• Title/Summary/Keyword: mRNA 발현

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Inhibitory Effect on Kaempferia Parviflora Ethanol Extract of IL-1β Induced Inflammation and MMP Expression in CHON-001 Cells (흑생강 추출물의 CHON-001 세포에서의 IL-1β로 유도된 염증과 MMPs 발현)

  • Jeong Ah Lee;Hye Min Seol;Seong Un Jeong;Jae Hyeon Yoon;Jeong Soo Bae;Tae Hee Kim;Hyeong Soo Kim
    • Journal of Life Science
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    • v.34 no.8
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    • pp.558-566
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    • 2024
  • The potential therapeutic effects of Kaempferia Parviflora ethanol extract (KPE) on osteoarthritis were investigated using the human chondrocyte cell line (CHON-001) to explore its application in functional foods. The CHON-001 cells were pre-treated with either 1 ㎍/ml or 5 ㎍/ml of KPE before exposure to 10 ng/ml of IL-1β to induce osteoarthritis. Results showed that KPE treatment significantly suppressed IL-1β-induced TNF-α production by 66% and 50% at concentrations of 1 ㎍/ml and 5 ㎍/ml KPE, respectively. In addition, COX-2 protein expression was reduced by 26% and 34% compared to control levels. The preservation of chondrocyte-specific matrix proteins, aggrecan, and collagen type II, was notable, with aggrecan and mRNA levels maintained by 5% and 8%, and collagen II levels preserved by 62% and 47% at the same KPE concentrations. This preservation is likely due to the reduced expression of MMP1 and MMP13, enzymes responsible for matrix protein degradation. Overall, the current results suggest that KPE may protect chondrocytes from IL-1β-induced osteoarthritis by suppressing TNF-α production and COX-2 expression while preserving critical matrix proteins like aggrecan and collagen II by suppressing the expressions of their degrading enzymes (MMP-1 and MMP-13). Therefore, KPE holds promise as a candidate for developing functional foods aimed at reducing osteoarthritis.

Effects of Coenzyme Q10 on the Expression of Genes involved in Lipid Metabolism in Laying Hens (Coenzyme Q10 첨가 급여가 산란계의 지방대사 연관 유전자 발현에 미치는 영향)

  • Jang, In Surk;Moon, Yang Soo
    • Korean Journal of Poultry Science
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    • v.43 no.1
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    • pp.47-54
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    • 2016
  • The aim of this study was to investigate the expression patterns of key genes involved in lipid metabolism in response to dietary Coenzyme Q10 (CoQ10) in hens. A total of 36 forty week-old Lohmann Brown were randomly allocated into 3 groups consisting of 4 replicates of 3 birds. Laying hens were subjected to one of following treatments: Control (BD, basal diet), T1 (BD+ CoQ10 100 mg/kg diet) and T2 (BD+ micellar of CoQ10 100 mg/kg diet). Birds were fed ad libitum a basal diet or the basal diet supplemented with CoQ10 for 5 weeks. Total RNA was extracted from the liver for quantitative RT-PCR. The mRNA levels of HMG-CoA reductase(HMGCR) and sterol regulatory element-binding proteins(SREBP)2 were decreased more than 30~50% in the liver of birds fed a basal diet supplemented with CoQ10 (p<0.05). These findings suggest that dietary CoQ10 can reduce cholesterol levels by the suppression of the hepatic HMGCR and SREBP2 genes. The gene expressions of liver X receptor (LXR) and SREBP1 were down regulated due to the addition of CoQ10 to the feed (p<0.05). The homeostasis of cholesterol can be regulated by LXR and SREBP1 in cholesterol-low-conditions. The supplement of CoQ10 caused a decreased expression of lipid metabolism-related genes including $PPAR{\gamma}$, XBP1, FASN, and GLUTs in the liver of birds (p<0.05). These data suggest that CoQ10 might be used as a dietary supplement to reduce cholesterol levels and to regulate lipid homeostasis in laying hens.

