Nutlin-3 induces HO-1 expression by activating JNK in a transcription-independent manner of p53

CHOE, YUN-JEONG,LEE, SUN-YOUNG,KO, KYUNG WON,SHIN, SEOK JOON,KIM, HO-SHIK Spandidos Publications 2014 International journal of oncology Vol.44 No.3

A recent study reported that p53 can induce HO-1 by directly binding to the putative p53 responsive element in the HO-1 promoter. In this study, we report that nutlin-3, a small molecule antagonist of HDM2, induces the transcription of HO-1 in a transcription-independent manner of p53. Nutlin-3 induced HO-1 expression at the level of transcription in human cancer cells such as U2OS and RKO cells. This induction of HO-1 did not occur in SAOS cells in which p53 was mutated and was prevented by knocking down the p53 protein using p53 siRNA transfection, but not by PFT-alpha, an inhibitor of the transcriptional activity of p53. Accompanying HO-1 expression, nutlin-3 stimulated the accumulation of ROS and the phosphorylation of MAPKs such as JNK, p38 MAPK and ERK1/2. Nutlin-3-induced HO-1 expression was suppressed by TEMPO, a ROS scavenger, and chemical inhibitors of JNK and p38 MAPK but not ERK1/2. In addition, nutlin-3-induced phosphorylation of JNK but not p38 MAPK was inhibited by TEMPO. Notably, the levels of nutlin-3-induced ROS were correlated with the mitochondrial translocation of p53 and this induction was prevented by PFT-beta, an inhibitor of the mitochondrial translocation of p53. Consistent with the effect of the ROS scavenger and MAPK inhibitors, PFT-beta reduced HO-1 expression and the phosphorylation of JNK induced by nutlin-3. In the experiments of analyzing cell death, the knockdown of HO-1 augmented nutlin-3-induced apoptosis. Collectively, these results suggest that nutlin-3 induces HO-1 expression via the activation of both JNK which is dependent on ROS generated by p53 translocated to the mitochondria and p38 MAPK which appears to be stimulated by a ROS-independent mechanism, and this HO-1 induction may inhibit nutlin-3-induced apoptosis, constituting a negative feedback loop of p53-induced apoptosis.

Dimethylnitrosamine 유발 급성 간 손상 흰쥐에서 ^(99m)Tc-Lactosylated Serum Albumin을 이용한 간 기능의 평가

