PGA2 induces the expression of HO-1 by activating p53 in HCT116 cells

Hyein Lee,Sang-Sun Lee,Ji-Young Park,Yun-Jeong Choe,이선영,Ho-Shik Kim,H.-S. Kim 대한독성 유전단백체 학회 2017 Molecular & cellular toxicology Vol.13 No.2

Prostaglandin (PG) A2 which is a cytotoxic PG, was reported to induce the expression of heme oxygenase (HO)-1 via activation of p38MAPK to keep U2OS cells from cell cycle arrest in G2M phase. The expression of HO-1 is primarily regulated at the level of transcription. But the transcription factors that are responsible for PGA2-induced HO-1 expression were not clarified yet. Here, we report that PGA2-induced transcription of HO-1 is mediated by p53, a tumor suppressive transcription factor. In HCT116 cells, PGA2 treatment led to the phosphorylation of p53 and an increase of p21WAF1 transcription as well as the activation of HO-1 transcription. Knocking p53 down via RNA interference or inhibiting the p53’s transcriptional activity by pifithrin-α treatment led to suppression of the increase in the level of both HO-1 expression and activity of HO-1 promoter. Pretreatment of NU- 7441, a chemical inhibitor of DNA-activated protein kinase (DNA-PK), prevented both the PGA2-induced phosphorylation of p53 and an increase of HO-1 transcription. In addition, N-acetyl-l-cysteine, a scavenger of reactive oxygen species (ROS), also mimicked the effect of NU-7441 on the PGA2-induced activation of p53 and HO-1 transcription. Collectively, these results suggest that PGA2 induces the expression of HO-1 via activation of p53, which is mediated by the ROSDNA- PK pathway.

Hypoxia-Responsive MicroRNA-101 Promotes Angiogenesis <i>via</i> Heme Oxygenase-1/Vascular Endothelial Growth Factor Axis by Targeting Cullin 3

Kim, Ji-Hee,Lee, Kwang-Soon,Lee, Dong-Keon,Kim, Joohwan,Kwak, Su-Nam,Ha, Kwon-Soo,Choe, Jongseon,Won, Moo-Ho,Cho, Byung-Ryul,Jeoung, Dooil,Lee, Hansoo,Kwon, Young-Guen,Kim, Young-Myeong Mary Ann Liebert 2014 Antioxidants & redox signaling Vol.21 No.18

<P>Aims: Hypoxia induces expression of various genes and microRNAs (miRs) that regulate angiogenesis and vascular function. In this study, we investigated a new functional role of new hypoxia-responsive miR-101 in angiogenesis and its underlying mechanism for regulating heme oxygenase-1 (HO-1) and vascular endothelial growth factor (VEGF) expression. Results: We found that hypoxia induced miR-101, which binds to the 3 ' untranslated region of cullin 3 (Cul3) and stabilizes nuclear factor erythroid-derived 2-related factor 2 (Nrf2) via inhibition of the proteasomal degradation pathway. miR-101 overexpression promoted Nrf2 nuclear accumulation, which was accompanied with increases in HO-1 induction, VEGF expression, and endothelial nitric oxide synthase (eNOS)-derived nitric oxide (NO) production. The elevated NO-induced S-nitrosylation of Kelch-like ECH-associated protein 1 and subsequent induction of Nrf2-dependent HO-1 lead to further elevation of VEGF production via a positive feedback loop between the Nrf2/HO-1 and VEGF/eNOS axes. Moreover, miR-101 promoted angiogenic signals and angiogenesis both in vitro and in vivo, and these events were attenuated by inhibiting the biological activity of HO-1, VEGF, or eNOS. Moreover, these effects were also observed in aortic rings from HO-1(+/-) and eNOS(-/-) mice. Local overexpression of miR-101 improved therapeutic angiogenesis and perfusion recovery in the ischemic mouse hindlimb, whereas antagomiR-101 diminished regional blood flow. Innovation: Hypoxia-responsive miR-101 stimulates angiogenesis by activating the HO-1/VEGF/eNOS axis via Cul3 targeting. Thus, miR-101 is a novel angiomir. Conclusion: Our results provide new mechanistic insights into a functional role of miR-101 as a potential therapeutic target in angiogenesis and vascular remodeling. Antioxid. Redox Signal. 21, 2469-2482.</P>

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]

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.

