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The Effect of Hydrogen Peroxide on Inducible Nitric Oxide Synthase Expression in Murine Macrophage RA W264.7 Cells (Murine macrophage RAW264.7에서 과산화수소가 유발형 산화질소 합성효소의 발현에 미치는 영향)

  • Ahn, Joong-Hyun;Song, Jeong-Sup
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.2
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    • pp.172-183
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    • 1999
  • Background: Nitric oxide is a short-lived effector molecule derived from L-arginine by the nitric oxide synthase(NOS). Nitric oxide plays a role in a number of physiologic and pathophysiologic functions including host defense, edema formation, and regulation of smooth muscle tone. Some kinds of cells including macrophage are known to produce large quantities of nitric oxide in response to inflammatory stimuli such as interleukin-$1\beta$(IL-$1\beta$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), interferon-$\gamma$(IFN-$\gamma$) and lipopolysaccharide(LPS). Reactive oxygen species are also known to be important in the pathogenesis of acute cell and tissue injury such as acute lung injury model Methods: Using the RA W264.7 cells, we have examined the ability of oxidant hydrogen peroxide($H_2O_2$) to stimulate nitric oxide production and inducible NOS mRNA expression. Also, we have examined the effects of NOS inhibitors and antioxidants on $H_2O_2$ induced nitric oxide production. Results: Stimulation of RAW264.7 cells with combinations of 100 ng/ml IL-$1\beta$, 100 ng/ml TNF-$\alpha$, and 100 U/ml IFN-$\gamma$ or 100 U/ml IFN-$\gamma$ and $1{\mu}g/ml$ LPS induced the synthesis of nitric oxide as measured by the oxidation products nitrite($NO_2^-$) and nitrate($NO_3^-$). Addition of $250 {\mu}M-2$ mM $H_2O_2$ to the cytokines significantly augmented the synthesis of $NO_2^-$ and $NO_3^-$(p<0.05). When cells were incubated with increasing concentrations of $H_2O_2$ in the presence of IL-$1\beta$, TNF-$\alpha$ and IFN-$\gamma$ at constant level, the synthesis of $NO_2^-$ and $NO_3^-$ was dose-dependently increased(p<0.05). $N^G$-nitro-L-arginine methyl ester(L-NAME), dose dependently, significantly inhibited the formation of $NO_2^-$ and $NO_3^-$ in cells stimulated with LPS, IFN-$\gamma$ and $H_2O_2$ at constant level(p<0.05). Catalase significantly inhibited the $H_2O_2$-induced augmentation of cytokine-induced $NO_2^-$ and $NO_3^-$ formation(p<0.05). But, boiled catalase did not produce a significant inhibition in comparison with the native enzyme. Another antioxidant 2-mercaptoethanol and orthophenanthroline dose-dependently suppressed $NO_2^-$ and $NO_3^-$ synthesis(p<0.05). Northern blotting demonstrated that H:02 synergistically stimulated the cytokine-induced iNOS mRNA expression in RA W264.7. Conclusion: These results suggest that $H_2O_2$ contributes to inflammatory process by augmenting the iNOS expression and nitric oxide synthesis induced by cytokines.

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Removal of Alkali Metal Ion using Inorganic Ion Exchanger (무기이온교환제를 이용한 알카리 금속이온 제거)