Induction of c-Jun Expression by Breast Cancer Anti-estrogen Resistance-3 (BCAR3) in Human Breast MCF-12A Cells (정상적인 인간유방상피세포인 MCF-12세포에서 유방암 항에스토젠 내성인자-3 (BCAR3)에 의한 c-Jun 발현 유도 연구)

  • Oh, Myung-Ju;Kim, Ji-Hyun;Jhun, Byung Hak
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1383-1391
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    • 2016
  • Anti-estrogen drugs such as tamoxifen have been used for treating patients with ER-positive, early breast cancer. However, resistance to anti-estrogen treatment is inevitable in most patients. Breast cancer anti-estrogen resistance-3 (BCAR3) has been identified as the protein responsible for the induction of tamoxifen resistance in estrogen-dependent human breast cancer. We have previously reported that BCAR3 regulates the cell cycle progression and the signaling pathway of EGF and insulin leading to DNA synthesis. In this study, we investigated the functional role of BCAR3 in regulating c-Jun transcription in non-tumorigenic human breast epithelial MCF-12A cells. A transient transfection of BCAR3 increased both the mRNA and protein of c-Jun expression, and stable expression of BCAR3 increased c-Jun protein expression. The overexpression of BCAR3 directly activated the promoter of c-jun, AP-1, and SRE but not that of $NF-{\kappa}B$. Furthermore, single-cell microinjection of BCAR3 expression plasmid in the cell cycle-arrested MCF-12A cells induced c-Jun protein expression, and co-injection of dominant negative mutants of Ras, Rac, and Rho suppressed the transcriptional activity of c-Jun in the presence of BCAR3. Furthermore, stable expression of BCAR3 increased the proliferation of MCF-12A cells. The microinjection of inhibitory materials such as anti-BCAR3 antibody and siRNA BCAR3 inhibited EGF-induced c-Jun expression but did not affect IGF-1 induced upregulation of c-Jun. Taken together, we propose that BCAR3 plays a crucial role in c-Jun protein expression and cell proliferation and that small GTPases (e.g., Ras, Rac, and Rho) are required for the BCAR3-mediated activation of c-Jun expression.

Inhibitory Effects of Chios Mastic Gum on Gastric Acid Secretion by Histamine-Related Pathway in a Rat Model and Primary Parietal Cells (위염 동물모델과 위 벽세포에서 히스타민 경로를 통한 매스틱검(Chios Mastic Gum)의 위산 분비 억제효과 및 기전 연구)

  • Nam, Da-Eun;Kim, Ok Kyung;Shim, Tae Jin;Lee, Jum Kyun;Hwang, Kwon-Tack
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.10
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    • pp.1500-1509
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    • 2014
  • The object of this study was to investigate the inhibitory effects of chios mastic gum (MG) on gastric acid secretion in an ethanol-induced SD rat model and primary parietal cells. Rats were randomly divided into four groups: Vehicle (normal group), Control (treated with ethanol), MG50 (treated with ethanol and mastic gum at 50 mg/kg b.w), MG100 (treated with ethanol and mastic gum at 100 mg/kg b.w). Groups treated with both MG50 and MG100 showed attenuation of gastric mucosal injury, sub-epithelial loss, hemorrhaging, and gastric juice secretion. We also examined the acidity of gastric juice during gastric injury. Oral administration of both MG50 and MG100 significantly decreased acidity of gastric juice by % and %, respectively. To examine the stimulatory factors related to gastric acid secretion, mRNA expression levels of H2r, M3r, CCK2r, and $H^+/K^+$ ATPase were measured by real-time PCR. Compared with a vehicle group, mRNA expression levels of H2r, CCK2r, and $H^+/K^+$ ATPase clearly increased in the control group. However, levels of H2r, CCK2r, and $H^+/K^+$ ATPase slightly but significantly decreased in MG-treated groups compared with control. Blood level of histamine significantly decreased in MG-treated groups, which indicates the involvement of MG on in histamine-related acid secretion. To identify the mode of action of MG in regulating histamine-related pathways, intracellular level of cAMP and mRNA levels of H2r, M3r, CCK2r, and $H^+/K^+$ ATPase were measured in primary parietal cells. While mRNA levels of M3r and CCK2r remained unchanged, levels of H2r and $H^+/K^+$ ATPase significantly decreased upon MG treatment. Subsequently, intracellular levels of cAMP decreased. These results suggest that mastic gum has the ability to inhibit gastric acid secretion by regulating a histamine-related pathway.

