• Title/Summary/Keyword: Expression and secretion

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Comparison of Marker Components and Biological Activities of Socheongryong-tang by Different Extract Methods (소청룡탕 제조방법에 따른 성분 및 생리활성 비교)

  • Baek, Ka Yeon;Kim, Se Jin;Kim, Jong Beom;Lee, Jeong Hyeon;Moon, Sung Ok;Lee, Hwa Dong
    • Korean Journal of Pharmacognosy
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    • v.49 no.4
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    • pp.349-361
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    • 2018
  • Socheongryong-tang (SCRT) was one of the major traditional herbal medicines wildly used in the treatment of respiratory disease. SCRT is being commercially produced in the form of mix extracts powder and soft dry extract by different extract methods in the Korean Herbal Pharmacopeia (KHP). In this study, the contents of marker components and biological activities of the SCRT mix extract powder were compared with those of the SCRT decoction. To analyze the marker components of SCRT, nine marker from eight herbal preparations were chosen. And the method using high performance liquid chromatography (HPLC) with diode-array detector method was established for the simultaneous analysis. Method validation was accomplished by linearity, precision test, and recovery test. The contents of nine marker components in this extract was ascertained by ratio. The biological activities were examined the effect of SCRT on anti-oxidation and pro-inflammation mediated by LPS-stimulation. We confirmed that both of SCRT mix extrct powder and decoction have the similar contents on total polyphenol and flavonoid and inhibited the secretion of nitric oxide (NO), $IL-1{\beta}$, IL-6, tumor necrosis factor $(TNF)-{\alpha}$ and the expression of iNOS, COX-2, $IL-1{\beta}$, IL-6, $TNF-{\alpha}$. These results suggest that SCRT mix extract powder and decoction have a significant correlation.

Evaluation on Pharmacological Effects and Component Analysis of Hwanggeumjakyak-tang Formulation for Tablet (황금작약탕(黃芩芍藥湯)을 함유하는 정제 개발과 성분분석 및 약리효능 평가)

  • Kim, Se-Jin;Lee, Ji-Beom;Choi, Hye-Min;Lee, Ha-Yeong;Kim, Min-Ju;Lee, Mi-Hwa;Kim, Jung-Ok;Lee, Hwa-Dong
    • Korean Journal of Pharmacognosy
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    • v.49 no.4
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    • pp.362-371
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    • 2018
  • Hwanggeumjackyak-tang (HJT) composed of Scatellaria baicalensis Georgi, Paeonia Lactiflora and Glycyrrhizae uralensis Fischer is a traditional Korean herbal medicine widely used for acute enteritis. In order to develop the tablet formulation of HJT, evaluation of the flow properties, thickness, diameter, hardness, friability and disintegration was carried out on four HJT granules according to mixed content of seven additives. Simultaneous analysis used HPLC method was performed of HJT tablet and was determined of the seven marker components; Albiflorin, Paeoniflorin, Liquiritin, Baicalin, Baicalein, Glycyrrhizic acid and Wogonin. The biological activities were examined the effect of HJT on anti-oxidation and pro-inflammation mediated by LPS-stimulation. We confirmed that both of HJT-Decoction (HJT-D) and HJT-Formulation (HJT-F) have the similar contents on total polyphenol and flavonoid and inhibited the secretion of nitro oxide (NO), interleukin $(IL)-1{\beta}$,interleukin (IL)-6, tumor necrosis factor $(TNF)-{\alpha}$ and the expression of nitric oxide synthase (iNOS), $IL-1{\beta}$, IL-6, $TNF-{\alpha}$. Therefore, the developed formulation for tablet of HJT would provide chemically and biologically the same effect compared with decoction of HJT.

Effect of long-chain inorganic polyphosphate treated with wheat phytase on interleukin 8 signaling in HT-29 cells