  • Ha, Ji-Won;Yi, Kwang Bok;Lee, Si Hyun;Rhee, Young-Woo;Kim, Jong-Nam
    • Korean Chemical Engineering Research
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    • v.46 no.2
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    • pp.423-429
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    • 2008
  • Currently, Ash-free clean coal producing process by solvent extraction is under development. The produced ash-free clean coal can be directly combusted in a gas turbine which results in substantial improvement of power generation efficiency. However, the clean coal produced by the solvent extraction still contain trace amount of alkali metal which may cause corrosion on turbine blades during the direct combustion. In present work ${\alpha},{\beta}$-metal (Zr and Ti) phosphates and H-Y zeolite were synthesized and their ion exchange characterizations were investigated for the application on alkali metal removal for clean coal production. $Na^+$ ion removal capacities of the metal phosphates and H-Y zeolite were measured and compared in both aqueous solution (100 ppmw, $Na^+$) and coal dissolved N-methyl-2-pyrrolidone (NMP, 12 ppmw $Na^+$) at elevated temperature. In aqueous solution, the ${\beta}$ form metal phosphates showed very high ion exchange capacities compared to ${\alpha}$ form. ${\beta}$ form metal phosphates also showed higher $Na^+$ removal capacities than H-Y zeolite. In ion exchange medium of NMP, all the ${\alpha}$ form metal phosphates showed over 90% of $Na^+$ ion removal efficiency in the temperature range of 200 to 400 while that of H-Y zeolite decreased as a half when the temperature was over 350. In addition, the regenerated metal phosphates by acid treatment showed no sign of degradation in $Na^+$ removal efficiency. Among the metal phosphates used, $Zr_{0.75}Ti_{0.25}(HPO_4)_2$ showed the best performance in $Na^+$ removal and is expected to be the most suitable inorganic ion exchanger for the alkali metal removal process.

Effect of Volatile Matter and Oxygen Concentration on Tar and Soot Yield Depending on Coal Type in a Laminar Flow Reactor (LFR에서 탄종에 따른 휘발분과 산소농도가 타르와 수트의 발생률에 미치는 영향)

  • Jeong, Tae Yong;Kim, Yong Gyun;Kim, Jin Ho;Lee, Byoung Hwa;Song, Ju Hun;Jeon, Chung Hwan
    • Korean Chemical Engineering Research
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    • v.50 no.6
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    • pp.1034-1042
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    • 2012
  • This study was performed by using an LFR (laminar flow reactor), which can be used to carry out different types of research on coal. In this study, an LFR was used to analyze coal flames, tar and soot yields, and structures of chars for two coals depending on their volatile content. The results show that the volatile content and oxygen concentration have a significant effect on the length and width of the soot cloud and that the length and width of the cloud under combustion conditions are less than those under a pyrolysis atmosphere. At sampling heights until 50 mm, the tar and soot yields of Berau (sub-bituminous) coal, which contains a large amount of volatile matter, are less than those of Glencore A.P. (bituminous) coal because tar is oxidized by the intrinsic oxygen component of coal and by radicals such as OH-. On the other hand, at sampling heights above 50 mm, the tar and soot yields of Berau coal are higher than those of Glencore A.P. coal by reacted residual volatile matter, tar and light gas in char and flame. With above results, it is confirmed that the volatile matter content and the intrinsic oxygen component in a coal are significant parameters for length and width of the soot cloud and yields of the soot. In addition, the B.E.T. results and the images of samples (SEM) obtained from the particle separation system of the sampling probe support the above results pertaining to the yields; the results also confirm the pore development on the char surface caused by devolatilization.

Development of RGD peptides grafted onto chitosan surfaces; Osteoblast interactions (RGD 펩타이드로 표면개질된 키토산막의 생물학적 영향)

