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.

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.

일차배양된 설치류 호흡기 상피세포로부터의 점액소 분비에 대한 수종 약물의 영향

이충재,석정호,이재흔,허강민,박지선,배소현,노삼길,박상철 충남대학교 의학연구소 2003 충남의대잡지 Vol.30 No.2

1. PKC activator인 PMA는 일차배양 HTSE세포로부터의 뮤신분비를 0.1μM 농도에서 30%, 1μM 농도에서 80% 가량 증가시켰다. 2. 식물 유래 성분으로, flavonoid의 일종인 TFR은 일치배양 HTSE 세포로부터의 뮤신분비를 10μM 농도에서 50%, 100μM 농도에서 80% 가량 증가시켰다. 3. 양이온성 폴리펩티드인 PLL 및 PLA는 일차 배양 HTSE 세포로부터의 뮤신분비를 0.01 - 10μM 농도에서 용량의존적으로 감소시켰다. 4. 결론적으로, 본 연구에서 얻어진 결과들은 새로운 거담제 및 점액용해제나 단백분해 효소제가 아닌 호흡기 류신의 생성/분비를 조절해 줄 수 있는 신개념의 약물을 개발함에 있어 극히 일부분이나마 단서를 제공하고 있다고 하루 수 있을 것이다. In the present study, we tried to investigate whether phorbol myristate acetate(PMA), trihydroxymethoxy-flavanone rutinoside(TFR) and cationic polypeptides significantly affect mucin release(secretion) from cultured hamster tracheal surface epithelial cells. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24 hr and chased for 30 min in the presence of each agent to assess the effect on 3H-mucin release. The results were as follows : (1) Both PMA and TFR significantly increased mucin release from cultured HTSE cells ; (2) Cationic polypeptides including po1y-L-lysine(PLL, mw 7,500) and poly-L-arginine(PLA, mw 10,800) significantly inhibited mucin release from cultured HTSE cells, in a dose-dependent manner. This finding suggests us that PMA and TFR be further studied for the possible use as mild expectorants and cationic polypeptides might function as a regulator for hyper-secretion of mucus, both by direct acting on airway mucin-secreting cells, during the treatment of chronic airway diseases.

논토양중 카드뮴 유효도와 수도의 흡수이행에 미치는 석회 및 Humic acid 사용효과

김민경,김원일,정구복,박광래,윤순강,엄기철 한국환경농학회 2004 한국환경농학회지 Vol.23 No.1

토양중 중금속이 식물에 흡수 및 이행되는 것을 경감시켜 여기에서 생산된 농산물의 안전성 향상을 위한 기초자료를 확보하고 논토양의 카드뮴 유효도와 흡수이행에 미치는 석회와 humic acid 처리 효과를 구명하기 위하여 실내경험과 포트실험을 수행하였다. 실내실험에서 공시토양에 2.5와 5.0 ton/ha의 석회와 1%와 2% humic acid를 처리한 후 토양중 가용성 카드뮴 함량 변화를 조사한 결과, 두 처리 모두 14일까지는 담수상태에서 처리효과로 인하여 가용성 카드뮴 함량이 감소하다가 그 이후에는 토양의 완충능에 의해 다시 증가하는 경향이었다. 이때, 토양의 가용성 카드뮴 함량은 토양 pH 및 양이온치환용량과 부의 상관이 있었다. 2.5 ton/ha의 석회와 1% 의 humic acid를 처리한 후 담수시켜 안정화된 다음 벼를 재배하여 조사한 결과, 석회와 humic acid 처리에 비해 분얼기와 수확기의 치화태 및 수용태 카드뮴 함량이 낮아졌으나, 분얼기에 비해 수확기에 토양중 이동이 어려운 산화물 및 탄산염태와 황화물 및 잔류태의 함량이 증가하였다. 식물에 흡수가 용이한 형태로 알려진 치환태와 수용태는 모두 토양 pH와 고도의 부의 상관이 있었다. 수확기의 줄기, 잎 및 현미 건물중은 석회 및 humic acide 처리구에서 대조구에 비해 모두 높았고, 특히 석회 처리구의 건물중이 현저하게 높았다. 석회 처리구에서 줄기와 잎의 카드뮴 함량은 각각 1.01과 0.37㎎/㎏으로 대조구와 비슷하였으나, 뿌리나 현미의 카드뮴 ?량은 각각 2.11과 0.09㎎/㎏으로 대조구에 비해 낮았다. 지상부와 뿌리의 카드뮴 함량은 humic acid 처리에 의해 현저히 낮아졌으며, 특히 현미로의 카드뮴 이행도 월등히 낮게 나타난다. This study was conducted to know the effect of lime and humic acid on cadmium availability and its uptake by plant grown in contaminated paddy soils with heavy metal. The treatment levels of lime were 2.5 and 5.0 ton/ha and that of humic acid were 1 and 2%. The contents of 0.1 N HCl extractable Cd were reduced with lime and humic acid and were negatively correlated with CEC as well as soil pH. The sequential extraction procedure was used to fractionate the heavy metals in soil into the designated from exchangeable (0.5 M KNO₃), water soluble (H₂O), organically bound (0.5 M NaOH), carbonate (0.05 M Na₂-EDTA) and sulfide/residual (4 M HNO₃). In soil amended with 2.5 ton/ha lime and 1% humic acid, chemical forms of Cd at tillering stage were predominant exchangeable+water soluble extractable Cd, whereas that at harvesting stage were predominant carbonate+sulfide/residual extractable Cd. The exchangeable forms of Cd in soil with lime and humic acid were negatively correlated with soil pH during the harvesting period. Total absorbed Cd of paddy rice tended to occur in the order of root > stem > leaf > brown rice. Cd con- tents of brown rice with lime and humic acid treatment were 0.09 and 0.08 mg/kg, respectively. That were lower than control, 0.20 ㎎/㎏. It could be that treatment of lime and humic acid in polluted soil by heavy metals would reduce the uptake of heavy metals by plants and be a temporary method of reclamation at the highly heavy metal contaminated soils.