정신영,이재태,서명랑,유정아,배진호,안병철,황재석,정재민,하정희,이규보 대한핵의학회 2003 핵의학 분자영상 Vol.37 No.6

목적: ^(99m)Tc-lactosylated serum albumin (^(99m)Tc-LSA)은 간세포에 특이적으로 결합하는 간수용체 영상용 방사성의약품으로 새로이 합성되었다. 간섬유화를 유발하는 dimethylnitrosamine (DMN)을 투여한 간 손상 휜쥐 모델에서 ^(99m)Tc-LSa의 역동학적인 간섭취를 조사하고 간효소치의 변화와 조직학적 소견을 비교하여, LSA의 간섭취가 간기능의 변화를 반영하는지를 연구하였다. 대상 및 방법: SD계 흰쥐에 DMN를 27 mg/kg으로 복강 내 주사하여 급성 간손상을 유도하고 대조군과 비교하였다. DMN을 주사한 흰쥐를 3일(DMN-3), 8일(DMN-8), 21일(DMN-21)에 ^(99m)Tc=LSA (1,665 mg/kg) 29 MBq를 정맥 주사하여, 30분 동안 동적 영상을 획득하고 간과 신장부위에 관심영역을 설정하여 간과 심장부위의 시간방사능 곡선을 얻었다. 간기능 평가를 위해 시간방사능 곡선을 이용하여 간섭취지수와 혈중제거지수를 구하였고 곡선 최적화를 시행하였다. DMN 투여군과 대조군의 간효소치의 변화와 간조직의 광학현미경 소견을 비교하였다. 결과: 대조군에서는 ^(99m)Tc-LSA가 빠르게 간에 섭취되고 혈중에서 제거되었으나 DMN을 처리한 군에서는 간섭취가 낮았다. 간섭취지수의 비교에서 대조군에 비해 DMN 처리군에서 유의하게 간섭취지수가 낮았다(DMN-3: 0.842, DMN-8: 0.898, DMN-21: 0.91, 대조군: 0.96, p<0.05). 혈중제거지수의 비교에서도 대조군에 비해 DMN 처리군에서 혈중제거지수가 유의하게 높았다(DMN-3: 0.731, DMN-8: 0.654, DMN-21: 0.604, 대조군: 0.473, p<0.05). 비선형 회귀분석에서 R_(2) 값은 0.9이상으로 좋은 일치를 보였고, 대조군에ㅓ K값이 DMN처리군에 비해 크고(DMN-3: 0.28, DMN-8: 0.41, DMN-21: 0.46, 대조군: 0.97, p<0.05), T_(1/2)값은 작았다(DMN-3: 2.5, DMN-8: 1.7, DMN-21: 1.5, 대조군: 0.7, p<0.05). 간효소치의 변화는 DMN-3군에서는 대조군에 비해 상승하였으나 DMN-8, DMN-21군에서는 간효소치의 상승이 관찰되지 않았다. 간조직 소견의 경우 DMN-3군에서 중심정맥 주위에 괴사가 관찰되었으나 DMN-8군, DMN-21군에서는 미약한 정도의 염증세포 침윤만이 관찰되었다. 결론: ^(99m)Tc-LSA 간신티그래피의 간섭취 정도는 간손상과 반비례하였으며 간섭취의 변화는 조직학적 손상이 심한 정도와 간손상후 회복되는 과정을 반영하여 주었다. ^(99m)Tc-LSA 간신티그래피가 간손상을 평가하고 간손상후 회복되는 과정을 추적하는 간수용체 영상용 방사성 의약품으로 사용될 수 있을 것으로 생각된다. Objects: ^(99m)Tc-lactosylated human serum albumin(LSA) is a newly synthesized radiopharmaceutical that binds to asialoglycoprotein receptors, which are specifically presented on the hepatocyte membrane. Hepatic uptake and blood clearance of LSA were evaluated in rat with acute hepatic injury induced by dimethylnitrosamine(DMN) and results were compared with corresponding findings of liver enzyme profile and these of histologic changes. Materials and Methods: DMN (27 mg/kg) was injected intraperitoneally in Sprague-Dawley rat to induce acute hepatic injury. At 3(DMN-3), 8(DMN-8), and 21(DMN-21) days after injection of DMN, LSA injected intravenously, and dynamic images of the liver and heart were recorded for 30 minutes. Time-activity curves of the heart and liver were generated from regions of interest drawn over liver and heart area. Degree of hepatic uptake and blood clearance of LSA were evaluated with visual interpretation and semiquantitative analysis using parameters (receptor index : LHL3 and index of blood clearance : HH3), analysis of time-activity curve was also performed with curve fitting using Prism program. Results: Visual assessment of LSA images revealed decreased hepatic uptake in DMN treated rat, compared to control group. In semiquantitative analysis, LHL3 was significantly lower in DMN treated rat group than control rat group (DMN-3:0.842, DMN-8: 0.898, DMN-21: 0.91, Control: 0.96, p<0.05), whereas HH3 was significantly higher than control rat group (DMN-3: 0.731, DMN-8: 0.654, DMN-21: 0.604, Control: 0.473, p<0.05). AST and ALT were significantly higher in DMN-3 group than those of control group. Centrilobular necrosis and infiltration of inflammatory cells were most prominent in DMN-3 group, and were decreased over time. Conclusion: The degree of hepatic uptake of LSA was inversely correlated with liver transaminase and degree of histologic liver injury in rat with acute hepatic injury.

Intracavitary Radiation Therapy for Recurrent Cystic Brain Tumors with Holmium-166-Chico : A Pilot Study

Ha, Eun Jin,Gwak, Ho-Shin,Rhee, Chang Hun,Youn, Sang Min,Choi, Chang-Woon,Cheon, Gi Jeong The Korean Neurosurgical Society 2013 Journal of Korean neurosurgical society Vol.54 No.3

