는 효능 평가)

  • Han, So-Heui;Hwang, Jung-Keun;Park, Soo-Nam;Lee, Kil-Hong;Ko, Kang-Il;Kim, Ki-Su;Kim, Ki-Ho
    • Korean Journal of Food Science and Technology
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    • v.37 no.1
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    • pp.84-89
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    • 2005
  • Mongolian dandelion (Taraxacum mongolicum H.) extracted with solvents having different polarities were prepared to investigate protection activity of gastric mucosa. Ethanol extracts were successively reextracted with methylene chloride, and n-butanol. Concentrations of total flavonoids and luteolin in n-butanol extracts were 27.75 and 1.14%, respectively, much higher than those of other solvent extracts. Results of anti-microbial acitivity test against Helicobacter pylori and urease inhibition test revealed n-butanol extract exerted higher inhibition (13.16%) than other solvent extracts. Based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, n-butanol extract showed the highest efficiency of free radical-scavenger activity among the extracts ($SC_{50}\;47{\mu}g/mL$). Erythrocytic photohemolysis tests, for the protection of cell membrane showed that half-life of human erythrocytes was increased by the addition of n-butanol extract $({\tau}_{50}=172min;C=5.0{\mu}g/mL)$. These results indicate n-butanol extract of Mongolian dandelion may be useful as a adjuvant for gastric diseases.

Characterization of Two Novel mAbs Recognizing Different Epitopes on CD43

  • Kim, Soseul;Hong, Jeong Won;Cho, Woon-Dong;Moon, Yoo Ri;Yoon, Sang Soon;Kim, Min-Young;Hong, Kwon Pyo;Lee, Yong-Moon;Yi, Jae Hyuk;Ham, Young Jun;Rah, Hyung Chul;Kim, Seung Ryul;Song, Hyung Geun
    • IMMUNE NETWORK
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    • v.14 no.3
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    • pp.164-170
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    • 2014
  • JL1, a specific epitope on CD43, is a potential biomarker for the diagnosis of acute leukemia. Although qualitative assays for detecting leukemia-specific CD43 exist, there is a need to develop quantitative assays for the same. Here, we developed two novel monoclonal antibodies (mAbs), 2C8 and 8E10, recognizing different epitopes on CD43. These clones are capable of pairing with YG5, another mAb against JL1 epitope, because they were selectively obtained using sandwich ELISA. Antigens recognized by 2C8 and 8E10 were confirmed as CD43 by western blotting using the CD43-hFC recombinant protein. When expression on various leukemic cell lines was investigated, 2C8 and 8E10 displayed a disparity in the distribution of the epitope. Enzyme assays revealed that these mAbs recognized a sialic acid-dependent epitope on CD43. Using normal thymus and lymph node paraffin-embedded tissues, we confirmed a difference in the epitopes recognized by the two mAbs that was predicted based on the maturity of the cells in the tissue. In summary, we developed and characterized two mAbs, 2C8 and 8E10, which can be used with YG5 in a sandwich ELISA for detecting leukemia-specific CD43.

Inhibitory Effects of High Concentrations of Estrogen, Progesterone and Tamoxifen on Proliferation of HeLa in Culture (배양된 HeLa 세포에서 고농도의 에스트로겐, 프로게스테론 및 타목시펜의 세포증식 억제효과)

  • Min, Gye-Sik
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1746-1751
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    • 2011
  • This study examined the effects of estrogen, progesterone and tamoxifen at different concentrations and treatment periods on proliferation of a human cervical carcinoma cell line, HeLa, in culture, based on MTT assay. Estrogen did not have an effect on the cellular proliferation in concentrations up to $1{\mu}g$/ml for treatment periods of between 2.5 and 6 days, but significantly inhibited proliferation at a higher concentration of $10{\mu}g$/ml in a progressive manner with increasing treatment periods. Also, treatment of HeLa with more than $10{\mu}g$/ml of progesterone for 2.5 days significantly inhibited proliferation and caused a concentration-dependent inhibition with 4 days of treatment. However, longer treatment with progesterone for 6 days abolished the concentration-dependent inhibitory effect on cellular proliferation observed with the 4-day treatment period. Furthermore, tamoxifen required a higher concentration ($100{\mu}g$/ml) than estrogen to bring about the inhibitory effect on the HeLa proliferation. These results suggest that high concentrations of estrogen, progesterone and tamoxifen may suppress proliferation of HeLa, and both the concentration and the treatment period may also influence their inhibitory effects on cellular proliferation.