부산·울산·경남지역 직업병 감시체계

김정일,김병권,김정원,채창호,이철호,강동묵,김지홍,김진하,김영욱,이영하,이지호,김정호,윤형렬,유철인,정백근,장태원,김운규,윤동영,강진욱,김종은,안진홍,이동준,장준호,이광영,송혜란,최영희,이용환,조병만,최홍렬,고상백,김은아,이유진,홍영습,정갑열,김정만,김준연 大韓産業醫學會 2004 대한직업환경의학회지 Vol.16 No.1

Objective: Occupational medicine specialists in the Busan, Ulsan and Kyung-Nam areas established an area-based occupational disease surveillance system and used this system to collected case information for the purpose of preventing occupational diseases Methods: l l hospital participated in this system. The authors selected five main diseases, which were hand-arm vibration syndrome (HAVS), work-related musculoskeletal disorder (WRMSD), occupational asthma, occupational skin disease and occupational and occupational lung cancer and established their case definitions. All cases were reported on the web, and real time analysis was conducted. Results: Between April 2001 and April 2003, 192 cases of HAVS, 118 cases of WRMSD, 33 cases of occupational asthma, 17 cases of occupational skin disease and 17 cases of occupational lung cancer (for a total of 377 cases of these five main diseases) were reported. most of the HAVS cases came from shipbuilding, and 172 of them (92.7%) were associated with grinding. Of the four main types of WRMSD, carpal tunnel syndrome (CTS) was the most prevalent with 46 cases and shipbuilding was also the main industry involved (83.9%). The ergonomic risk factors involved mainly associated with the hand. In 19 (57.6%) and 4 (12.1%) cases of occupational asthma, the agents involved. The causative agents of occupational lung cancer included Cr, welding fume, PAH, Ni, etc. 10(58.8%) of the cases were diagnosed as squamous cell carcinoma and 5(29.4%) as adenoma. Conclusion: This result showed that an area-based occupational disease surveillance system might provide an effective method of evaluating the prevalence of such diseases, however the Busan, Ulsan and Kyung-Nam provinces are too large to be treated as individual areas. Therefore, the authors suggest that each province should establish its own surveillance system.

Induction of heme oxygenase-1 protects against podocyte apoptosis under diabetic conditions

Lee, Sang Choel,Han, Seung Hyeok,Li, Jin Ji,Lee, Sun Ha,Jung, Dong-Sub,Kwak, Seung-Jae,Kim, Seung Hye,Kim, Dong Ki,Yoo, Tae-Hyun,Kim, Jin Hyun,Chang, Se-Ho,Han, Dae Suk,Kang, Shin-Wook International Society of Nephrology 2009 Kidney international Vol.76 No.8

Heme oxygenase-1 (HO-1) is an anti-oxidant enzyme normally upregulated in response to oxidant injury. Here we determined the role of HO-1 in podocyte apoptosis in glomeruli of streptozotocin-treated rats and in immortalized mouse podocytes cultured in media containing normal or high glucose. HO-1 expression, its activity, the ratio of Bax/Bcl-2 protein, and active caspase-3 fragments were all significantly higher in isolated glomeruli of diabetic rats and in high glucose–treated podocytes. These increases were inhibited by zinc protoporphyrin treatment of the rats or by HO-1 siRNA treatment of the podocytes in culture. The number of apoptotic cells was also significantly increased in the glomeruli of diabetic rats and in high glucose–treated podocytes. Inhibition of HO-1 accentuated the increase in apoptotic cells both in vivo and in vitro. Our findings suggest that HO-1 expression protects against podocyte apoptosis under diabetic conditions.

Sarcomatoid Carcinoma Arising from Mature Cystic Teratoma

Ho-Chang Lee1, Seung-Myoung Son, Yong-Moon Lee, Ji Hae Koo, Song-Yi Choi, Ok-Jun Lee, Eun-Hwan Jeong 충북대학교 동물의학연구소 2012 Journal of Biomedical and Translational Research Vol.13 No.2

Malignant transformation of mature cystic teratoma (MCT) is rare. Sarcomatoid carcinoma is a neoplasm comprising malignant mesenchymal cells and a conventional carcinomatous area. Here, we report on a case of sarcomatoid carcinoma arising from an MCT in the left ovary of a 45-year-old female. A unilocular cyst consistent with MCT was observed; however, a nodule within the cyst was confirmed from the resected ovary. Microscopically, the nodule showed both squamous cell carcinoma and pleomorphic sarcomatous components admixing with each other. Lining epithelial cells at the periphery of the main tumor showed squamous metaplasia. When a sarcomatous component is observed in the ovary tumor, it is important to find a squamous cell component, either benign or malignant.

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.