  • An, Jeongmin;Cho, Jaiesoon
    • Animal Bioscience
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    • v.35 no.6
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    • pp.892-901
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    • 2022
  • Objective: This study was performed to investigate the potential effect of wheat phytase on long-chain inorganic polyphosphate (polyP)-mediated interleukin 8 (IL-8) signaling in an intestinal epithelial cell line, HT-29 cells. Methods: Cell viability and the release of the pro-inflammatory cytokine IL-8 in HT-29 cells exposed to polyP1150 (average of 1,150 phosphate residues) treated with or without wheat phytase were measured by the EZ-CYTOX kit and the IL-8 ELISA kit, respectively. Also, the activation of cellular inflammatory factors NF-κB and MAPK (p38 and ERK 1/2) in HT-29 cells was investigated using ELISA kits. Results: PolyP1150 negatively affected the viability of HT-29 cells in a dose-dependent manner. However, 100 mM polyP1150 dephosphorylated by wheat phytase increased cell viability by 1.4-fold over that of the intact substrate. Moreover, the 24 h exposure of cells to enzyme-treated 50 mM polyP1150 reduced the secretion of IL-8 and the activation of NF-κB by 9% and 19%, respectively, compared to the intact substrate. PolyP1150 (25 and 50 mM) dephosphorylated by the enzyme induced the activation of p38 MAPK via phosphorylation to 2.3 and 1.4-fold, respectively, compared to intact substrate, even though it had little effect on the expression of ERK 1/2 via phosphorylation. Conclusion: Wheat phytase could attenuate polyP1150-induced IL-8 release in HT-29 cells through NF-κB, independent of MAP kinases p38 and ERK. Thus, wheat phytase may alleviate inflammatory responses including hypercytokinemia caused by bacterial polyP infection in animals. Therefore, wheat phytase has the potential as an anti-inflammatory therapeutic supplement in animal husbandry.

Change of Protein Patterns in Uterine Fluid during Estrus Cycle in Pigs (돼지에서 발정 주기 동안 자궁액 내의 단백질 패턴의 변화)

  • Lee, Yeon-Ju;Song, Eun-Ji;Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.103-108
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    • 2013
  • An uterus is female reproductive tract organ that affected estrus cycle. During a various changes occur at uterus in estrus cycle, one of them is body fluids secretion be called uterine fluid. Therefore, the objective of this study was to investigate the changes of protein patterns using two-dimensional gel electrophoresis in uterus fluids during the follicular and luteal phases in estrus cycle of pigs. In changes of protein spots were confirmed during the follicular and luteal phases. The 136 spots were expressed in follicular phase, the 57 spots of them showed reproducibility. On the other hand, the 140 spots were expressed in luteal phase, the 73 spots of them showed reproducibility. Also, spots expressed in follicular phase were number 69 and 94 spots and spots expressed in luteal phase only were number 156, 157, 184~187, 190 and 191 spots. The spots which of higher expression levels in the luteal phase than in follicular phase were number 76 and 79 spots. In conclusion, the spots expressed in follicular and luteal phases were confirmed with difference levels and these differences are function of RNA resolving, protein synthesis and cytoskeletal architecture.

Inhibition of melanogenesis by sodium 2-mercaptoethanesulfonate

  • Kim, Jeong-Hwan;Oh, Chang-Taek;Kwon, Tae-Rin;Kim, Jong Hwan;Bak, Dong-Ho;Kim, Hyuk;Park, Won-Seok;Kim, Beom Joon
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.2
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    • pp.149-156
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    • 2020
  • Sodium 2-mercaptoethanesulfonate (mesna) is a protective agent that is widely used in medicine because of its antioxidant effects. Recently, reactive oxygen species (ROS) were shown to increase pigmentation. Thus, ROS scavengers and inhibitors of ROS production may suppress melanogenesis. Forkhead box-O3a (FoxO3a) is an antimelanogenic factor that mediates ROS-induced skin pigmentation. In this study, we aimed to investigate the whitening effect of mesna and the signaling mechanism mediating this effect. Human melanoma (MNT-1) cells were used in this study. mRNA and protein expression were measured by real-time quantitative PCR and Western blotting analysis to track changes in FoxO3a-related signals induced by mesna. An immunofluorescence assay was performed to determine the nuclear translocation of FoxO3a. When MNT-1 melanoma cells were treated with mesna, melanin production and secretion decreased. These effects were accompanied by increases in FoxO3a activation and nuclear translocation, resulting in downregulation of four master genes of melanogenesis: MITF, TYR, TRP1, and TRP2. We found that mesna, an antioxidant and radical scavenger, suppresses melanin production and may therefore be a useful agent for the clinical treatment of hyperpigmentation disorders.