A Study on Genetic Analysis and Extract Cytotoxicity of Scolopendra subspinipes multilans L. Koch (노랑머리왕지네의 유전학적(遺傳學的) 분석(分析) 및 약침액(藥鍼液)의 세포독성(細胞毒性)에 관한 연구(硏究))

  • Kim, Sung-Nam;Lim, Jeong-A;Lee, Sung-Yong;Hwang, Woo-Jun;Lee, Geon-Mok;Cho, Nam-Geun;Seo, Jung-Chul;Moon, Hyung-Cheol;Kim, Sung-Chul
    • Journal of Pharmacopuncture
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    • v.9 no.2
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    • pp.49-65
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    • 2006
  • Objective : The purpose of this study is to investigate nucleotide sequence and extract cytotoxicity of Scolopendrae corpus. The nature and taste of Scolopendrae corpus is hot, Warm and toxic, and the effect of this is dispelling wind, anti-spasmodic action and detoxication so it has been used for C.V.A, facial palsy, sensory disorder at extremities, wounds and arthritis. Methods : Scolopendrae corpus were collected by locality on the market. They were morphologically classified. Their nucleotide sequence was investigated and compared among them. In addition, the water-alcohol extract cytotoxicity of them was studied by MTT-based cytotoxicity assay. Results : It was shown that the each Scolopendrae corpus by locality is almost identical at genetic result and is identified as Scolopendra subspinipes mutilans L. Koch. Nucleotide sequence of Scolopendra subspinipes mutilans L. Koch in this study will help to discriminate other species of Scolopendrae corpus. The water-alcohol extract of Scolopendra subspinipes mutilans L. Koch did not induce cytotoxicity on Hep G2, L929 cell and peritoneal macrophages. Besides, it did not influence nitrite production of peritoneal macrophages. These results can be used as basic data for genetic discrimination with another species of scolopendrae corpus.

Inhibitory effect of Petalonia binghamiae on neuroinflammation in LPS-stimulated microglial cells (LPS에 의해 활성화된 미세아교세포에서 미역쇠 추출물의 신경염증 보호 효과)

  • Park, Jae Hyeon;Kim, Sung Hun;Lee, Sun Ryung
    • Journal of Nutrition and Health
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    • v.50 no.1
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    • pp.25-31
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    • 2017
  • Purpose: Neuroinflammation is mediated by activation of microglia implicated in the pathogenesis of neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease. Inhibition of neuroinflammation may be an effective solution to treat these brain disorders. Petalonia binghamiae is known as a traditional food, based on multiple biological activities such as anti-oxidant and anti-obesity. In present study, the anti-neuroinflammatory potential of Petalonia binghamiae was investigated in LPS-stimulated BV2 microglial cells. Methods: Cell viability was measured by MTT assay. Production of nitric oxide (NO) was examined using Griess reagent. Expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) was detected by Western blot analysis. Activation of nuclear factor ${\kappa}B$ ($NF-{\kappa}B$) signaling was examined by nuclear translocation of $NF-{\kappa}B$ p65 subunit and phosphorylation of $I{\kappa}B$. Results: Extract of Petalonia binghamiae significantly inhibited LPS-stimulated NO production and iNOS/COX-2 protein expression in a dose-dependent manner without cytotoxicity. Pretreatment with Petalonia binghamiae suppressed LPS-induced $NF-{\kappa}B$ p65 nuclear translocation and phosphorylation of $I{\kappa}B$. Co-treatment with Petalonia binghamiae and pyrrolidine duthiocarbamate (PDTC), an $NF-{\kappa}B$ inhibitor, reduced LPS-stimulated NO release compared to that in PB-treated or PDTC-treated cells. Conclusion: The present results indicate that extract of Petalonia binghamiae exerts anti-neuroinflammation activities, partly through inhibition of $NF-{\kappa}B$ signaling. These findings suggest that Petalonia binghamiae might have therapeutic potential in relation to neuroinflammation and neurodegenerative diseases.