울산 석유화학공단 인근 어린이들의 혈액상변화 및 면역기능

이충렬,유철인,이지호,이헌,김양호 대한산업의학회 2001 대한직업환경의학회지 Vol.13 No.2

목적 : 지역 특성상 휘발성 유기화합물, 분진, 악취 등 환경오염의 영향을 많이 받는 것으로 추정되는 울산석유화학공단 인근 어린이들의 혈액상 변화, 계절 차이에 의한 혈액상 변화와 면역기능에 미치는 영향을 평가하기 위하여 본 연구를 실시하였다. 방법 : 2000년 4월, 7월, 10월 3회에 걸쳐 석유화학공단 인근 A 초등학생 46명(남자 32명, 여자 14명), B 초등학생 97명(남자 49명, 여자 48명)과 교외지역의 A 초등학생 95명(남자 47명, 여자 48명)을 대상으로 일반혈액검사, 백혈구 백분율 검사를 실시하였으며 7월 조사 때는 B 및 C 초등학생 11세 남자 각각 25명을 대상으로 면역학적 기능검사를 실시하였다. 결과 : 3차례에 걸쳐 진행된 검사결과는 계절별로 다른 양상을 보여 주었다. 즉, 총백혈구수, 호중구수, 림프구수는 A와 C 초등학생은 7월 조사에서는 감소되었다 다시 10월 조사 때는 다시 증가하는 경향을 보여주며, B초등학생은 계절별 변화가 거의 없는 것으로 나타났다. 적혈구수는 B와 C 초등학생은 7월 조사시 감소되었다가 10월 조사 때는 다시 증가하는 양상을 보여주나 A 초등학생은 갈수록 감소하는 경향을 보여주었다. 혈색소량의 변화는 적혈구수의 변화와 유사하였다 혈소판수는 모두 7월 조사 때는 감 소되었다가 10월 조사 때는 다시 증가하는 양상을 보여주었다. 조사시기에 따라 각 학교 어린이들간에 혈구세포수의 차이가 있음을 구명하기 위하여 generalized linear model의 repeated measures ANOVA를 시도한 결과 연령과 성을 공변인으로 보정한 후의 P 값에서 조사시기의 차이와 학교간 차이의 교호작용변수(interaction variable)는 모든 혈구세포에서 유의한 것으로 나타났다(P<0.05). 이는 조사시기에 따라 각 학교어린이들간에 혈구세포수의 차이가 유의하게 다르다는 것을 의미한다. 면역기능 평가를 위하여 혈중 immunoglobulin 측정 및 림프구 아형 분포분석을 B 및 C 초등학생 6학년 남학생 각 25명, 총 50명을 대상으로 실시하였으나 모두 정상범위이었고 학교간 차이도 없었다. 결론 : 휘발성 유기화합물 같은 물질에 쉽게 영향을 받을 수 있는 백혈구와 적혈구 등의 혈구세포는 조사시기에 따라 각 학교어린이들간에 혈구세포수의 차이가 유의하게 나타남은 각 학교 어린이들간에 조사시기에 따라 혈구세포수에 영향을 주는 휘발성 유기화합물의 노출량의 차이가 중요한 원인 중 하나일 것으로 추정된다. 향후 혈구세포의 변화의 원인이라 추정되는 휘발성 유기화합물의 기중 농도측정이나 체내 생체지표검사를 통한 노출원 조사를 병행하는 추가적 조사가 필요한 것으로 사료된다. Objectives : To evaluate the hematological changes and the immunological function of children living near the petrochemical estate in Ulsan. Methods : The study subjects comprised of 238 children who consisted of 143 children living near the petrochemical estate and 95 children living in a suburban area. We conducted the hematological examination 3 times, in April, July and October. Also we evaluated the immunological function of some children in July. To confirm differences between schools repeated measures ANOVA of generalized linear model was done controlling age and sex as covariates. Results : The blood cell counts of children showed differences between schools in accordance with the survey month. The total WBC, neutrophil, and lymphocyte counts of children living in a suburban area decreased in July and increased in October again, but those of some children living near the petrochemical estate did not changed distinctly. The RBC counts of children living in a suburban area decreased in July and increased in October again, but those of some children living near the petrochemical estate decreased as time passed. The changes of the amount of hemoglobin of all study subjects were similar with those of RBC. The platelet counts of all study subjects decreased in July and increased in October again. In the generalized linear model, school was a significant independent variable for the total WBC, RBC, and platelet counts, and sex was a significant independent variable for the RBC counts. Age was a significant independent variable for the iymphocyte and platelet counts. P values of all blood cell were statistically significant in interaction variable between the survey month and school, and those of the total WBC, neutrophil, and iymphocyte counts were statistically significant in interaction variable between the survey month and age. The immunological function showed no significant difference between study groups. Conclusions : The total WBC and RBC counts which would be easily affected by volatile organic compounds (VOCs) in the atmosphere showed differences between schools in accordance with the survey month. This suggests that the amounts of exposure to VOCs are different among each school children in different months. To ascertain the hematological changes by VOCs, an additional study evaluating the concentrations of atmosphere of VOCs and biological monitoring of some VOCs is needed.