Effects of the Draronis sanguis on Antioxidation and MMP-1 Expression in Human Dermal Fibroblast (혈갈(Draconis Sanguis)의 항산화와 사람섬유아세포에서 MMP-1 발현저해 효과)

  • Sim Gwan Sub;Kim Jin Hui;Kim Jin Hwa;Lee Dong Hawn;Park Sung Min;Lee Bum Chun;Pyo Hyeong Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.439-444
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    • 2004
  • UV irradiation produces free radicals and related reactive oxygen species (ROS), and these are injury to all most of organisms of skin cells and extracellular matrix (ECM). In addition, free radicals and ROS stimulate the overexpression of matrix metalloproteinases (MMPs) that can degrade most components of ECM such as collagen. Since collagen constitutes almost of skin connective tissue, their disarrangement causes wrinkle formation and droop of skin. Therefore, scavenging activity on free radicals, ROS and suppression of MMP-1 is expected to prevent skin photoaging. In this study, to investigate the relationship between photoaging and Draconis sanguis, we examined the effects of antioxidant, in vitro MMP inhibition and expression of UVA-induced MMP-1 in human dermal fibroblasts. Draconis sanguis was found to show scavenging activities of radicals and ROS with the $IC_{50}$ values of $183{\;}{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $30{\;}{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. Draconis sunguis inhibited the activities of MMP-1 in a does-dependent manner and the $IC_{50}$ value calculated from semi-log plots was $200{\;}{\mu}g/mL$. Also, UVA induced MMP expression was reduced $74\%$ by treatment with Draconis sanguis, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Therefore Draconis sanguis was able to significantly inhibit MMP expression in protein and mRNA level. All these results suggested that Draconis sanguis may act as an anti-photoaging agent by antioxidation and reducing UVA-induced MMP-1 production.

Genetic Transformation of Lettuce (Lactuca sativa L.) with Agrobacterium tumefaciens (Agrobacterium tumefaciens에 의한 상추 (Lactuca sativa L.)의 형질전환)

  • 최언옥;양문식;김미선;은종선;김경식
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.55-58
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    • 1994
  • Agrobacterium tumefaciens LABA4404 harboring plant binary vector, pBI121, was used for genetic transformation of lettuce (Lactuca sativa t.). Cotyledon segments were infected with A. tumefaciens LBA4404 by cocultivation method and regenerated. Regenerated letture was subject to molecular analyses for integration into plant nuclear genome and expression of ${\beta}$-glucumnidase (GUS) gene. Southern and Northern blot analyses demonstrated that GUS gene was integrated into plant nuclear genome and expressed into its mRNA. The expression of GUS gene into its protein was confirmed by specetrophotometric assay of GUS activity.

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Up-regulation of dynamin-2 gene expression in Ras-transformed cells (Ras에 의해 암화된 세포에서 dynamin-2의 발현 촉진)

  • Yoo, Ji-Yun
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.375-380
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    • 2007
  • Dynamin plays a key role in the scission event common to various types of endocytosis. It has been previously reported that the SH3 domain-mediated association of Grb2 with dynamin-2 was dominantly found in Ras transformed cells. However, whether this association results from the increased expression of dynamin-2 and Grb2 in Ras transformed cells or not is still unknown. So in this study we first analyzed the expression levels of dynamin-2 and Grb2 and found that the expression of dynamin-2 protein was dramatically increased in Ras-transformed NIH3T3 (NIH3T3(Ras)) cells. Furthermore competitive PCR data revealed that the mRNA transcripts for dynamin-2 were increased about 100-fold in NIH3T3(Ras) compared to those of NIH3T3 cells. However, the protein level and mRNA transcript of Grb2 were not changed in these two cells. We also examined promoter activity of dynamin-2 in NIH3T3(Ras) cells and suggest the existence of Ras-responsive sequence in promoter region -300 to -200.

The Effect of Dexamethasone on Gene Expression and Total Amount of Surfactant Protein A (스테로이드제가 Surfactant Protein A의 유전자 발현과 총단백량에 미치는 영향에 관한 실험적 연구)

