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.

Schisandrin C enhances odontoblastic differentiation through autophagy and mitochondrial biogenesis in human dental pulp cells

Takanche, Jyoti Shrestha,Kim, Jeong-Seok,Kim, Ji-Eun,Han, S-H.,Yi, Ho-Keun Elsevier 2018 Archives of oral biology Vol.88 No.-

<P><B>Abstract</B></P> <P><B>Objective</B></P> <P>To investigate the role of Schisandrin C in odontoblastic differentiation, and its relations between autophagy and mitochondrial biogenesis in human dental pulp cells (HPDCs).</P> <P><B>Design</B></P> <P>Fresh third molars were used, and cultured for HDPCs. Western blotting technique, Alizarin red S staining, alkaline phosphatase (ALP) activity, and confocal microscopy were used to detect autophagy, mitochondrial biogenesis, and odontoblastic differentiation. To understand the mechanism of Schisandrin C, the HDPCs were treated with lipopolysaccharide (LPS), autophagy and heme oxygenase-1 (HO-1) inhibitors: 3-Methyladenine (3-MA) and Zinc protoporphyrin IX (ZnPP), respectively.</P> <P><B>Results</B></P> <P>LPS decreased the expression of autophagy molecules [autophagy protein 5 (ATG-5), beclin-1, and microtubule-associated protein 1A/1B light chain 3 (LC3-I/II)] and mitochondrial biogenesis molecules [heme oxygenase-1 (HO-1) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α)], and disrupted odontoblastic differentiation. The down-regulation of autophagy and mitochondrial biogenesis with 3-MA and ZnPP inhibited odontoblastic differentiation. However, Schisandrin C restored the expression of all the above molecules, even with LPS and inhibitor treatment. This result demonstrates that autophagy and mitochondrial biogenesis plays an essential role in odontoblastic differentiation, and Schisandrin C activates these systems to promote odontoblastic differentiation of HDPCs.</P> <P><B>Conclusion</B></P> <P>Schisandrin C has potential characters to regulate odontoblastic differentiation, and may be recommended for use as a compound for pulp homeostasis.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Autophagy and mitochondrial biogenesis are linked with odontoblastic differentiation. </LI> <LI> Schisandrin C promotes odontoblastic differentiation in HDPCs. </LI> <LI> It mediated this function via mitochondrial biogenesis and autophagy. </LI> </UL> </P>

水素 이온 濃度變化가 平滑筋 細胞膜에서의 KCI 誘導 Ca-Uptake에 미치는 影響

昔廷鎬,林鍾鎬,李載欣 충남대학교 의과대학 지역사회의학연구소 1986 충남의대잡지 Vol.13 No.1

For the purpose of studying the properties of calcium channel, the effect of pH on the KCl induced Ca-uptake in the isolated porcine intestinal sarcolemmal vesicles, which were loaded with high KCl was investigated. The obtained results were as follows; 1. Ca-uptake in the membrane of depolarized state by high KCl, compared with that of polarized state by low KCl in the medium was increased significantly(increased about 95.7% for 30 seconds, pH 7.4). 2. Ca-uptake in the membrane of predepolarized state by high KCl, compared with that of polarized state by low KCl was also increased significantly but less than that of depolarized state by high KCl (increased about 66.7% for 30 seconds, pH 7.4). 3. Influence of pH on the Ca-uptake of various membrane state was increased or decreased according to pH changes. Ca-uptake in the membranes of incubated at pH 6.0 was decreased about 48.5%, but increased about 10% and 36% at pH 8.0 and 9.0 respectively, compared with that incubated at pH 7.4. 4. KCl-induced Ca-uptake at the various pH conditions, was not inhibited by the pretreatment of diltiazem(10^-6M). From the above results, it was concluded that KCl-induced Ca-uptake at the various pH conditions in the isolated sarcolemmal membranes, prepared from porcine intestinal smooth muscle, was not mediated through channels which are inhibited by organic Ca-antagonists.

고혈압쥐 적혈구 및 혈관조직에서의 Rubidium 이동과 Ouabain 결합에 관한연구

석정호,허강민,이재흔 충남대학교 의과대학 지역사회의학연구소 1989 충남의대잡지 Vol.16 No.1

To study the alterations of Na-pump in the 1-kidney, 1-clip hypertensive rat, made by partial ligation of left renal artery and by removal of right kidney, we determined Na, K-ATPase activity in the RBC and carried out the experiment of Rb-uptake and H-ouabain-binding from the RBC and aorta. Results were as follows; 1. Blood pressure was significantly increased to 117.4/121.6mmHg in 1-kidney, 1-clip hypertensive rat as 4 weeks after operation. 2. Na, K-ATPase activity in the RBC membrane of hypertensive rat was significantly decreased, compared to that of control rat(P<0.05). 3. Ouabain-sensitive Rb-uptake was significantly decreased in the RBC and aorta strip on hypertensive rat, compared to that of the control rat. 4. Bmax of the RBC and aorta strip in the ouabain-binding study was significantly decreased, but values of Kd was significantly increased in hypertensive rat. From these results, it is suggested that decreased activity of Na-pump in hypertensive rat RBC and aorta strip, is due to the alteration of number and property of Na-pump.