  • Lee, Chang-Kyun;Hwang, Jeong-Hyo;Lee, Yong-Moo;Ku, Young;Rhyu, In-Chul;Lee, Seung-Jin;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.33 no.1
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    • pp.27-35
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    • 2003
  • 1. 목적 생체재료의 생체친화성을 증진시키고 치유를 촉진하기 위한 목적으로 생체재료의 생화학적 표면개질에 관한 연구가 널리 진행되고 있다. 이와 같은 목적으로 이용되어 온 부착분자에는 아미노산, 펩타이드, 단백질, 효소 및 성장인자들을 들 수 있으며, 이들 분자들을 금속, 골대체물질 및 폴리머와 같은 생체재료의 표면개질에 이용하여 왔다. 이 연구의 목적은 생체적합성이 우수하고 생분해성을 지닌 키토산으로 얇은 막을 제작한 후, 세포외 기질의 구성성분 중 세포부착에 관여하는 RGD 펩타이드를 부착시킨, 표면개질 키토산막의 생물학적 영향을 MG-63 조골양세포를 이용하여 관찰하는 것이다. 2. 방법 2% acetic acid에 키토산 가루를 녹여 만든 2% 키토산 용액으로 24-well 배양접시의 표면을 도포 후 24시간 동안 건조시켜 키토산막을 제작하였다. GRGDS 펩타이드를 cross-linker(EDC, NHS) (Sigma, MO, USA) 용액과 반응시켜서 펩타이드의 카르복실기를 활성화시켰다. 이들을 PBS 완충용액으로 수화시킨 키토산막과 결합시켜 펩타이드의 활성화된 카르복실기와 키토산의 아민기 간에 안정적인 아미드 결합(amide bond)이 형성되도록 하였다. 하루 동안 반응을 일으킨 후 PBS 완충용액과 증류수로 씻어내고 냉동 건조시킴으로써 GRGDS가 결합된 키토산막을 제작하였다. 재료 표면의 화학 성분을 알아보는데 사용되는 방법의 일종인 X-ray photoelectron spectroscopy(XPS) 분석을 통하여 부착분자가 키토산막에 결합된 여부를 확인하였다. GRGDS 펩타이드에 요오드를 결합시킨 후, 이것을 키토산막에 공유 결합시키고 XPS를 통해 요오드가 재료 표면에서 검출되는지를 검사하였다. 요오드가 검출된다면 이것은 키토산막 표면에 실제로 GRGDS 펩타이드가 존재하는 것을 의미하게 된다. 표면개질된 키토산막에 사람조골양세포인 MG-63을 접종하여 이를 실험군으로 하였고, 표면이 개질 되지 않은 키토산막을 대조군으로 하였다. 세포부착의 최적화 농도를 확인하기 위하여 GRGDS를 0.01, 0.05, 0.1, 0.25, 0.5, 1.0mg/ml의 농도로 준비하였다. 배양 후 1일, 7일째에 각 well에서 trypsin EDTA를 이용하여 세포를 분리한 후, 이를 원심 분리하여 세포수측정기를 이용하여 부착 세포의 수를 측정하여 세포의 부착 정도를 비교하였다. 배양 2시간, 24시간 후 주사전자현미경을 이용하여 키토산막에 부착된 세포의 양상을 관찰하였다. 3. 결과 XPS를 통한 표면의 화학 성분 분석 결과 GRGDS 펩타이드를 결합시킨 키토산막에서 요오드가 검출되었으며 펩타이드를 부착하지 않은 대조군에서는 검출되지 않았다. 따라서 cross-linker를 이용한 펩타이드와 키토산막의 공유결합을 확인할 수 있었다. 세포 배양 후 1일째 부착된 세포 수를 측정한 결과 0.1mg/ml 이상의 GRGDS 펩타이드 농도로 공유 결합시킨 키토산막에서 부착 세포 수가 다른 농도에 비해 유의성 있게 많이 관찰되었다. 이 농도 이하에서는 대조군과 실험군간에 세포부착의 유의한 차이가 없었다. 따라서 주사전자현미경을 이용한 부착 세포의 양상에 관한 관찰은 0.1mg/ml 농도의 펩타이드를 이용하였다. 세포 배양 7일째, 부착된 세포 수 측정 결과 GRGDS의 농도에 따른 유의성 있는 차이가 없었으며, 실험군과 대조군간에도 유의성 있는 차이가 없었다. 주사전자현미경 관찰결과 2시간 및 24시간 배양된 실험군 모두에서 별모양의 세포들이 키토산막 표면에 편평하게 잘 부착되어 있으며 많은 위족이 발달된 소견을 보인 반면, 대조군에서는 원형 또는 다각형 모양의 세포들이 실험군에 비해 부착이 덜 되어있는 양상을 보였다. 이 연구를 통하여 기능성 펩타이드를 생체재료의 표면에 공유결합 시키는 방법을 확립할 수 있었으며, RGD 펩타이드의 공유결합으로 표면개질된 키토산막이 조골세포의 부착능을 증진시킬 수 있음을 확인하였다. 표면개질된 생체재료를 소, 중동물에 적용시켜 생체 내에서의 생물학적 영향을 평가할 필요가 있으며, 이 실험의 결과는 향후 다양한 기능성 부착분자를 선발, 고안하여 임플란트용 생체재료의 표면개질에 이용하는 이른바 모방생체재료분야에 널리 활용될 수 있을 것으로 생각된다.