  • Lim, Byung Sung;Sohn, Jang Won;Yang, Seok Chul;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.4
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    • pp.395-404
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    • 2002
  • Background : Surfactant protein A (SP-A) is important for regulating surfactant secretion, synthesis and recycling. However, It's regulation in vivo is unclear. SP-A has important roles in regulating surfactant metabolism as well as determining its physical properties. Glucocorticoid accelerates the morphologic differentiation of epithelial cells into type II cells and increase the rate of phosphatidylcholine synthesis. Methods : The authors investigated the effects of glucocorticoid on the accumulation of mRNA encoding SP-A and SP-A protein content. Adult rats were given various doses of subcutaneous dexamethasone and sacrificed after 24 hours and one week. SP-A mRNA was measured using a filter hybridization method. The lung SP-A protein content was determined using a double sandwich ELISA assay with polyclonal antiserum raised in rabbits against purified rat SP-A. Results : 1) The accumulation of SP-A mRNA in the dexamethasone treated group 24 hours after 0.2 mg/kg dexamethasone treatment was increased 38.8% compared to the control group. 2) The accumulation of SP-A mRNA in the dexamethasone treated group 1 week after 2 mg/kg dexamethasone treatment was 49.7% higher than the control group(P<0.01). 3) The total lung SP-A level was not altered after 24 hours by the 0.2mg/kg treatment. The total lung SP-A content one week after 2mg/kg dexamethasone administration was 373.7% higher than the control group(P<0.005). Conclusion : Dexamethasone treatment results in an increase in the SP-A mRNA and SP-A protein levels, suggesting that the pretranslational events in vivo may in part contribute to this process.

Resveratrol Ameliorates High-fat-induced Metabolic Complications by Changing the Expression of Inflammasome Markers and Macrophage M1 and M2 Markers in Obese Mice (라스베라트롤 투여가 고지방식이 비만쥐의 지방조직에서의 inflammasome과 대식세포 마커에 미치는 영향)

  • Lee, Young-Ran;Pitriani, Pipit;Park, Hee-Geun;Lee, Wang-Lok
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1462-1469
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    • 2017
  • The purpose of this study was to investigate the effects of resveratrol supplementation on inflammasome, inflammation, and macrophage markers in subcutaneous adipose tissue of high-fat-diet-induced obese mice. C57BL/6 mice were randomly assigned to three groups: normal diet control (NC; n=10), high-fat diet control (HC; n=10), or high fat with resveratrol (HRE; n=10) group. The mice were fed a high-fat diet (60% of calories from fat) or normal diet (18% of calories from fat). Resveratrol dissolved in a 0.1ml solution of dimethyl sulfoxide was supplemented orally at 25 mg/kg body weight. After 15 weeks, the body weight was significantly higher in the high-fat diet group than in the normal diet group. The inflammasome markers NLRP3, ASC, and caspase1 were significantly lower in the HRE group than in the HC group. The levels of an inflammation marker, IL-18, were also significantly lower in the HRE group than in the NC and HC groups. The levels of macrophage markers F480 and CD86 were significantly lower in the HRE group than in the HC group. The levels of the M2 macrophage marker CD206 were significantly decreased in the HC and HRE groups. Resveratrol had a positive effect on ameliorating the complications of high fat diet-induced obesity by reducing inflammasome and M1 macrophage gene expressions. However, resveratrol supplementation did not reduce inflammation gene expression.

Expression of mRNA for matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gingival and periodontal ligament fibroblasts treated with lipopolysaccharide from Prevotella intermedia (Prevotella intermedia의 세균내독소가 치은섬유아세포와 치주인대세포에서의 matrix metalloproteinase 및 tissue inhibitor of metalloproteinase의 발현에 미치는 영향)

  • Kim, Sung-Jo;Choi, Eun-Young;Choi, In-Soon;Lee, Ju-Youn;Choi, Jeom-Il;Kim, Chong-Kwan
    • Journal of Periodontal and Implant Science
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    • v.35 no.1
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    • pp.21-30
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    • 2005
  • Matrix metalloproteinases (MMPs) are a family of host-derived proteolytic enzymes and implicated in the remodeling and degradation of extracellular matrix under both physiological and pathological conditions. Connective tissue degradation in periodontal diseases is thought to be due to excessive MMP activities over their specific inhibitors. The effects of lipopolysaccharide (LPS) from Prevotella intermedia, one of the major putative pathogens of periodontitis, on the expression of mRNA for MMPs and tissue inhibitors of metalloproteinases (TIMPs) in human gingival and periodontal ligament fibroblasts were examined by reverse transcriptase-polymerase chain reaction (RT-PCR). The expression of mRNAs encoding MMP-1, -2, -3, -10, and -14 was increased in human gingival fibroblasts treated with p. intermedia LPS, whereas MMP-11 and TIMP-2 mRNA expression was decreased in these cells stimulated with LPS. P. intermedia LPS increased the MMP-1, -2, -10, -11, and -14 mRNA expression and decreased TIMP-1 and -2 mRNA expression in human periodontal ligament fibroblasts. These findings imply that P. intermedia LPS may play an important role in the connective tissue degradation in periodontitis.