Chloroform 만성 중독으로 인한 간병변에 대한 병리학적 연구

정호석,강대영 忠南大學校 癌共同硏究所 1991 癌共同硏究所 硏究誌 Vol.1 No.1

In an attempt to elucidate the pathological effects of chloroform(CHCl_(3)) administration, the present study was undertaken in male Sprague-Dawley rats. Evidence for the existence of hyperplastic nodules, oval cells, bile duct or ductular proliferation, and liver cirrhosis was examined, and the number of dysplastic cells were evaluated, emphasizing especially on the difference among the several groups. Rats were administered 300mg/kg or 600mg/kg of chloroform(CHCl_(3)-corn oil 1:4 solution) intraperitoneally, and then twice injected weekly for 36 weeks. The control animals were given as similar volume of saline or corn oil, respectively. The animals were sacrificed in 3rd, 6th and 9th month after chloroform injection. The histopathologic changes in the liver of the control and experimenal groups were noted as follows: 1) The hyperplastic nodules in the 600 mg/kg chloroform treated group were more prominent than those of the 300 mg/kg chloroform treated group in the number and size. 2) The number of dysplastic hepatocytes were increased gradually as the experiment continiued. The incidence of dysplastic hepatocytes was 45.5% in the 300 mg/kg treated group and 70.6% in the 600 mg/kg treated group. 3) Oval cells appeared in the early days of the experiment (3rd month) and then gradually decreased in number. 4) The degree of severity of liver cirrhosis and bile duct or ductular proliferation gradually increased as the experiment continued. Incidence of liver cirrhosis and bile duct or ductular proliferation was 31.8% in the 300 mg/kg treated group and 64.7% in the 600 mg/kg treated group, respectivley. 5) Incidence of the chloroform induced hepatoma was 5.9%(1 out of 17 rats) in the chloroform 600 mg/kg treated group, and the hepatoma spread to the liver, followed by metastasis to the omentum and ileum. 6) Light microscopically, some hepatic lobules revealed some foci of vacuolated or clear hepatocytes. Electron microscopically, they contained fat droplets. Others also contained increased number of mitochondria or distorted mitochondria. In summary, the result obtained by the present study indicates chloroform induced hyperplastic nodules, liver cell dysplasia, oval cell proliferation and liver cirrhosis, and secondarily induced hepatoma.

側腦室內 Substance P가 家兎 腎臟機能에 미치는 影響

昔延鎬,黃炳斗,李載欣 충남대학교 의과대학 지역사회의학연구소 1981 충남의대잡지 Vol.8 No.2

In this study, the effects of substance P, administered intracranioventricularly, on the renal function of the rabbit were investigated. 1) Intraventricular substance P(0.01, 0.1 and 10ng) elicited antidiuresis, and decreased the renal plasma flow, glomerular filtration rate and urinary excreted amounts of sodium and potassium, but did not influence to free water clearance. 2) Correlation coefficient between changes of urine volume and of renal plasma flow, glomerular filtration rate or urinary sodium excretion was statistically significant. 3) Systemic blood pressure was not affected in a dose of 0.01ng, but increased in 0.1ng(10mmHg) or 10ng(20∼25mmHg). 4) Antidiuretic effects of intraventricular substance P were partially inhibited by the renal nerve degeneration. From the above results, it is suggested that the antidiuretic effect of intraventricular substance P is dae to renal hemodynamic changes, partially induced by renal sympathetic nerve activation.

안와 내벽 외향 골절의 내시경적 비내 재건술

홍석찬,진정언,강성호 건국대학교 의과학연구소 1997 건국의과학학술지 Vol.- No.7

자발성 뇌출혈 환자의 일상생활동작 수행에 영향을 미치는 인자

정호중,진기은,박희석 高神大學 醫學部 1996 高神大學醫學部 論文集 Vol.11 No.1-2

側腦室內 r-Aminobutyric Acid가 家兎賢臟機能에 미치는 影響

昔廷鎬 충남대학교 의과대학 지역사회의학연구소 1979 충남의대잡지 Vol.6 No.2

As a result of investigation to various changes of renal function after GABA administered into lateral ventricle of the rabbit, following results were obtained. 1. Urine flow, glomerular filtration rate, renal blood flow and urinary excretion of sodium and potassium were decreased in 20 minutes after intraventricular GABA(30 or 100㎍). And correlation coefficients between two parameter changes among them were statistically significant, respectively. 2. Intravenous GABA (100 ㎍) was not effect to renal function. 3. 30㎍ intraventricular GABA was not influenced to blood pressure, while 100㎍ GABA was transient decrease in blood pressure(15mmHg) but recovered within 10 minutes. 4. The effect of intraventricular GABA on the renal function was inhibited after regitine pretreatment (2mg/kg) intravenously. From the above results, it is suggested that the effect of intraventricular GABA on the renal function of rabbit is due to renal hemodynamic changes by central sympathetic stimulation.