N-acetyl cysteine inhibits H2O2-mediated reduction in the mineralization of MC3T3-E1 cells by down-regulating Nrf2/HO-1 pathway

( Daewoo Lee ),( Sung Ho Kook ),( Hyeok Ji ),( Seung Ah Lee ),( Ki Choon Choi ),( Kyung Yeol Lee ),( Jeong Chae Lee ) 생화학분자생물학회(구 한국생화학분자생물학회) 2015 BMB Reports Vol.48 No.11

There are controversial findings regarding the roles of nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway on bone metabolism under oxidative stress. We investigated how Nrf2/HO-1 pathway affects osteoblast differentiation of MC3T3-E1 cells in response to hydrogen peroxide (H2O2), N-acetyl cysteine (NAC), or both. Exposing the cells to H2O2 decreased the alkaline phosphatase activity, calcium accumulation, and expression of osteoblast markers, such as osteocalcin and runt-related transcription factor-2. In contrast, H2O2 treatment increased the expression of Nrf2 and HO-1 in the cells. Treatment with hemin, a chemical HO-1 inducer, mimicked the inhibitory effect of H2O2 on osteoblast differentiation by increasing the HO-1 expression and decreasing the osteogenic marker genes. Pretreatment with NAC restored all changes induced by H2O2 to near normal levels in the cells. Collectively, our findings suggest that H2O2-mediated activation of Nrf2/HO-1 pathway negatively regulates the osteoblast differentiation, which is inhibited by NAC. [BMB Reports 2015; 48(11): 636-641]

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.

Prostaglandin A₂-induced Apoptosis is Not Inhibited by Heme Oygenase-1 in U2OS Cells

Kyoung-Won Ko(고경원),Sun-Young Lee(이선영),Ji-Hyun Ahn(안지현),Jaetaek Kim(김재택),In-Kyung Kim(김인경),Ho-Shik Kim(김호식) 한국생명과학회 2008 생명과학회지 Vol.18 No.11

Prostaglandin A₂ (PGA₂)는 사람 골육종 세포인 U2OS 세포주에서 apoptosis와 heme oxygenase (HO)-1의 발현을 함께 유도하였다. PGA₂에 의한 apoptosis는 HO-1의 과도한 발현이나 HO-1에 대한 small interfering RNA에 의한 발현저하에 의하여 변동되지 않았으나 H₂O₂에 의한 세포사망은 HO-1의 발현 수준에 반비례하여 변동되었다. 또한 thiol antioxidant인 N-acetyl-L-cysteine (NAC)은 PGA₂에 의한 세포사망과 HO-1의 발현 증가를 모두 차단하였지만, non-thiol antioxidant인 butylated hydroxyanisole (BHA)과 ascorbic acid는 세포사망과 HO-1의 발현 유도를 차단하지 않았다. 이와 같은 결과들은 PGA₂는 산화성 손상에 의해서가 아니라 PGA₂의 thiol-reactivity에 의하여 apoptosis와 HO-1의 발현을 유도하며, HO-1의 발현은 PGA₂에 의한 apoptosis와는 독립적인 현상이거나 기능적으로 apoptosis 유도의 하부에 위치하고 apoptosis의 진행에는 기여하지 않을 것이라는 것을 시사해 준다. Prostaglandin A₂ (PGA₂), one of cyclopentenone PGs, induced both apoptosis and heme oxygenase (HO)-1 expression in U2OS cells. PGA₂-induced apoptosis was not perturbed by either over-expression or knock-down of HO-1, whereas H₂O₂-induced cell death was inversely modulated by the expression level of HO-1. In addition, N-acetyl-L-cysteine (NAC), a thiol antioxidant, blocked both apoptosis and HO-1 expression induced by PGA₂. But, non-thiol antioxidants like butylated hydorxyanisole (BHA) and ascorbic acid did not block either apoptosis or HO-1-induction. Taken together, these results suggest that PGA₂ induces both apoptosis and HO-1 expression, which are critically related to the thiol-reactivity of PGA₂, but not oxidative stress, and HO-1 expression may be independent or functionally located downstream of apoptosis by PGA₂ without contribution to apoptosis progression.