Objective : Intracavitary injection of beta-emitting radiation source for control of cystic tumors has been tried with a benefit of localized internal radiation. The authors treated cystic brain tumor patients with Holmium-166-chitosan complex (Ho-166-chico), composed of a beta-emitting radionuclide Holmium-166 and biodegradable chit polymer, and evaluated the safety and effective measurement for response. Methods : Twenty-two patients with recurrent cystic brain tumor and/or located in a deep or eloquent area were enrolled in this pilot study. The cyst volume and wall thickness were determined on CT or MRI to assess radiological response. The activity of Ho-166-chico injected via Ommaya reservoir was prescribed to be 10-25 Gy to the cyst wall in a depth of 4 mm. Results : There was neither complications related to systemic absorption nor leakage of Ho-166-chico in all 22 patients. But, two cases of oculomotor paresis were observed in patients with recurrent craniopharyngioma. Radiological response was seen in 14 of 20 available follow-up images (70%). Seven patients of 'evident' radiological response experienced more than 25% decrease of both cyst volume and wall thickness. Another 7 patients with 'suggestive' response showed decrease of cyst volume without definitive change of the wall thickness or vice versa. All patients with benign tumors or low grade gliomas experienced symptomatic improvement. Conclusion : Ho-166-chico intracavitary radiation therapy for cystic tumor is a safe method of palliation without serious complications. The determination of both minimal effective dosage and time interval of repeated injection through phase 1 trial could improve the results in the future.

Antioxidant and Anti-Inflammatory Effects of Rhei Rhizoma and Coptidis Rhizoma Mixture on Reflux Esophagitis in Rats

Kwon, O Jun,Kim, Min Yeong,Shin, Sung Ho,Lee, Ah Reum,Lee, Joo Young,Seo, Bu-il,Shin, Mi-Rae,Choi, Hyun Gyu,Kim, Jeong Ah,Min, Byung Sun,Kim, Gyo-Nam,Noh, Jeong Sook,Rhee, Man Hee,Roh, Seong-Soo Hindawi Publishing Corporation 2016 Evidence-based Complementary and Alternative Medic Vol.2016 No.-

<P>The purpose of this study was to investigate the antioxidant and anti-inflammatory effects of the combined extract of Rhei rhizoma and Coptidis rhizoma (RC-mix) in experimental model of acute reflux esophagitis. The antioxidant activity was assessed by<I> in vitro</I> 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. RC-mix was given at 100, 200, and 400 mg/kg body weight 2 h prior to induction of reflux esophagitis (RE). After 5 h, the effects of RC-mix treated rats were compared with those of normal and control rats. The representative flavonoid contents of RC-mix, such as sennoside A, epiberberine, coptisine, palmatine, and berberine, were detected using HPLC. The elevated esophageal mucosa damage was markedly ameliorated by RC-mix treatment in a dose-dependent manner. Furthermore, the administration of RC-mix reduced the increase of serum reactive oxygen species (ROS) and peroxynitrite (ONOO<SUP>−</SUP>). The improvement of superoxide dismutase (SOD) and heme oxygenase-1 (HO-1) levels were marked in the group given RC-mix. Moreover, the elevation of inflammatory mediators and cytokines by nuclear factor-kappa B (NF-<I>κ</I>B) activation in control rats decreased by RC-mix pretreatment. These results indicate that RC-mix treatment reduces the pathological states of esophagitis<I> via</I> regulating NF-<I>κ</I>B mediated inflammation related to oxidative stress. </P>

Nrf2-Heme oxygenase-1 modulates autophagy and inhibits apoptosis triggered by elevated glucose levels in renal tubule cells

Joo Heon Kim,김경민,Jin Uk Jeong,Jong-Ho Shin,신재민,방기태 대한신장학회 2019 Kidney Research and Clinical Practice Vol.38 No.3

