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( Yu Ah Hong ),( Keum Jin Yang ),( So Young Jung ),( Yoon Kyung Chang ),( Cheol Whee Park ),( Chul Woo Yang ),( Suk Young Kim ),( Hyeon Seok Hwang ) 대한신장학회 2017 Kidney Research and Clinical Practice Vol.36 No.2
Background: Vitamin D is considered to exert a protective effect on various renal diseases but its underlying molecular mechanism remains poorly understood. This study aimed to determine whether paricalcitol attenuates inflammation and apoptosis during lipopolysaccharide (LPS)-induced renal proximal tubular cell injury through the prostaglandin E<sub>2</sub> (PGE<sub>2</sub>) receptor EP4. Methods: Human renal tubular epithelial (HK-2) cells were pretreated with paricalcitol (2 ng/mL) for 1 hour and exposed to LPS (1 μg/mL). The effects of paricalcitol pretreatment in relation to an EP4 blockade using AH-23848 or EP4 small interfering RNA (siRNA) were investigated. Results: The expression of cyclooxygenase-2, PGE<sub>2</sub>, and EP4 were significantly increased in LPS-exposed HK-2 cells treated with paricalcitol compared with cells exposed to LPS only. Paricalcitol prevented cell death induced by LPS exposure, and the cotreatment of AH-23848 or EP4 siRNA offset these cell-protective effects. The phosphorylation and nuclear translocation of p65 nuclear factor-kappaB (NF-κB) were decreased and the phosphorylation of Akt was increased in LPS-exposed cells with paricalcitol treatment. AH-23848 or EP4 siRNA inhibited the suppressive effects of paricalcitol on p65 NF-κB nuclear translocation and the activation of Akt. The production of proinflammatory cytokines and the number of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive cells were attenuated by paricalcitol in LPS exposed HK-2 cells. The cotreatment with an EP4 antagonist abolished these anti-inflammatory and antiapoptotic effects. Conclusion: EP4 plays a pivotal role in anti-inflammatory and antiapoptotic effects through Akt and NF-κB signaling after paricalcitol pretreatment in LPS-induced renal proximal tubule cell injury.
12-O-Tetradecanoylphorbol-13-Acetate에 의한 HL-60 세포 분화유도중 Vimentin 유전자 전사조절에 대한 AP-1의 역할
임규,김진희,권도원,김승민,이명선,윤경아,손미영,박종일,윤완희,황병두 忠南大學校 癌共同硏究所 1998 癌共同硏究所 硏究誌 Vol.2 No.1
Purpose: To gain insight on the role of AP-1 in transcriptional regulation of vimentin gene during differentiation of HL-60 cells by 12-O-tetradecanoylphorbol-13-acetate (TPA), the levels of vimentin mRNA and AP-1 have been investigated with Northern blot hybridization and DNA mobility shift assay. Materials and Methods: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum and antibiotics in a humidified 5% CO_(2) at 37°C. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. EcoRI fragment of pVIM-GEM was used as probe for vimentin mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA probe(Upper strand, 5'-CGCTTGATGAGTCAGCCG- 3') for AP-1 binding activity was mixed with nuclear extracts in a 20 μl reaction volume containing 300 mM KC1, 60 mM HEPES, pH 7.9, 25 mM MgCl_(2), 1 mM EDTA, 1 mM DTT, 60% glycerol, and 2μg of poly[dI-dC]. Results: TPA increased vimentin mRNA levels, with maximal stimulation reached at 24 hr. The level of vimentin mRNA was induced in proportion to the concentration of TPA. TPA-induced vimentin mRNA was almost reduced by actinomycin-D pretreatment. TPA-induced stimulation of vimentin gene was completely reduced by staurosporin pretreatment. In DNA mobility shift assay, AP-1 newly appeared at 24 hr during TPA-induced differentiation and was almost not detected after the pretreatment of staurosporin. Conclusions: These results suggest that the induction of vimentin mRNA during TPA-dependent differentiation in HL-60 cells may be mediated by protein kinases C signal transduction and AP-1 is important to transcriptional regulation.
RAW 264.7 세포에 대한 중국산 천연 광물성 섬유 TAFMAG의 독성효과
임영,한진구,김지홍,김현욱,김은경,김경아,장황신 大韓産業醫學會 1999 대한직업환경의학회지 Vol.11 No.3
Objectives : This study was designed to evaluate cytotoxicity of TAFMAG, which is a trade name of natural mineral fiber mined and produced in China. Methods : The cytotoxicity of TAFMAG was evaluated by measuring iron content, lipid peroxidation, erythrocyte hemolysis, and cytotoxicity in vitro. These results were compared with the data of chrystotile and wollastonite as a positive and negative control, respectively. Results : There was significant increase of Fenton activity in TAFMAG and chrysotile with dose-response pattern. The iron chelating agent, desferrioxamine, significantly decreased Fenton activity of the particulates except wollastonite. TAFMAG and chrysotile fibers significantly increased malondialdehyde concentration from lipid peroxidation of the red blood cell membrane. In erythrocyte hemolysis test, TAFMAG & chrysotile had stronger effect on erythrocyte hemolysis than wollastonite with the concentration of 1,000g/ml. Furthermore, TAFMAG was more hemolytic than chrysotile with the concentration of 5,000g/ml. There was a significant cytotoxic effect in TAFMAG and chrysotile on RAW cell compared with wollastonite. Conclusions : In vitro study suggested that TAFMAG may have a similar health hazard as usual asbestos.