기종성 신우신염 1례

이재욱,구정태,서정일,양창헌,이정호,이동철,이영현,이활,이경섭 동국대학교 경주대학 1997 東國論集 Vol.16 No.1

기종성 신우신염은 주로 당뇨병환자나 폐쇄성 요로질환을 가진 환자에 발생하며 특징적으로 신실질 및 그 주위조직에 가스를 형성하면서 심한 조직괴사를 일으키는 매우 드문 급성 화농성 신감염으로 사망률이 높은 질환이다. 이 질환은 1989년 Kelly와 MacCallum에 의해 처음 보고된 이래 외국문헌에 약 90례 정도가 보고되어 있으며 국내에는 28례가 보고되어 있다. 수액 및 전해질 보충, 인슐린 투여를 통한 적절한 혈당조절 및 항생제 투여등의 내과적 치료와 절개배농 및 신적출술등의 수술적 치료 방법이 있다. 내과적 치료로 가스의 감소가 없는 경우에는 즉각적인 수술적 치료를 시행하여야하므로 이 가스변화에 대한 추적관찰이 중요하다 하겠다. 본 저자들은 당뇨병환자에서 발생한 기종성 신우신염 1례를 내과적 요법을 치험하였기에 문헌고찰과 함께 보고하고자 한다. Emphysematous pyelonephritis is rare, life-threatening infection of the renal parenchyma and perirenal tisseue. This disease is characterized by the production of intrarenal and perirenal gas and is frequently encountered in patients with diabetes mellitus or urinary obstruction. We experienced a case of emphysematouse pyelonephritis in a 62 years old women with poorly controlled diabetes mellitus who had been managed with medical theraphy. So we report this case with a review of the referenced literatures.

췌장 선방세포암 1예

이화정,지준호,박승찬,박정철,최은정,서혜진,이원식,이정림,배병조,손경락,이경희 영남대학교 의과대학 2008 Yeungnam University Journal of Medicine Vol.25 No.2

Acinar cell carcinoma is a rare tumor that represents 1~2% of al1 pancreatic cancers. Clinical and radiologic findings are inconclusive in this disease Acinar cell carcinoma is characterized by rapid progression and early metastasis, which lead to its poor prognosis. A 41-year-o1d man was admitted to our hospital for abdominal pain. Abdominal computed tomography (CT) and positron emission tomography-computed tomography (PET-CT) showed a splenic mass, which was being invaded by a pancreatic tail mass and which had increased ^(18)F-fluorodeoxyglucose (FDG) uptake Primary radical distal pancreatectomy and splenectomy were performed. Pathologic findings revealed an acinar cell carcinoma of the Pancreas The patient underwent a total gastrectomy three months later because of gastric recurrence Four months later, multiple hepatic metastases were discovered, and the patient underwent a left hepatectomy During treatment with capecitabine, there was no evidence of tumor progression for 14 months. We report a case of metastatic pancreatic acinar cell carcinoma, which did not progress for an extended period while the patient was being treated with capecitabme.

Hepatoprotective and Antioxidative Activities of <i>Cornus officinalis</i> against Acetaminophen-Induced Hepatotoxicity in Mice

Lee, Nam-Hun,Seo, Chang-Seob,Lee, Ho-young,Jung, Da-Young,Lee, Jun-Kyung,Lee, Jin-Ah,Song, Kye Yong,Shin, Hyeun-kyoo,Lee, Mee-Young,Seo, Young Bae,Kim, Hokyoung,Ha, Hyekyung Hindawi Publishing Corporation 2012 Evidence-based Complementary and Alternative Medic Vol.2012 No.-

<P>The fruit of <I>Cornus officinalis </I>Sieb. et Zucc. is commonly prescribed in Asian countries as a tonic formula. In this study, the hepatoprotective effect of ethanolic extracts of the fruit of <I>C. officinalis</I> (ECO) was investigated in a mouse model of acetaminophen- (APAP-) induced liver injury. Pretreatment of mice with ECO (100, 250, and 500 mg/kg for 7 days) significantly prevented the APAP (200 mg/kg) induced hepatic damage as indicated by the serum marker enzymes (AST, ALT, and LDH). Parallel to these changes, ECO treatment also prevented APAP-induced oxidative stress in the mice liver by inhibiting lipid peroxidation (MDA) and restoring the levels of antioxidant enzymes (SOD, CAT, and HO-1) and glutathione. Liver injury and collagen accumulation were assessed using histological studies by hematoxylin and eosin staining. Our results indicate that ECO can prevent hepatic injuries associated with APAP-induced hepatotoxicity by preventing or alleviating oxidative stress.</P>