Background: Autophagy is a highly balanced process in which lysosomes remove aged and damaged organelles and cellular proteins. Autophagy is essential to maintain homeostasis in the kidneys. Methods: Using human renal tubule cells HK-2, we assessed the impact of high glucose (HG) on autophagy. We also evaluated the capability of sulforaphane (SFN) to protect the HK-2 cells from HG-induced apoptosis by modulating autophagy. Results: SFN modulated autophagy and decreased apoptosis in the HK-2 cells that were cultured in 250 mM glucose medium for two days. The reactive oxygen species (ROS) levels increased, as expected, in the cells cultured in the 250 mM glucose medium. However, the SFN decreased the ROS levels in the HK-2 cells. The overexpression of heme oxygenase-1 (HO-1) by SFN decreased the expression of LC3 and beclin-1. LC3 and beclin-1 were involved in the downregulation of caspase-3 that was observed in the HG-induced cells. Conclusion: The activation of nuclear factor E2-related factor 2 (Nrf2)-HO-1 inhibited ROS expression and subsequently attenuated autophagy and cell apoptosis after HG injury was decreased. HG injury led to the activation of autophagy and HO-1 in order to combat oxidative stress and protect against cell apoptosis. Therefore, HO-1 activation can prevent ROS development and oxidative stress during HG injury, which considerably decreases autophagy and apoptosis.

특발성 경련질환으로 오인된 원발성 부갑상선기능저하증 1례

신성하,최정영,김준기,고영춘,조상욱,김순호,임중규 全南大醫大 2001 전남의대학술지 Vol.37 No.1

머리부분에 방사선조사를 받은 흰쥐 샘뇌하수체의 변화에 대한 면역전자현미경적 연구

신기호,박경호,안의태,양남길,고정식 순천향의학연구소 1995 Journal of Soonchunhyang Medical Science Vol.1 No.2