Role of Rab11 on Membrane Trafficking of Rat Vanilloid Receptor, TRPV1 (바닐로이드 수용체 TRPV1의 막수송과정에서의 Rab11의 역할)

  • Um, Ki-Bum;Lee, Soon-Youl
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.12 no.7
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    • pp.3096-3102
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    • 2011
  • Vanilloid receptor, TRPV1 (transient receptor potential vanilloid 1) is a non-selective cation channel that responds to a variety of pain-eliciting material including capsaicin, pH, heat. Although, membrane trafficking of TRPV1 was not much known so far, TRPV1 was reported to interact with FIP3 (family of Rab11 interacting protein 3). FIP3 was identified as one of Rab11 interacting proteins that is recently reported important in membrane trafficking of several channel proteins directly or indirectly. Therefore, in this study, we examined the role of Rab11 in the membrane trafficking of TRPV1 using cell biological and biochemical techniques. Rab11 was found really colocalized with TRPV1 based on the result of confocal microscopy. However, GST-pulldown assay, one of biochemical technique, found that Rab11 did not interact with TRPV1. Although Rab11 does not interact with TRPV1 directly, we hypothesized that Rab11 is indeed involved in the membrane trafficking of TRPV1. In order to examine further the role of Rab11 in the membrane trafficking of TRPV1, the expression of TRPV1 on the membrane was examined when the expression of Rab11 was decreased down to about 50% by siRNA technique and found decreased significantly. From this result, we can conclude that Rab11 is involved in the membrane trafficking of TRPV1 in a way of including FIP3.

Downregulatory Effect of AGI-1120 $({\alpha}-Glucosidase Inhibitor)$ and Chaga Mushroom (Inonotus obliquus) on Cellular $NF-{\kappa}B$ Activation and Their Antioxidant Activity (AGI-1120과 차가버섯의 $NF-{\kappa}B$ 활성화 억제 및 항산화 효과)

  • Song, Hee-Sun;Lee, Young-Jong;Kim, Seung-Kyoon;Moon, Won-Kuk;Kim, Dong-Woo;Kim, Yeong-Shik;Moon, Ki-Young
    • Korean Journal of Pharmacognosy
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    • v.35 no.1 s.136
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    • pp.92-97
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    • 2004
  • Effect of AGI $({\alpha}-Glucosidase\;Inhibitor)-1120$, pine (Pinus densiflora) bark extract and Chaga mushroom (Inonotus obliquus) - and Chaga mushroom mycelium extracts on cellular $NF-{\kappa}B$ activation in malignant human keratinocytes (SCC-13) were evaluated to elucidate the possible correlation of $NF-{\kappa}B$ with antioxidant activity. The antioxidant activities of these natural products were examined in three different evaluation methods, i.e., lipid peroxidation value (POV) evaluation test, and 1,1diphenyl-2-picrylhydrazyl radical (DPPH) and nitric oxide (NO) scavenging test. In a cell-based $NF-{\kappa}B$ monitoring assay systern, all samples revealed the downregulatory profiles on the cellular $NF-{\kappa}B$ activity. AGI -1120 (1, 2 mg) and Chaga mushroom extract (0.05, 0.1 mg) downregulated the $NF-{\kappa}B$ activity in a dose-dependent manner. Chaga mushroom mycelium extract (5 mg) significantly inhibited the $NF-{\kappa}B$ activity (p<0.05). Although AGI-1120 and Chaga mushroom mycelium extract exhibited no antioxidant activities evaluated in pay, Chaga mushroom extract showed antioxidant in a dose-dependent manner at concentrations of $0.05{\sim}1$ mg. While AGI-1120 and Chaga mushroom extract possessed a relatively potential DPPH radical scavenging activity, the NO scavenging activity of Chaga mushroom extract $(SC_{50}:47\;{mu}g)$ was higher than the known antioxidant, vitamin C $(SC_{50}:77\;{mu}g)$. These results suggest that AGI-1120 and Chaga mushroom- and Chaga mushroom mycelium extracts may serve as an useful radical scavenging antioxidant agents with $NF-{\kappa}B$ inhibitory effect in human skin.