Antioxidant and Tyrosinase Inhibitory Effects of Brassica oleracea L. Fractions (적채 분획물의 항산화 및 Tyrosinase 억제 효과)

  • 윤경아;박윤자;배송자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.7-15
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    • 2004
  • This research was designed to investigate the antioxidant activity of red cabbage (Brassica oleracea L., BO) fractions on the liposomes consisted of L-$\alpha$-dilinoleoylphosphatidylcholine (DLPC-liposome) and the tyrosinase activity of BO fractions. The methanol extract of BO (BOM) was fractionated into five different partition layers: hexane (BOMH), ethylether (BOMEE), ethylacetate (BOMEA), butanol (BOMB) and aqueous (BOMA) layers. The antioxidant activities of BOM fractions toward oxidized DLPC-liposome were examined by spectrophotometry measuring oxidized conjugated dines. The antioxidant activities of BOMEE and BOMEA fractions toward oxidized DLPC-liposomes were similar to the antioxidant activities of $\alpha$-tocopherol and weaker than that of BHT. The synergy effects of antioxidation of BOMEE and BOMEA fractions added with vitamin C and $\alpha$-tocopherol had even stronger antioxidant activities than the fractions without vitamin C and $\alpha$-tocopherol. These results showed that the fractions of BOMEE and BOMEA could be developed as a potent antioxidant. Out of five different partition layers of BOM fractions, BOMEA exhibited the strongest tyrosinase activity of 94% at a concentration of 40 $\mu\textrm{g}$/mL. This result suggests BOMEA fractions inhibit the foramation of melanin and therefore can be used as the inhibitor of melanin synthesis. Results of antioxidant activity and tyrosinase inhibition indicate that useful bioactive substances exist in BOMEE and BOMEA fractions. Both fractions from BO (red cabbage) have the potential of being developed into health related products.

Neuroprotective Effects of Cirsium setidens, Pleurospermum kamtschaticumin, and Allium victorials Based on Antioxidant and p38 Phosphorylation Inhibitory Activities in SK-N-SH Neuronal Cells (SK-N-SH 신경세포내 항산화 효과와 p38 인산화 억제에 의한 곤드레, 누룩치 그리고 산마늘의 신경 보호 효과)

  • Chung, Mi Ja;Park, Yong Il;Kwon, Ki Han
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.3
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    • pp.347-355
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    • 2015
  • Oxidative stress is one of the key mechanisms involved in neuronal damage. Neuroprotective effects and underlying mechanisms of action of several wild vegetables, Cirsium setidens (CS), Pleurospermum kamtschaticumin (PK), and Allium victorials (AV), against oxidative stress induced by hydrogen peroxide in SK-N-SH cells were investigated. CS and AV up to $400{\mu}g/mL$ showed no detectable effects on cell viability of human SK-N-SH neuro-blastoma cells compared with control. Incubation of SK-N-SH cells with hydrogen peroxide resulted in significant induction of cell death and reaction oxygen species (ROS) production, whereas treatment of cells with CS and AV significantly reduced cell death and ROS production, respectively. Among the wild vegetables tested, CS and PK showed more effective DPPH radical scavenging activity than AV, whereas PK showed strong cytotoxicity in SK-N-SH cells compared with the control. CS showed much higher inhibitory effects on cell death and ROS generation against oxidative stress than AV. Thus, CS was selected for subsequent experiments. Ethyl acetate (EA), hexane, butanol, aqueous, and chloroform extracts from CS significantly inhibited cell death and ROS generation in SK-N-SH cells induced by oxidative stress. EA extract from CS (CS-EA) showed the highest DPPH radical-scavenging activity, intra-cellular ROS-scavenging activity, and neuroprotective effects. CS-EA attenuated apoptosis signal-regulating p38 activation by inhibiting phosphorylation. The findings suggest that CS-EA protects neuronal cells through antioxidant activity and inhibition of phosphorylation of p38 in brain neural cells.