본 실험은 과다한 X-선에 머리부분이 노출되었을 때, 샘뇌하수체의 변화를 알아보기 위하여 시행하였다. 체중 200-250g의 Sprague Dawley계 숫흰쥐를 실험동물로 사용하였으며, 정상군과 방사선조사군으로 나누었다, 방사선조사군은 조사량에 따라 3,000 rad 조사군과 6,000 rad조사군으로 나누어, 방사선 조사후 6시간, 2일 및 6일 후에 도살하여 조직을 절취하였다. 방사선조사는 흰쥐를 sodium thiopental로 마취한 후 방사선선형가속기(Mitsubishi Linear Accelerator ML-4MV)를 사용하여 머리부위를 조사하였다. 조사조건은 조사거리 80 cm, 조사구역 30 cm X 30 cm, 조사깊이 1.2 cm(100% skin dose)였으며, 분당 200 rad씩 조사하였다. 샘뇌하수체는 1% glutaraldehyde- 1% paraformaldehyde액으로 일차 고정한후, 2% osmium tetroxide액에 이차고정하였으며, 고정이 끝난 조직은 alcohol과 acetone으로 탈수한 후 araldite혼합액에 포매하였다. 포매된 조직은 , LKB-V ultratome으로 60-70nm두께의 절편을 작성하여 300 mesh nickel grid에 붙인 다음 젖샘자극호르몬과 성장자극호르몬에 대한 단독면역염색 및 이중면역염색을 시행하였다. 면역염색이 끝난 절편은 uranyl acetate와 lead citrate로 염색한후, JEM 100CX-Ⅱ 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 1. 젖샘자극호르몬분비세포는 불규칙한 모양을 한 큰 분비과립(300-700 nm)을 가진 성숙세포, 크기가 다양한 둥근 분비과립(150-200 nm)을 가진 중간 세포와 크기가 작은 둥근 분비과립(100-150 nm)을 가진 미성숙세포로 나눌 수 있었다. 2. 성장자극호르몬분비세포는 크고 둥근 분비과립(200-500 nm)을 가진 제 1 형 세포와 상대적으로 작고 둥근 분비과립 (150-200 nm)을 가진 제 Ⅱ형 세포로 나눌 수 있었다. 3. 방사선 조사후 6 시간군에서 3,000 rad에서는 큰 변화가 없었고, 6,000 rad에서는 사립체와 과립형질내세망의 수조 확장이 관찰되었다. 방사선 조사후 2일군에서는 큰 변화가 없었으며, 6일군에서는 핵막구조의 확장이 관찰되었다. 4. 세포의 종류도 젖샘자극호르몬분비세포는 3,000 rad 조사군과 6,000 rad 조사군 모두 2일군에서는 성숙형이 감소하고 중간형과 미성숙형이 자주 관찰되었으며, 6일군에서는 정상군과 같은 분포양상을 보였으나 분비과립의 금입자표지가 감소한 것을 관찰할 수 있었다. 성장자극호르몬분비세포에서는 뚜렷한 세포형이 분포변화는 관찰할 수 없었으나 금입자표지의 감소는 나타났다. 5. 방사선조사후 6시간군에서 부터 mammosomatography가 나타났는데, 한 세포 내에 젖샘자극호르몬과 성장자극호르몬을 지닌 분비과립이 함께 존재하였으며, 세포의 모양은 다핵세포의 형태를 하고 있는 것과 불규칙한 모양을 한 것이 있었는데, 이와같은 결과는 방사선조사 후 샘뇌하수체의 기능저하에 따른 보상작용으로 나타난 현상이라 생각된다. 이상의 결과로 보아 방사선 조사를 받은 초기에는 분비과립의 방출이 과다하게 일어나며, 방사선 조사의 영향으로 세포질소기관의 기능이 약화되어 6일이 지나면 샘뇌하수체의 호르몬분비능력이 저하되는 것 같다. This experiment was performed to study the morphological changes of the adenohypophysis of rat following X-ray irradiation. Male rats were divided into normal and X-ray irradiation groups. The heads of rat were exposed to 3,000 rads or 6,000 rads of radiation in a single dose. X-ray source was a Mitsubishi Linear Accelerator ML-4MV. Only the heads of animals were exposed at the distance of 80 cm, within the area of 30 X 30 cm, in the depth of 1 cm, with the speed of 200 rad/min. Animals of X-ray irradiation group were sacrificed on 6 hours, 2 days and 6 days after the irradiation. Tissue blocks of adenohypophysis were fixed in the 1% glutaraldehyde - 1% paraformaldehyde solution, followed by refixation in the 2% osmium tetroxide solution. Dehydradted blocks were embedded in araldite mixture. The sections were cut on a LKB V ultrotome, and ultrathin sections were places on bare nickel grid(200 mesh). The section-bearing grids were floated upside down on the solutions in a moisture chamber at room temperature. Sections were single immunostained or double immunostained for prolactin and/or growth hormone. And the sections were jet washed with distilled water. The immunostained sections were contrasted with uranyl acetate and lead citrate, and observed with JEM 100CX II electron microscope. The results were as follows: 1. Three types of the prolactin cells according to their size and shape of secretory granules were found; mature type cells contained large pleomorphic secretory granules(above 500 nm). intermediate type cells contained round granules of varying size(200-250 nm), and immature type cells contained small round granules(100 nm). 2. Two types of the growth hormone cells according to their size of secretory granules were found: type I cells contained large round secretory granules(200-300 nm) and type II cells contained small granules(100-200 nm). 3. Six hours after the irradiation, adenohypophysis showed swollen cisternae of granular endoplasmic reticulum and alterations of mitochondrial cristae, but only in doses of 6.000 rads. 4. Adenohypopysis showed decrease in number of mature type prolactin cells on 2 days after irradiatin, and recovered number of mature typer prolactin cell. but their immunoreactiveity were gradually decreased following the irradiation, as compared with that in normal adenohypophysis. 5. Six hours after the irradiation, mammosomatotrophs were found to contain prolactin and growth hormone within different granules each other. Some cells were multinucleated, and some cells exhibited irregular cytoplasmic processes. Summarizing the above results, adenohypophysis rapidly released the secretory granules after the irradiation, and cell organelles of prolactin cells and growth hormone cells were altered or degenerated. It means that adenohypophysis decreased its hormone producing activity on 6 days after irradiation.