유기용제노출이 직업적 청력손실에 미치는 영향

이지호,고영주,이헌,강정학,유철인,이충렬,김양호 大韓産業醫學會 2000 대한직업환경의학회지 Vol.12 No.4

목적 : 산업 장에서 노출되는 유기용제가 청력역치에 어떠한 영향을 미치는가를 5년 동안의 추적조사를 통하여 파악하고자 하였다. 방 법 : 유기용제와 소음에 동시에 노출된 근로자 43명과 소음노출수준, 연령, 근무경력을 짝짓기 한소음노출군을 대상으로 문진과 이경검사, 소음노출 수준, 유기용제노출수준을 조사하였다. 결과 : 1. 전체 근로자의 각 주파수별 청력역치 변화양상에서 500Hz∼2000Hz에서는 추적기간 동 소음동안 현저한 변화가 없었고, 4000Hz 및 8000Hz에서는 역치가 증가하는 경향을 나타내었다. 2.유기용제와 소음에 동시에 노출된군에서 소음 노출군에 비해 4000Hz 이상에서 청력역치가 높게 나타났으나, 통계적인 유의성은 없었다(p(0.05). 3.반복측정 분산분석 법으로 각 주파수별 평균청력 역치의 연차적 변화에 미치는 영향을 분석한 결과 유기용제 노출은 통계적으로 유의한 영향이 없었으나(p)0.05), 소음노출수준은 250Hz, 2000Hz 및 4000Hz에서, 연령은 250Hz와 500Hz에서 통계학적으로 유의한 영향을 미치는 것으로 나타났다(p(0.05). 4.소음노출군파 동시노출군 각각에서의 청력역치 변화량을 paired t-test로 비교한 결과 250Hz∼4000Hz에서는 통계학적 유의성이 없었으며(p(0.1), 8000Hz에서만 경계치 (p=0.0884)의 유의성을 나타내었다. 5.요인들의 영향을 보정한 상태에서 관찰하기 위해 관찰기간동안의 청력역치변화량을 종속변수로 두고 일반선형모형을 이용하여 주파수별로 관찰한 결과 8000Hz에서만 모델에 통계학적 유의성이 있었으며 (p(0.05), 여기서 소음노출수준이 통계학적으로 유의한 영향을 미치는 것으로 나타났고(p(0.05), 유기용제노출은 경 계수준이 었다(p=0.087). 결론 : 청력에 대한 유기용제의 영향이 소음과 연령에 비해 상대적으로 미미하여 명확한 결론에 이르지는 못하였으나, 다소 영향이 있는 것으로 생각되어 유기용제의 영향에 대한 추가연구가 필요할 것으로 판단된다. 또한 보다 효과적인 근로자들의 청력관리를 위해서는 소음 외 산업 장에서 동시에 노출되는 여러 유해물질들을 고려하여야 할 것이다. Objectives : This study was to evaluate the impacts of simultaneous exposure to noise and mixed solvents on workers'hearing threshold level (HTL) over five-year period. Methods : The study was conducted by interview and annual audiometric test on workers in ship building industry from 1994 to 1998. The cases(workers who exposed to noise and mixed solvent simultaneously) were 43 male workers and controls (workers who exposed to nolle) were selected by matching method with regard to age, sex, carrier, and noise exposure level. To assess the impacts of solvent exposure on hearing threshold level, with considering other factors, general liner model was used. Results : 1. The audiogram of all subjects showed typical sensorineural hearing loss pattern. The mean HTLe were increased at high frequency (4000HB and 8000Hz) for study period. 2. The HTLs of cases were more increased than those of controls at high frequencies (4000Hz and 8000Hz), but there was not statlstical significance. 3. The impact of age on the HTL was statistically significant at 250Hz and 500Hz (p(0.05), and the impact of noise on the HTL was statistically significant at 250Hz, 2000Hz and 4000Hz (p(0.05), but the impact solvent exposure on the HTL was not significant. 4. The changes in HTLs of cases were higher than those of controls at high frequency, there were not statistical significance at 4000Hz, but only at 8000Hz (p=0.087). 5. Statistical analysis of the general linear model implicated that the changes in HTL was impacted by noise exposure bevel significantly (p=0.031) and Impacted by solvent exposure weakly (p=0.087) at 8000Hz. Conclusions : The results of this study suggest that workers who simultaneously exposed to noise and mixed solvent were at risk for more affected HTL than those exposed to noise exposure only, but we could not find definitive evidence. Further detailed studies must be made in large number of workers.