정신분열병에 대한 리스페리돈의 효과 및 안정성

이민수,김용구,김영훈,연병길,오병훈,윤도준,윤진상,이철,정희연,강병조,김광수,김동언,김명정,김상훈,김희철,나철,노승호,민경준,박기창,박두병,백기청,백인호,손봉기,손진욱,양병환,양창국,우행원,이정호,이종범,이홍식,임기영,전태연,정영조,정영철,정인과,정인원,지익성,채정호,한상익,한선호,한진희,서광윤 大韓神經精神醫學會 1998 신경정신의학 Vol.37 No.1

연구목적 : 본 시험의 목적은 임상시험 시작전에 연구자들을 대상으로 PANSS Workshop을 통하여 PANSS, ESRS에 대한 국내에서의 표준화 작업을 구축하고 새로운 정신병 치료제인 리스페리돈의 효과와 안정성을 재확인하여 리스페리돈 사용에 대한 적정화를 이루는데 있다. 연구방법 : 1996년 4월부터 1996년 9월까지 국내 39개 대학병원 정신과에 입원중인 혹은 증상이 악화되어 입원하는 정신분열병 환자 377명을 대상으로 다시설 개방 연구를 시행하였다. 1주일간의 약물 배설기간을 가진후, 리스페리돈을 8주간 투여하였고, 기준점, 1주, 2주, 4주, 그리고 8주후에 평가되었다. 용량은 제1일에는 리스페리돈 1mg씩 1일 2회, 제2일에는 2mg씩 1일 2회, 제3∼7일에는 3mg씩 1일 2회 투여하였다. 이후 환자의 임상상태에 따라 임의로 증량할 수 있으며, 최대 일일 16mg을 초과하지 않도록 하였다. 추체외로 증상을 조절하기 위한 투약을 허용하였다. 임상증상 및 부작용의 평가는 PANSS(Positive and Negative Syndrome Scale), CGI(Clinical Global Impression) 그리고 ESRS(Extrapyramidal Symptom Rating Scale)을 사용하였다. 연구결과 : 377명중 343명(91%)이 8주간의 연구를 완결하였다. 치료 종결시점인 8주후 PANSS 총점수가 20% 이상 호전된 경우를 약물 반응군으로 정의할때, 약물반응군은 81.3%였다. 리스페리돈에 반응하는 예측인자로는 발병연령, 이전의 입원 횟수, 유병기간이 관련 있었다. 리스페리돈은 1주후부터 PANSS양성, 음성, 및 일반정신병리 점수상에 유의한 호전을 보여 효과가 빨랐다. CGI의 경우도 기준점에 비해 1주후부터 유의한 감소를 나타내었다. ESRS의 경우, 파킨슨 평가점수는 기준점과 비교해 투여 1주, 2주, 4주후 유의하게 증가되었다가 8주후 기준점과 차이가 없었다. Dystonia 평가점수는 1주후만 유의한 증가를 보였으며, dyskinesia 평가점수는 유의한 차이가 없었다. 혈압, 맥박수의 생명징후 및 일반 혈액학 검사, 생화학적 검사, 심전도 검사에서 유의한 변화는 없었다. 결 론 : 이상의 다시설 개방 임상 연구를 통해 리스페리돈은 정신분열병 환자에서 양성증상뿐만 아니라 음성증상 및 전반적인 증상에도 효과적인 것으로 사료된다. 보다 명확한 평가를 위해서는 다른 항정신병약물과의 이중맹검 연구가 필요할 것으로 생각되며, 또한 장기적 치료에 대한 평가도 함께 이루어져야 하겠다. Objective : The purpose of this study was to investigate the efficacy and safety of risperidone in the treatment of Korean schizophrenic patients. Method : This multicenter open study included 377 schizophrenic patients drawn from 39 university hospitals. After a wash-out period of 1 week, the schizophrenic patients were treated with risperidone for 8 weeks and evaluated at 5 points ; at baseline, and 1, 2, 4 and 8 weeks of treatment. The dose was increased from 2mg/day(1mg twice daily) to 6mg/day(3mg twice daily) during the first week and adjusted to a maximum of 16mg/day over the next 7 weeks according to the patient's clinical response. Medication to control extrapyramidal symptoms was permitted. The psychiatric and neurological status of the patients was assessed by PANSS, CGI, and ESRS scales. Results : 343(91%) of 377 patients completed the 8-week trial period. Clinical improvement, as defined by a 20% or more reduction in total PANSS score at end point, was shown by 81.3% of patients. The predictors of response to risperidone were associated older age, shorter duration of illness, fewer previous hospitalization. Risperidone had rapid onset of action ; a significant decrease of the total PANSS and three PANSS factor(positive, negative, general), and CGI was already noticed at the end of first week. For the ESRS, parkinsonism rating scores were significantly increased until week 4 comparing with baseline. Dystonia rating scores were significantly increased until week 1, and dyskinesia rating scores were not significantly changed during the study. Laboratory parameters including vital sign, EKG, hematological, and biochemical values showed no significant changes during the trial. Conclusions : This study suggests that risperidone is generally safe and effective against both the positive and negative symptoms in our group of patients.