콩나물의 asparagine 함량에 미치는 요인 구명

정연신,손태호,Krishna Hari Dhakal,이정동,황영현 경북대학교 농업과학기술연구소 2008 慶北大農學誌 Vol.26 No.-

알코올의 숙취해독에 뛰어난 효과가 있는 것으로 알려진 sparagine은 콩나물의 뿌리에 함량이 가장 많은 것으로 알려져 있다. 본 연구논문에서는 콩나물 뿌리 발생에 미치는 재배실의 온도, 일당 관수횟수, 재배일수 및 동일용기에 사용한 콩 량에 대한 영향을 조사하여 얻어진 연구결과를 요약하면 다음과 같다. 1. 아가통과 풍산나물콩의 수율은 재배일수에 따라 경시적으로 증가하였으며, 재배실 온도가 20℃에서 보다 30℃에서 높았으며, 콩나물의 수율은 5회 관수, 8일 재배시 가장 높았다. 2. 아가콩과 풍산나물콩의 콩나물 부위별 건물중의 비율은 자엽이 가장 많았고 뿌리가 가장 낮았으며, 재배실 온도가 20℃에서 보다 30℃에서 비율이 더 높았다. 3. 아가콩의 sparagine 함량은 재배실 온도 30℃에서 5회 관수 8일 재배한 하배축에서 18.9%로 가장 높게 나타났다. 4. 풍산나물콩의 sparagine 함량은 재배일수가 길수록, 재배온도가 높은 경우 높았으며, 하배축에서 가장 높았고, 30℃에서 2회 관수 8일 재배시 하배축에서 18.8%로 가장 높게 나타났다. 5. 아가콩은 300g, 풍산나물콩은 500g 치상시 수율이 가장 높았으며, 잔뿌리의 수는 아가콩과 풍산나물콩 모두 치상량이 증가함에 따라 증가하는 경향이었다. It is known that the content of asparagine, an excellent detoxifying substance of alcohols in human body, is the highest in the roots of soy-sprout. At the same time, it is inferred that soy-sprouts producing more roots are better for detoxifying. In this experiment, the effects of room temperatures on number of watering per day, and duration of soy-sprout culture were carefully investigated. Some of the results obtained are as follows; 1. The yield rate of soy-sprouts for Agakong and Pungsannamulkong was continuously increased up to 9 days. It was higher in room temperature of 30℃ than in 20℃ and was the highest at 8th day of culture with 5 times of watering per day. 2. The asparagine content in soy-sprouts of Agakong and Pungsannamulkong was the highest in cotyledon and the lowest in roots. This rate was higher in the room temperature of 20℃ than in 30℃. 3. The highest asparagine content of soy-sprout of Agakong was 18.9%, obtained in the room temperature of 30℃, cultivated for 8 days with 5 times of watering per day. 4. The highest asparagine content of Pungsannamulkong was 18.8%, obtained in hypocotyl cultivated in the room temperature of 30℃ for 8 days with the number of 2 times watering per day. 5. When an cultivation apparatus of 5 liters volume was used, the optimum seed amount for the highest yield rate was 300g for Agakong and 500g for Pungsannamulkong. At the same time, the number of lateral roots showed increasing tendency with more amounts of soybean seeds used.

R.F. Magnetron Sputtering법으로 제조한 LaCoO₃박막의 가스감지 특성

신정호,김정규,박기철,전춘배,장재영 慶尙大學校 工科大學 自動化및컴퓨터應用技術硏究所 1998 自動化 및 컴퓨터應用技術 Vol.5 No.1

LaCoO₃(LCO) thin film sensors were fabricated on an alumina substrate by r.f. magnetron sputtering. By varying the annealing temperature of the LCO film from 600℃ to 1000℃, we investigated X-ray diffraction pattern, electrical properties, and gas sensing properties of the films. The crystallinity was improved and the activation energy was increased with increasing the annealing temperature. The sensitivities of the LCO films for NH₃ and CO gases were also increased with the annealing temperature. Sensing properties of LCO thin films were improved when the measuring temperature was 350℃ or more

직무중심의 성과주의 인사제도

신건호,백정식 단국대학교 경영경제연구소 2000 經營, 經濟硏究 Vol.2 No.-

Corporate's mission is to creat values in market, to develop and to prolong. Management is to evaluate, to actualize values effectively in market. The thinking of outcomes in corporate has been changed from sales amount, profits to social contribution, environmental creation. The key of innovation is performance - The idea of intellectual performance. The management based performance focuses on the effective and high quality performance in individuals of organizations. The future role of H.R.M. is to develop High Performer at last, to raise the corporate's power of value creation. The realization of corporate's mission comes true through employee's job. Accountability is composed of performance and activity. the personnel management based job classification is quite different from person related. The most important definitions are Accountability and Competency. Job based compensation is different from Seniority based compensation. Job based compensation will actualize the corporate's needs for exemple reduction of persennel expenses and motivation of young employees. Performance Management System has an important role in Pay for Performance system. Competency is another important ability and characteristics to create performance. To recruit and to transfer someone who have motivation and characteristics, to teach them skills and knowledge will be realistic and effective.