Evaluation of residual tensile strength of fire-damaged concrete using a non-linear resonance vibration method

Park, Sun-Jong,Park, Gang-Kyu,Yim, Hong Jae,Kwak, Hyo-Gyoung Thomas Telford Ltd. 2015 Magazine of concrete research Vol.67 No.5

Effects of Proinflammatory Cytokines and Natural Products on Mucin Release from Cultured Hamster Tracheal Surface Epithelial Cells

Park, Ji-Sun,Kim, Hyoung-Soo,Seok, Jeong-Ho,Hur, Gang-Min,Park, Jong-Sun,Seo, Un-Kyo,Lee, Choong-Jae The Korean Society of Pharmacology 2004 The Korean Journal of Physiology & Pharmacology Vol.8 No.6

In this study, we investigated whether TNF-alpha, IL-1beta, CTMA (carboxymethyl trimethylammonium) and LPD (Lup-20[29]-ene-3beta,28-diol) affect mucin release from airway goblet cells and compared the activities of these agents with the inhibitory action of PLL and the stimulatory action of ATP on mucin release. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with $^3H-glucosamine$ for 24 h and chased for 30 min in the presence of varying concentrations of each agent to assess the effects on $^3H-mucin$ release. The results were as follows: TNF-alpha, CTMA and LPD increased mucin release at the highest concentration, but IL-1beta did not. We conclude that CTMA and LPD can stimulate mucin release by directly acting on airway mucin-secreting cells, and suggest that these agents should be further investigated for the possible use as mild expectorants during the treatment of chronic airway diseases.

Effect of Erythromycin on Basal and Stimulated Mucin Release from Cultured Hamster Tracheal Surface Epthelial Cells

( Ji Sun Park ),( Jeong Ho Seok ),( Gang Min Hur ),( Jae Heun Lee ),( Kyeung A Park ),( Choong Jae Lee ) 한국응용약물학회 2004 Biomolecules & Therapeutics(구 응용약물학회지) Vol.12 No.2

비선형 공진기법을 이용한 콘크리트의 화재 손상 영향인자 분석

박강규(Gang-Kyu Park),박선종(Sun-Jong Park),임홍재(Hong Jae Yim),곽효경(Hyo-Gyoung Kwak) 한국비파괴검사학회 2015 한국비파괴검사학회지 Vol.35 No.2