12-O-Tetradecanoylphorbol-13-Acetate에 의한 HL-60 세포 분화유도중 Vimentin 유전자 전사조절에 대한 AP-1의 역할
임규,김진희,권도원,김승민,이명선,윤경아,손미영,박종일,윤완희,황병두 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-
Purpose: To gain insight on the role of AP-1 in transcriptional regulation of vimentin gene during differentiation of HL-60 cells by 12-O-tetradecanoylphorbol-13-acetate (TPA), the levels of vimentin mRNA and AP-1 have been investigated with Northern blot hybridization and DNA mobility shift assay. Materials and Methods: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum and antibiotics in a humidified 5% CO_2 at 37℃. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. EcoRI fragment of pVIM-GEM was used as probe for vimentin mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA probe(Upper strand, 5'-CGCTTGATGAGTCAGCCG- 3') for AP-1 binding activity was mixed with nuclear extracts in a 20 μl reaction volume containing 300 mM KCI, 60 mM HEPES, pH 7.9, 25 mM MgCI_2, 1 mM EDTA, 1 mM DTT, 60% glycerol, and 2 ㎍ of poly[dI-dC]. Results: TPA increased vimentin mRNA levels, with maximal stimulation reached at 24 hr. The level of vimentin mRNA was induced in proportion to the concentration of TPA. TPA-induced vimentin mRNA was almost reduced by actinomycin-D pretreatment. TPA-induced stimulation of vimentin gene was completely reduced by staurosporin pretreatment. In DNA mobility shift assay, AP-1 newly appeared at 24 hr during TPA-induced differentiation and was almost not detected after the pretreatment of staurosporin. Conclusions: These results suggest that the induction of vimentin mRNA during TPA-dependent differentiation in HL-60 cells may be mediated by protein kinases C signal transduction and AP-1 is important to transcriptional regulation.
Hwang, Jin-Ah The Korean Nutrition Society 2009 Nutrition Research and Practice Vol.2 No.1
It has been reported that dietary polyunsaturated fats (PUFA) increase liver injury in response to ethanol feeding. We tested the hypothesis that diets rich in linoleic acid (18:2n-6) would affect acute liver injury after acetaminophen injection and that protein restriction might exacerbate the liver injury. We examined effects of feeding diets with either 15% (wt/wt) corn oil or 14% beef tallow and 1% corn oil for six weeks with either 6 or 20 g/100 g protein on acute hepatotoxicity. After the feeding period, liver injury was induced by injecting either with 600 mg/kg body weight acetaminophen suspended in gum arabic-based vehicle, or with vehicle alone during fasting status. Samples of liver and plasma were taken for analyses of hepatic glutathione (GSH) levels and liver-specific enzymes [(Glutamate-pyruvate transaminase (GPT) and glutamate-oxaloacetate transaminase (GOT)], respectively. Whereas GSH level was significantly lower in only group fed 15% corn oil with 6 g/100 g protein among acetaminophen-treated groups, activities of GPT and GOT were significantly elevated in all groups except the one fed beef tallow with 20 g/100 g protein, suggesting low protein might exacerbate drug-induced hepatotoxicity. The feeding regimens changed the ratio of 18:2n-6 to oleic acid (18:1n-9) in total liver lipids approximately five-fold, and produced modest changes in arachidonic acid (20:4n-6). We conclude that diets with high 18:2n-6 promote acetaminophen-induced liver injury compared to diets with more saturated fatty acids (SFA). In addition, protein restriction appeared to exacerbate the liver injury.
황진아(Jin-Ah Hwang),남기군(Gi-Gun Nam),이장명(Jang-Myung Lee) 대한전기학회 2006 정보 및 제어 심포지엄 논문집 Vol.2006 No.1
This paper proposes a method of container position measurement using automatic landing system that is estimated by a laser range finder. In the most of container position measurement methods, CCD cameras or laser scanners have been used to get the source data. However those sensors are not only weak for disturbances, for examples, the light, fog, and rain, but also the system cost is high. When the spreader arrives the goal position, it is still swung by inertia or by wind effect. In this paper, the spreader swung data have been used to find the container position. The laser range finder is equipped in the front side of spreader. It can measure distance and relative position between spreader and container. This laser range finder can be rotated as desired by a motor. And a tilt sensor is equipped on the spreader to measure spreader sway. We estimate the relative position information between the spreader and a container using the laser range finder and tilt sensor through the geometrical analysis.