Liripois Tuber contributes to the chemotaxis of eosinophils and secretion of cytokines in A549 human epithelial cells (맥문동(麥門冬)이 천식유발 cytokine 분비와 호산구 chemotaxis에 미치는 영향)

  • Jung, Hae-Jun;Jung, Gee-Jae;Jung, Sung-Ki;Rhee, Hyung-Koo
    • The Journal of Korean Oriental Chronic Disease
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    • v.10 no.1
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    • pp.1-20
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    • 2005
  • Background and Objective: The importance of the presence of eosinophils in the airways of patients with fetal asthma has long been recognized, but the mechanism by which these cells are recruited and retained in the lung are only now being elucidated. Eotaxin is a potent and specific eosinophil chemoattractant that is mobilized in the respiratory epithelium after allergic stimulation. Material and Methods: We used water extracts of Liripois Tuber and pulmonary epithelial cell lines A549(human typeII-like epithelial cells) and human eosinophils. We estimated cytotoxic effects of Liripois Tuber via MTS assay, and estimated the effects of Liripois Tuber on chemokines from prestimulated A549 cells by sandwich ELISA and RT-PCR. We conducted chemotaxis assay on prestimulated eosinophils treated with Liripois Tuber. Result: In this study we demonstrated that $TNF-{\alpha}$, IL-4 and $IL-1{\beta}$ induced the accumulation of chemokines mRNA in the pulmonary epithelial cell lines A549 in dose-dependent manner. Chemokines were inhibited by Liripois Tuber in dose-dependent manner. The eosinophil migration was inhibited at high concentration of Liripois Tuber. Conculusion: These findings indicate that the supression of the expression of chemokines can be accomplished by Liripois Tuber treatment, raising the possibility that Liripois Tuber might be of therapeutic value in diseases such as asthma.

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Decreased Expression of Phospholipase C-$\beta$2 in Human Platelets with Impaired Function

  • Lee, Sang-Bong;A. Koneti Rao;Lee, Kweon-Haeng;Xu Yang;Bae, Yun-Soo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.11a
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    • pp.75-84
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    • 1996
  • Platelets from a patient with a mild inherited bleeding disorder and abnormal platelet aggregation and secretion show reduced generation of inositol 1,4,5-trisphosphate (IP$_3$), mobilization of intracellular Ca$\^$2+/, and phosphorylation of pleckstrin in response to several G protein mediated agonists, suggesting a possible defect at the level of phospholipase C (PLC) activation. A procedure was developed that allows quantitation of platelet PLC isozymes. After fractionation of platelet extracts by high-performance liquid chromatography, seven, out often known PLC isoforms were detected by immunoblot analysis. The amount of these isoforms in normal platelets decreased in the order PLC-${\gamma}$2 > PLC-${\beta}$2 > PLC-${\beta}$3 > PLC-${\beta}$l > PLC-${\gamma}$ > PLC-$\delta$1 > PLC-${\beta}$4. Compared with normal platelets, platelets from the patient contained approximately one-third the amount of PLC-${\beta}$2, whereas PLC-${\beta}$4 was increased threefold. These results suggest that the impaired platelet function in the patient in response to multiple G protein mediated agonists is attributable to a deficiency of PLC-${\beta}$2. They document for the first time a specific PLC isozyme deficiency in human platelets and provide an unique opportunity to understand the role of different PLC isozymes in normal platelet function.

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Zoledronate(Zometa(R))inhibits the formation of osteoblast in rat osteoblastic cell line UMR-106 (Zoledronate이 UMR-106 세포의 증식과 조골세포 형성에 미치는 영향)

  • Jeong, Ki-Hoon;Ryu, Dong-Mok;Jee, Yu-Jin;Lee, Deok-Won;Lee, Hyun-Woo
    • The Journal of the Korean dental association
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    • v.46 no.10
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    • pp.623-632
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    • 2008
  • Purpose : The purpose of this study is to identify the effect of zoledronate(Zometa(R)), which is most common nitrogen containing bisphosphonate, on survival, proliferation, and differentiation of osteoblast. Material & Methods: Twenty four cell culture plates containing essential medium were seeded with UMR-106 cell lines, at density of 5 x $10^4 cells per plates. Each plates were incubated with 5% $CO^2 incubator at $37^{\circ}C$. Starting from 2 days after incubation, cell culture medias were replaced, and added with osteogenesis induction media and 0, 0.01, 0.1, 0.5, 1, $3\muM$ of zoledronate(Zometa(R)), every 2 days, for 12 days. Control group was plates not added with zoledronate($0\muM$), and experiment group were plates added with different concentration of zoledronates(0, 0.01, 0.1, 0.5, 1, $3\muM$). Mature osteoblasts were identified with Alizarine Red staining, and protein samples were collected. Optical density was determined at wavelength of 405nm with ELISA reader. For viability analysis, cells were harvested and incubated with propidium iodide, and analysed with flow cytometry. Western blot technique was used to analyse Runx2 protein of osteoblast. Results : Secretion of bone matrix decreased as zoledronate concentration increased, and zoledronate did not effect survival rate of UMR-106 cells when measured with flow cytometer. Expression of Runx2 protein was inhibited as zoledronate concentration increased. Conclusion : From the results, we were able to identify that increase of zoledronate concentration inhibited differentiation of UMR-106 cell to osteoblast, without effecting quantity or survival rate.