본 연구에서는 비선형 음향효과를 기반으로 한 비선형 공진기법을 도입하여 콘크리트의 배합비 및 화재 손상 조건(노출온도, 손상 후 경과기간)이 화재 손상 콘크리트에 미치는 영향을 파악하였다. 도입된 비선형 공진기법을 통해 기존 선형 탄성파 기반 평가 기법 대비 향상된 민감도를 나타내는 비선형인자를 측정하였으며, 쪼갬 인장강도 측정을 통해 배합비 및 화재 손상 조건에 따른 콘크리트의 잔존재료물성 평가를 수행하였다. 얻어진 결과를 토대로 배합비, 노출온도, 손상 후 경과기간이 화재 손상 콘크리트에 미치는 영향을 분석하였다. 추가적으로 쪼갬 인장강도비와 비선형인자의 직접적인 관계를 제시하였으며, 비선형 공진기법을 이용한 화재 손상 콘크리트의 잔존 강도 추정에 대한 가능성을 확인하였다. In this study, the effects of different mix proportions and fire scenarios (exposure temperatures and post-fire-curing periods) on fire-damaged concrete were analyzed using a nonlinear resonance vibration method based on nonlinear acoustics. The hysteretic nonlinearity parameter was obtained, which can sensitively reflect the damage level of fire-damaged concrete. In addition, a splitting tensile strength test was performed on each fire-damaged specimen to evaluate the residual property. Using the results, a prediction model for estimating the residual strength of fire-damaged concrete was proposed on the basis of the correlation between the hysteretic nonlinearity parameter and the ratio of splitting tensile strength.

Poster Session : Phorbol 12-myristate 13-acetate protects against necrotic cell death in response to death receptor-Ligation

( Hee Sun Byun ),( Kyeong Ah Park ),( Min Ho Won ),( Kuem Jin Yang ),( Sang Hee Shin ),( Long Zhen Piao ),( Jin Young Kwak ),( Ji Won Lee ),( Jong Sun Park ),( Jeong Ho Seok ),( Zheng Gang Liu ),( Gan 한국생화학분자생물학회 (구 한국생화학회) 2006 생화학분자생물학회 춘계학술발표논문집 Vol.2006 No.-

Pancreatic adenocarcinoma up-regulated factor (PAUF), a novel up-regulated secretory protein in pancreatic ductal adenocarcinoma.

Kim, Sun A,Lee, Yangsoon,Jung, Dawoon E,Park, Kyung Hwa,Park, Jeong Youp,Gang, Jingu,Jeon, Sun Bok,Park, Eui Chul,Kim, Young-Gun,Lee, Bogman,Liu, Qing,Zeng, Wen,Yeramilli, Subramanyam,Lee, Soojin,Koh, Japanese Cancer Association 2009 CANCER SCIENCE Vol.100 No.5

<P>The identification of novel tumor-specific proteins or antigens is of great importance for diagnostic and therapeutic applications in pancreatic cancer. Using oligonucleotide microarrays, we identified a broad spectrum of differentially expressed pancreatic cancer-related genes. Of these, we selected an overexpressed expressed sequence taq and cloned a 721-bp full-length cDNA with an open reading frame of 196 amino acids. This novel gene was localized on the Homo sapiens 16p13.3 chromosomal locus, and its nucleotide sequence matched the Homo sapiens similar to common salivary protein 1 (LOC124220). We named the gene pancreatic adenocarcinoma up-regulated factor. The pancreatic adenocarcinoma up-regulated factor was secreted into the culture medium of pancreatic adenocarcinoma up-regulated factor-overexpressing Chinese hamster ovary cells, had an apparent molecular mass of approximately 25 kDa, and was N-glycosylated. The induction of pancreatic adenocarcinoma up-regulated factor in Chinese hamster ovary cells increased cell proliferation, migration, and invasion ability in vitro. Subcutaneous injection of mice with Chinese hamster ovary/pancreatic adenocarcinoma up-regulated factor cells resulted in 3.8-fold greater tumor sizes compared to Chinese hamster ovary/mock cells. Reverse transcription-polymerase chain reaction and western blotting with antirecombinant human pancreatic adenocarcinoma up-regulated factor antibodies confirmed that pancreatic adenocarcinoma up-regulated factor was highly expressed in six of eight pancreatic cancer cell lines. Immunohistochemical staining of human pancreatic cancer tissues also showed pancreatic adenocarcinoma up-regulated factor overexpression in the cytoplasm of cancer cells. Transfection with pancreatic adenocarcinoma up-regulated factor-specific small-interfering RNA reduced cancer cell migration and invasion in vitro. Treatment with antirecombinant human pancreatic adenocarcinoma up-regulated factor in vitro and in vivo reduced proliferation, migration, invasion, and tumorigenic ability. Collectively, our results suggest that pancreatic adenocarcinoma up-regulated factor is a novel secretory protein involved in pancreatic cancer progression and might be a potential target for the treatment of pancreatic cancer.</P>