Gender Roles, Accessibility, and Gendered Spatiality (성역할, 접근성, 그리고 젠더화된 공간성)

  • Kim, Hyun-Mi
    • Journal of the Korean Geographical Society
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    • v.42 no.5
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    • pp.808-834
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    • 2007
  • This study attempts to elucidate manifold dimensions of gendered accessibility experiences. How gender roles(household responsibilities) differentiate accessibility experiences between women and men is explored through the comparison of married dual-earner couples' parental status, using the US Portland activity-travel diary dataset with GIS-based geocomputation results of(time-geography based) space-time accessibility. First, this study shows how gender division of labor within the household still permeates current society, despite the widespread belief of the social change toward a gender-egalitarian society. Then, the study pays special attention to the way gender roles structure individual accessibility experiences of women and men differently, and, in turn, the way such accessibility experiences take a form of gendered spatiality. Gendered spatiality is examined through the analysis of accessibility space as well as activity space in order to ascertain women's home-attached and spatially entrapped characteristics. More household responsibilities throughout a day and, even more, the time constraint of picking up children at the daycare centers after work lead women's possible activity space to be more home-centered. The analysis of the spatio-temporal context of accessibility space makes gendered spatiality visible. However, the findings suggest that behavioral outcomes should be understood with an explicit awareness of constraints individuals face. It is because the revealed activity spaces can be not only an outcome of constraint but also an outcome of choice. Behavioral outcomes should not be treated as a straightforward expression of the level of constraints. It is problematic to expect that behavioral outcomes directly mirror the level of constraints. It is also problematic to suppose that the level of constraints can be straightforwardly elicited from revealed behavioral outcomes.

Validation of Satellite SMAP Sea Surface Salinity using Ieodo Ocean Research Station Data (이어도 해양과학기지 자료를 활용한 SMAP 인공위성 염분 검증)

  • Park, Jae-Jin;Park, Kyung-Ae;Kim, Hee-Young;Lee, Eunil;Byun, Do-Seong;Jeong, Kwang-Yeong
    • Journal of the Korean earth science society
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    • v.41 no.5
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    • pp.469-477
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    • 2020
  • Salinity is not only an important variable that determines the density of the ocean but also one of the main parameters representing the global water cycle. Ocean salinity observations have been mainly conducted using ships, Argo floats, and buoys. Since the first satellite salinity was launched in 2009, it is also possible to observe sea surface salinity in the global ocean using satellite salinity data. However, the satellite salinity data contain various errors, it is necessary to validate its accuracy before applying it as research data. In this study, the salinity accuracy between the Soil Moisture Active Passive (SMAP) satellite salinity data and the in-situ salinity data provided by the Ieodo ocean research station was evaluated, and the error characteristics were analyzed from April 2015 to August 2020. As a result, a total of 314 match-up points were produced, and the root mean square error (RMSE) and mean bias of salinity were 1.79 and 0.91 psu, respectively. Overall, the satellite salinity was overestimated compare to the in-situ salinity. Satellite salinity is dependent on various marine environmental factors such as season, sea surface temperature (SST), and wind speed. In summer, the difference between the satellite salinity and the in-situ salinity was less than 0.18 psu. This means that the accuracy of satellite salinity increases at high SST rather than at low SST. This accuracy was affected by the sensitivity of the sensor. Likewise, the error was reduced at wind speeds greater than 5 m s-1. This study suggests that satellite-derived salinity data should be used in coastal areas for limited use by checking if they are suitable for specific research purposes.