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The Effects of Verapamil on Growth and Apoptosis of Keloid Fibroblast (Verapamil이 켈로이드 섬유모세포의 성장과 세포자멸사에 미치는 영향)

  • Park, Jung-Min;Lee, Keun-cheol;Kim, Seok-Kwun;Bae, Hae-Rahn;Rha, Seo-Hee
    • Archives of Plastic Surgery
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    • v.32 no.5
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    • pp.625-635
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    • 2005
  • In this study, the effects of verapamil on growth rate, apoptosis, production of transforming growth factor (TGF-${\beta}$) and fibronectin were evaluated in keloid and normal human dermal fibroblasts. Both fibroblasts were primarily cultured from earlobe keloids of three female patients and treated with various concentrations of verapamil. Cell toxicity was assessed by MTT assay, growth rate and apoptosis by FACS, and the production of TGF-${\beta}$ and fibronectin by ELISA and Western blot, respectively. In the $MTT_{50}$, the cell growth was more suppressed in keloid fibroblasts. In the $MTT_{90}$, cell growth was more stimulated in normal fibroblasts. No significant effect appeared on TGF-${\beta}$ expression but an increase in extracellular fibronectin secretion was found in keloid fibroblasts. Keloid fibroblasts responded to verapamil more sensitively, and the percentage of apoptosis was higher at the $MTT_{50}$l. In brief, verapamil had growth-inhibitory effect with inducing apoptosis at the $MTT_{50}$, but rather growth-stimulatory effect at the $MTT_{90}$. The biphasic effect of verapamil depending on the dose might explain one of the reasons of relapse after keloid treatment with verapamil. Clinical application with high concentration (2.5 mg/ml) is advised unless excessive dosage is used.

Anti-inflammatory effects of Hataedock with Coptidis Rhizoma and Glycyrrhiza Uralensis on Allergic Rhinitis through Regulating IL-4 Activation (알레르기성 비염에서 황련-감초 하태독법의 IL-4활성 조절을 통한 항염증효과)

  • Jung, A Ram
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.33 no.2
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    • pp.116-122
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    • 2019
  • The aim of this study is to evaluate the anti-inflammatory effect of Hataedock treatment using Coptidis Rhizome and Glycyrrhiza Uralensis (CG) mixed extract in allergic rhinitis induced NC/Nga mice. We divided NC/Nga mice into 3 groups as follows; allergic rhinitis-induced group after CG Hataedock treatment (CGT, n=10), no treatment group (Ctrl), allergic rhinitis elicited group (ARE). To induce allergic rhinitis, NC/Nga mice of 3 weeks age were sensitized on 7, 8 and 9week by Ovalbumin (OVA) antigen in intranasal space. Hataedock using CG extract was administered on week 3 in allergic rhinitis-induced group (CGT) after Hataedock treatment. To identify distribution of Interlukin (IL)-4, Cluster of differentiation 40 (CD40), high-affinity IgE receptor ($Fc{\varepsilon}RI$), substance P, Matrix metallopeptidase 9 (MMP-9), Nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) p65, Inducible nitric oxide synthase (iNOS) and Cycloxygenase-2 (COX-2), we used histological examination. CGT significantly inhibited IL-4 and CD40 response compared with ARE. The reduction of Th2 cytokine expression decreased inflammatory mediators such as $Fc{\varepsilon}RI$, substance P, MMP-9, $NF-{\kappa}B$ p65, iNOS and COX-2. Such immunological improvement induced reduction of respiratory epithelial damage and mucin secretion in goblet cell. These results indicate that Hataedock treatment suppresses allergic rhinitis through modulating of Th2 responses and diminishing various inflammatory mediators in nasal mucosal tissue. It might have potential applications for prevention and treatment of allergic rhinitis.