심인성 어지럼증 환자의 정신과적 증상에 관한 연구

이경규,이지영,김현우,이정엽,백기청,김재일,이근호 大韓神經精神醫學會 1999 신경정신의학 Vol.38 No.5

연구목적 : 어지럼증을 호소하는 환자들 중에 심인성 어지럼증 환자가 상당수 있다는 것은 알려져 있고, 이러한 심인성 어지럼증과 동반되는 정신질환으로는 불안장애, 우울장애, 인격장애 등이 있다. 이에 본 연구에서는 심인성 어지럼증 환자들의 빈도와 정신과적 증상을 알아보고자 하였다. 방 법 : 어지럼증 환자들에게 평형검사, 자세운동측정검사, 자세변화검사, caloric testing을 시행하여 심인성, 말초성 및 중추성의 3군으로 분류한 후 각각 14명, 16명, 32명을 대상으로 하였다. 대상환자들에게 한국판 Beck 우울척도검사(K-BDI), Spielberg의 상태-특성 불안검사(STAI). 간이정신진단검사(SCL-90-R)를 이용하여 정신과적 증상을 측정하여 이들 세 군을 비교분석하였다. 결 과 : 1) 심인성 어지럼증을 가진 환자는 어지럼증을 나타내는 환자 총 62명중에서 14명으로 22.6%를 차지하였다. 2) K-BDI에 따른 세 군간의 비교에서는 세 군간에 유의한 차이를 보이지 않았다. 3) STAI에 따른 세 군간의 비교에서 상태불안(STAI-S) 및 특성불안(STAI-T) 모두에서 세 군간에 유의한 차이를 보이지 않았다. 4) SCL-90-R에 따른 세 군간의 비교에서는 9개의 척도 중에서 공포불안척도에서만 중추성 어지럼증군이 47.56±6.90, 말초성 어지럼증 군이 53.50±13.74, 심인성 어지럼증 군이 58.50±16.05로 심인성 어지럼증 환자군이 중추성 어지럼증 환자군에 비하여 유의하게 높은 것으로 나타났다(p<0.01). 5) 통증 증상에 있어서는 세 군간에 유의한 차이를 보이지 않았고, 위장관 증상과 성기능 장애 증상은 중추성과 말초성 어지럼증 군이 심인성 어지럼증 군에 비하여 유의하게 높은 것으로 나타났다(p<0.01). 결 론 : 상기 결과를 토대로 했을 때 어지럼증 환자들에 대한 정신과적 접근을 위하여 각 군을 나누는 것은 별 의미가 없으며 어지럼증 환자들의 어지럼증 자체에 대한 직접적이고 집중적인 치료가 더욱 필요할 것으로 생각된다. Objectives : This study was aimed to investigate psychiatric symptoms in patients with psychogenic dizziness and compare these findings with those of patients with central and peripheral dizziness. Methods : A total of 62 patients with dizziness was the subject of investigation, and patients were classified into 32 with central type, 16 with peripheral type, and 14 with psychogenic type. Korean standardized Beck Depression Inventory(K-BDI), State and Trait Anxiety Inventory(STAI), and Korean standardized edition of Symptom Checklist 90 Revised(SCL-90-R) were used for the assessment. Statistically, Pearson's chi-square test and one-way ANOVA with Scheffe's test were used with SPSS/PC for windows 6.0. Results : The results were as follows : 1) The proportion of psychogenic dizziness was 22.6% of the total subjects. 2) Total scores of K-BDI were not significantly different among the 3 groups. 3) Total scores of state anxiety and trait anxiety were not significantly different among the 3 groups. 4) In SCL-90-R, psychogenic dizziness group showed significantly higher score of phobic anxiety only(p<0.01) than central dizziness group. And others were not significantly different among the 3 groups. 5) In additional somatic symptoms, pain score was not different among the 3 groups, but gastrointestinal and sexual symptoms scores of central and peripheral dizziness group were significantly higher than those of psychogenic dizziness group(p<0.01). Conclusion : These results suggest that psychiatric symptoms in patients with psychogenic dizziness are not different from those of patients with central or peripheral dizziness. Therefore, the more direct and intensive treatment may be necessary regardless of the type of dizziness.