Remifentanil Protects Human Keratinocyte Through Autophagic Expression

  • Kim, Eok Nyun;Park, Chang Hoon;Woo, Mi Na;Yoon, Ji Young;Park, Bong Soo;Kim, Yong Ho;Kim, Cheul Hong
    • Journal of The Korean Dental Society of Anesthesiology
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    • v.14 no.2
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    • pp.101-106
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    • 2014
  • Background: Remifentanil, an ultra-short-acting mu-opioid receptor agonist, is unique from other opioids because of its esterase-based metabolism, minimal accumulation, and very rapid onset and offset of clinical action. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. However, the effects of remifentanil on human keratinocyte and autophagy have yet to be fully elucidated during hypoxia-reoxygenation. Here we investigated whether remifentanil confers protective effect against hypoxia-reoxygenation in human keratinocyte and, if so, whether autophagy mediates this effect. Methods: The human keratinocytes were cultured under 1% oxygen tension. The cells were gassed with 94% $N_2$, and 5% $CO_2$ and incubated for 24 h at $37^{\circ}C$. To determine whether the administration of affects human keratinocytes hypoxia-reoxygenation injury, cells were then exposed to various concentrations of remifentanil (0.01, 0.1, 0.5 and 1 ng/ml) for 2 h. After remifentanil treatment, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. Control group did not receive remifentanil treatment. Normoxia group did not receive hypoxia and remifentanil treatment for 36 h. 3-MA group was treated 3-methyladenine (3-MA) for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with MTT, showing the mitochondrial activity of living cells. Cells were stained with fluorescence and analyzed with Western blot analysis to find out any relations with activation of autophagy. Results: Prominent accumulation of autophagic specific staining MDC was observed around the nuclei in RPT group HaCaT cells. Similarly, AO staining, red fluorescent spots appeared in RPT group HaCaT cells, while the Normoxia, control and 3-MA groups showed mainly green cytoplasmic fluorescence. We here examined activation of autophagy related protein under H/R-induced cells by Western blotting analysis. Atg5, Beclin-1, LC3-II (microtubule-associated protein 1 light chain 3 form II) and p62 was elevated in RPT group cells. But they were decreased when autophagy was suppressed by 3-MA (Fig. 5). Conclusions: Although the findings of this study are limited to an in vitro interpretation, we suggest that remifentanil may have a beneficial effect in the recovery of wound from hypoxia-reoxygenation injury.

Effect of Dendropanax morbifera extract Addition on Chungkukjang on Immune Response of Mice (황칠 추출물을 첨가한 청국장이 마우스의 면역기능에 미치는 효과)

  • Kim, Jin-Sol;Cheong, Min-Ju;Chung, Kyoung-A;Song, Seon-Young;Lee, Hyun-Hwa
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.6
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    • pp.430-438
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    • 2019
  • This study is to investigate increase immunomodulatory activity of Chungkukjang added with Dendropanax morbifera extract. There are four groups divided that Control, Dendropanax morbifera extract group, Chungkookjang group and Chungkukjang added to Dendropanax morbifera extract group which of five mouses each group. Their immunological characteristics were compared with either general Chungkukjang or non treated group in the tested male ICR mice. There were no significant difference in body weight and organ weight. In the plaque forming cell assay, Dendropanax morbifera extract 250 and 500 mg/kg groups measured significant increase over the control group. The values of lymphocytes were significantly increased in the Dendropanax morbifera extract 500 mg/kg group campared with the control group. In the general Chungkukjang 400 mg/kg added with Dendropanax morbifera extract 500 mg/kg group, total serum immunoglobulin G concentration was significantly higher than the control group and their spleen tissues observed proliferation of white puls. These results demonstrated that Chungkukjang Added with Dendropanax morbifera extract was provided enhance of immunomodulatory activity and suggest that it can be used as various functional foods, based on foods promote immune system health.

A Study on the Antioxidant and MMPs Protein Expression Inhibitive Effect of Punica granatum L. Extract and Its Stabilization with Liquid Crystal Emulsion (석류추출물의 항산화와 MMPs 단백질 발현 억제 및 액정 유화물에서의 안정화에 관한 연구)

  • Roh, Jin-Sun;Yeom, Hyeon-Ji;Oh, Min-Jeong;Lee, Jin-Young
    • Journal of Life Science
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    • v.31 no.2
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    • pp.164-174
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    • 2021
  • This study confirmed the potential of Punica granatum L. extract for functional activity verification and cosmetic development. The electron-donating ability of Punica granatum L. extract was shown 60.6% at a 1,000 ㎍/ml concentration. Its ABTS+ radical scavenging ability was shown 93.9% at a 1,000 ㎍/ml concentration. Additionally, the inhibitive effects of elastase and collagenase inhibition effects were measured as 30% and 47.2%, respectively, at a 1,000 ㎍/ml concentration. To determine the effect of Punica granatum L. extract on the proliferation of fibroblasts (CCD-986sk), cell viability was measured using a 3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide (MTT) assay. As a result, survival rates of 130% or higher at a 500 ㎍/ml concentration or less were confirmed. According to the results of Western blot with Punica granatum L. extract, the expression inhibition rates of matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-2 (MMP-2), and matrix metalloproteinase-3 (MMP-3) were decreased by 23.2%, 81.9%, and 69.2%, respectively, at a 100 ㎍/ml concentration. Based on the results above, O/W liquid crystal cream with 0.1% Punica granatum L. extract was prepared. The stabilities were tested at 4, 25, 45, and 50℃. By checking the pH, change over time, and stability by temperature, it was confirmed that all were stable for one month. Thus, Punica granatum L. extract shows potential as a natural material for cosmetics.