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C6 glia 세포에서 유도성 Nitric Oxide Synthase 유전자 발현조절에 관한 연구
배진영,허강민,배소현,박지선,이충재,이재흔,석정호 충남대학교 의과대학 의학연구소 2003 충남의대잡지 Vol.30 No.1
To investigate transcriptional regulation of iNOS gene by LPS and cytokines, the production of NO, expression of iNOS mRNA and protein, binding activity of nuclear factor-kappa B(NF-kB), and promoter activity of iNOS gene were examined in rat C6 glial cells. LPS, interferon-gamma(IFN-γ), and tumor necrosis factor-alpha (TNF-α) stimulated the production of NO, which was increased synergistically by co-treatment. By the treatment of LPS, iNOS mRNA expression was initiated at 1 h, markedly increased by 3 h, and decreased gradually afterward. iNOS mRNA expression was markedly enhanced by mixture of LPS, IFN-γ and TNF-α. iNOS protein synthesis was increased by the treatment of mixture LPS and cytokine mixture. Treatment of LPS stimulated NF-kB activation, and the activation reached to the maximum level at 30 min, and the treatment of mixture of LPS and cytokines increased the activation. To determine the effect of NF-kB binding activity on iNOS promoter activation, CAT assay was performed. iNOS promoter activity was increased by the treatment with LPS for 5.5 h, and further increased by the combined treatment with LPS and cytokines. These results suggest that NF-kB activation by LPS and cytokines may play a significant role in the induction of the iNOS gene.
골격근 근형질세망의 ATP유도 ^45Ca-uptake에 대한 Thapsigargin및 Vanillylnonanamide의 영향
황의강,배소현,홍장희,허강민,김진회,이재흔,석정호 충남대학교 의과대학 지역사회의학연구소 1999 충남의대잡지 Vol.26 No.2
To investigate the effect of thapsigargin(THP) and vanillylnonanamide(VN), derivative of capsaicin, on the ATP-induced ^45Ca-uptake in the sarcoplasmic reticulum(SR) of the skeletal muscle, the SR vesicles were prepared from the back muscle of the rabbit, and ^45Ca-uptake was carried out. The results as follows: ATP-induced ^45Ca-uptake of skeletal muscular SR was significantly increased by 1 mM ATP. It was significantly blocked by 5 μM THP, but slightly decreased by 20 μM VN. The combined effect of THP and VN on the 45Ca-uptake of the SR vesicles was much potentiated than the sum of each effect of them. The above results suggest that the action of VN, being slightly influential to ATP-induced Ca-uptake but potentiating the effect of THP on the Ca-uptake, might be related with direct perturbation of the SR membrane or exposure of the THP-binding site.
신경교세포 및 RAW 264.7 세포에서 Protein kinase의 활성에 의한 유도성 Nitric oxide synthase의 발현
박상철,노삼길,배소현,박지선,이충재,허강민,석정호,이재흔 충남대학교 의학연구소 2003 충남의대잡지 Vol.30 No.2
NO(nitric oxide) plays an important role as neurotransmitter or cytokine, and pathologic factor for some diseases by the large amount production with iNOS(inducible NO synthase) expression in macrophages or glial cells. The expression of iNOS is regulated by various cytokines, protein kinases and transcription factors. In this experiment, to investigate the roles of progein kinase and NF-kB for iNOS expression, the effects of PMA(phorbol 12-myristate 13-acetate), cAMP, and various protein kinase inhibitors on LPS(lipopolysaccharide)-induced iNOS mRNAN expression and nuclear NF-kB binding complex were examined in C6 glial cells and RAW 264.7 cells. In C6 glial cells, iNOS mRNA expression by LPS was induced from 1 hour and peak at 3 hour after treatment. In RAW 264.7 cells, the mRNA was observed from 3 hour and peak at 6 hour. PMA enhanced markedly LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but did not much influence on LPS-induced iNOS mRNA expression in RAW 264.7 cells, in spite of increased LPS-induced NF-kB binding complex at 30 min. cAMP(dibutyryl cAMP) did not much influence on LPS-induced iNOS mRNA expression, by increased LPS-induced NF-kB binding complex in C6 glial cells. However, in RAW 264.7 cells, cAMP increased slightly LPS-induced iNOS mRNA expression without change of NF-kB binding complex. Staurosporine did not influence on LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased LPS-induced iNOS mRNA expression and NF-kB binding complex. Ro-31-8220 did not much influence on LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased significantly LPS-induced iNOS mRNA expression in spite of increased LPS-induced NF-kB binding complex for 3hours. G 6976 did not much influence on LPS-induced iNOS mRNA expression with decreased NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased iNOS mRNA expression without influence on LPS-induced NF-kB binding complex. Genistein did not influence on LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased LPS-induced iNOS mRNA expression inspite of increased NF-kB binding complex. These results suggest that LPS-induced regulation of iNOS expression or NF-kB activity in C6 glial cells, might be different from RAW 264.7 cells through various protein kinases or other factors.
Effect of Altered Blood Coagulability on Tumor Metastasis
Gang, Seok Jin,Kim, Sun Moo CATHOLIC MEDICAL CENTER 1986 Bulletin of the Clinical Research Institute Vol.14 No.1
The effect of hypocoagulabflity and hypercoagulability on the metastases was studied in female A/J inbred mice on which intravenous infusion had been performed with 4×10exp(4)/0.1 ml SCK mammary carcinoma cells. The mice were divided into six groups-control, heparin-injected, prednisolone-injected, fibrinogen-injected, factor XIII-injected and fibrinogen plus factor XIII-injected. The mice were sacrificed 3 days (early phase of tumor metastasis) and 21 days (late phase of tumor metastasis) after tumor cell infusion. Histological and immunofluorescent observations were made on lungs. In colony assay in vivo, lung colonies were counted when mice were sacrificed 21 days after tumor cell infusion. The results were as follows: 1. The control group revealed occasional tumor clusters and colonies. Fibrin coated tumor cells were frequently shown. 2. The heparin-injected group revealed rare tumor colonies showing a weak fluorescence. 3. The other four groups (prednisolone, fibrinogen, factor XIII, and fibrinogen plus factor XIII-injected) showed numerous tumor clusters and colonies showing a strong fluorescence.