Ethanol Extract of Ganoderma lucidum Augments Cellular Anti-oxidant Defense through Activation of Nrf2/HO-1

Lee, Yoo-hwan,Kim, Jung-hee,Song, Choon-ho,Jang, Kyung-jeon,kim, Cheol-hong,Kang, Ji-Sook,Choi, Yung-hyun,Yoon, Hyun-Min KOREAN PHARMACOPUNCTURE INSTITUTE 2016 Journal of pharmacopuncture Vol.19 No.1

Objectives: The mushroom Ganoderma lucidum has been widely used as a traditional herbal medicine for many years. Although several studies have focused on the anti-oxidative activity of this mushroom, the molecular mechanisms underlying its activity have not yet been clearly established. The present study investigated the cytoprotective effect of ethanol extract of Ganoderma lucidum (EGL) against oxidative stress (hydrogen peroxide, $H_2O_2$) and elucidated the underlying mechanisms in a C2C12 myoblast cell line. Methods: Oxidative stress markers were determined by using the comet assay to measure reactive oxygen species (ROS) generation and deoxyribonucleic acid (DNA) damage. Cell viability and Western blotting analyses were employed to evaluate the cellular response to EGL and $H_2O_2$ in C2C12 cells. Transfection with nuclear factor erythroid 2-related factor 2 (Nrf2)-specific small interfering ribonucleic acid (siRNA) was conducted to understand the relationship between Nrf2 expression and $H_2O_2$-induced growth inhibition. Results: The results showed that EGL effectively inhibited $H_2O_2$-induced growth and the generation of ROS. EGL markedly suppressed $H_2O_2$-induced comet-like DNA formation and phosphorylation of histone H2AX at serine 139 ($p-{\gamma}H2AX$), a widely used marker of DNA damage, suggesting that EGL prevented $H_2O_2$-induced DNA damage. Furthermore, the EGL treatment effectively induced the expression of Nrf2, as well as heme oxygenase-1 (HO-1), with parallel phosphorylation and nuclear translocation of Nrf2 in the C2C12 myoblasts. However, zinc protoporphyrin IX, a HO-1 inhibitor, significantly abolished the protective effects of EGL against $H_2O_2$-induced accumulation of ROS and reduced cell growth. Notably, transient transfection with Nrf2-specific siRNA attenuated the cytoprotective effects and HO-1 induction by EGL, indicating that EGL induced the expression of HO-1 in an Nrf2-dependent manner. Conclusion: Collectively, these results demonstrate that EGL augments the cellular anti-oxidant defense capacity through activation of Nrf2/HO-1, thereby protecting C2C12 myoblasts from $H_2O_2$-induced oxidative cytotoxicity.

Diatomaceous earth를 이용한 경제적이고 효율높은 homemade PCR purification column의 제조 및 평가

이동호,이승관,이창규,조경진,최명재,이국성 高麗大學校 倂設 保健大學 保健科學硏究所 2002 保健科學論集 Vol.28 No.2

In recent years the amount of sequencing has increased tremendously, especially due to the introduction of automated DNA sequencing. PCR is principal method for the synthesis of templates for DNA sequencing. Consequently, the need for PCR cleanup for template preparation has increased. Already, many PCR purification kits are marketed. This DNA purification kits uses most glass membrane or silica gel, So, can do rapidly efficient purification by simple miniprep. But, efficiency of purification kits is different according to manufacturer and is very expensive. Therefore, we compare the homemade Diatomaceous earth column with major manufacturer's spin column methods for DNA purification. As a result, we found that the homemade Diatomaceous earth column method resulted in equal or better yields relative to the commercial purification kits.