Field Application Techniques of Simultaneous Mating Disruptor Against Grapholita molesta and G. dimorpha (복숭아순나방과 복숭아순나방붙이에 대한 동시 교미교란제의 현장 적용 기술)

  • Cho, Jum-Rae;Park, Chang-Gyu;Park, Il-Kweon;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.57 no.3
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    • pp.209-220
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    • 2018
  • Mating disruption (MD) has been widely used to effectively control Grapholita molesta in apple orchards. A simultaneous mating disruption (SMD) techniques have been developed to control both G. molesta and G. dimorpha. This study was performed to determine the practical parameters to apply the SMD technique to field conditions. To determine the application amount of SMD lures, a dispenser containing 10 mg pheromone was placed at different numbers of trees in an orchard. Application at every other tree (= one dispenser per two trees) was relatively safe to expect effective MD efficiency in both wax and polyethylene (PE) formulations. One time application at the end of March was enough to maintain a year round MD efficacy against both species. A fence treatment using food trap was applied to prevent any immigratory mated females from nearby untreated regions. To enhance the food trap by adding host-derived secondary compounds, terpinyl acetate (TA) was screened to be effective to attract females of Grapholita molesta among six compounds contained in apple fruit extracts. Among different TA concentrations, 0.05% TA treatment was the most effective to attract the adults. A mixture of TA and sugar was effective to attract and kill females and called FAKT (female attract-to-kill techniques). FAKT was treated at approximately 6 m interval at the edge of the apple orchards. The females trapped by the FAKT included mated females possessing vitellogenic oocytes. SMD supplemented with FAKT maintained the high MD efficacy and significantly suppressed leaf damage induced by the two insect pests compared to control or single SMD treatment.

Apoptotic Effect of co-treatment with HS-1200 and Cisplatin on SCC25 Human Tongue Squamous Cell Carcinoma Cell Line (HS-1200과 cisplatin의 병용처리가 사람구강암세포에 미치는 세포자멸사 효과에 대한 연구)

  • Kim, Duk-Han;Kim, In-Ryoung;Park, Bong-Soo;Ahn, Yong-Woo;Jeong, Sung-Hee
    • Journal of Oral Medicine and Pain
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    • v.38 no.3
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    • pp.221-233
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    • 2013
  • Bile acids are polar derivatives of cholesterol essential for the absorption of dietary lipids and regulate the transcription of genes that control cholesterol homeostasis. Recently it have been identified the synthetic chenodeoxycholic acid (CDCA) derivatives HS-1200 and cisplatin showed apoptisis-inducing activity on various cancer cells in vivo and in vitro. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with HS-1200 and cisplatin on human tongue squamous cell carcinoma cells (SCC25 cells). To investigate whether the co-treatment with HS-1200 and cisplatin compared to each single treatment efficiently reduces the viability of SCC25 cells, MTT assay was conducted. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining and an analysis DNA hypoploidy. Westen blot analysis and immunofluorescent staining were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following this co-treatment. Furthermore, proteasome activity and mitochondrial membrane potential (MMP) change were also assayed. In this study, co-treatment with HS-1200 and cisplatin on SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensations, DNA fragmentation, reduction of MMP and proteasome activity, the increase of Bax and the decrease of Bcl-2, decrease of DNA content, the release of cytochrome c into cytosol, translocation of AIF and DFF40 (CAD) onto nuclei, and activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD) whereas each single treated SCC25 cells did not show these patterns. Although the single treatment of $25{\mu}M$ HS-1200 and $4{\mu}g/ml$ cisplatin for 24 h did not induce apoptosis, the co-treatment of these reagents prominently induced apoptosis. Therefore our data provide the possibility that the combination therapy with HS-1200 and cisplatin could be considered as a novel therapeutic strategy for human squamous cell carcinoma.