장기간 운동이 마우스 조직별 heme oxygenase-1 발현에 미치는 영향

김시영 ( Si Young Kim ) 한국운동생리학회(구-한국운동과학회) 2010 운동과학 Vol.19 No.1

Heme oxygenase-1 (HO-1)은 산화적인 자극을 포함한 다양한 외부 스트레스에 대한 조직세포의 적응 및 방어에 중요한 역할을 담당하는 항산화효소이다. 본 연구의 목적은 마우스의 다양한 조직에서 HO-1의 발현 차이는 물론 장기간의 운동에 의한 조절 가능성을 알아보는데 있다. 연구에 사용된 24마리의 ICR계 마우스는 각각 8마리씩 세 그룹 [control group (CONT, n=8), low-intensity exercise group (LIE, n=8), high-intensity exercise group (HIE, n=8)]으로 나누어 졌으며, 16주간의 트레드밀 운동프로그램 후 골격근, 대장, 심장, 폐, 간 그리고 뇌를 분리하여 분석하였다. CONT에서 안정시 마우스 조직별 HO-1 발현량을 분석한 결과, 간 조직에서의 발현량이 가장 높았으며, 이어서 대장, 폐, 심장, 골격근 그리고 뇌 순으로 발현량의 유의한 차이가 나타났다. 16주간 트레이닝 후 비교그룹 (CONT)에 비해 트레이닝 그룹 (HIE, LIE)의 골격근, 대장 및 간에서 HO-1의 발현량이 유의하게 증가된 (p<.01, p<.001) 반면, 심장, 폐 그리고 뇌 조직에서는 그룹 간 발현차이가 나타나지 않았다. 이상의 결과를 바탕으로 결론을 내리면, 안정된 상태에서 마우스 조직에 따라 발현되는 HO-1 단백질량은 조직 사이에 유의한 차이가 있으며, 골격근, 대장 그리고 간 조직에서의 HO-1 발현량은 반복된 운동으로 증가시킬 수 있었다. Heme oxygenase (HO)-1, involved in the heme degradation process, is upregulated by a wide array of stimuli and has antioxidant, anti-inflammatory and other cytoprotective functions. The aim of this study was to investigate the difference and effect of long-term exercise on HO-1 expression among mouse tissues. The study carried out on twenty-four ICR mice. All mice participated in this study which consisted of three groups [control group (CONT, n=8), low-intensity exercise group (LIE, n=8), high-intensity exercise group (HIE, n=8)]. Mice were euthanized after 16 weeks of training periods and tissues (skeletal muscle, colon, heart, lung, liver, and brain) were removed. As a result of analyzed difference of HO-1 expression among tissues, liver topped the list, followed by colon, lung, heart, skeletal muscle, and brain (p<.001). In case of skeletal muscle, colon, and liver, after training for 16 weeks, HO-1 expression in HIE and/or LIE dramatically increased in comparison with CONT (p<.01, p<.001). On the other hand, no significant differences in the heart, lung, and brain were found between all groups. Taken together, we conclude that level of HO-1 expression is different between mouse tissues and long-term regular exercise can enhance the levels of HO-1 expression in skeletal muscle, colon, and liver.

Defense mechanism of heme oxygenase-1 against cytotoxic and receptor activator of nuclear factor-&kgr;B ligand inducing effects of hydrogen peroxide in human periodontal ligament cells

Pi, S.-H.,Kim, S.-C.,Kim, H.-T.,Lee, H.-J.,Lee, S.-K.,Kim, E.-C. Blackwell Publishing Ltd 2007 Journal of periodontal research Vol.42 No.4

<P>Background and Objective: </P><P>Although induction of heme oxygenase-1 by H<SUB>2</SUB>O<SUB>2</SUB> has been reported, the protective role of heme oxygenase-1 against the cytotoxic and osteoclastogenic effects of H<SUB>2</SUB>O<SUB>2</SUB> have not been elucidated in human periodontal ligament cells. The aim of this work was to investigate the defense mechanism of heme oxygenase-1 on H<SUB>2</SUB>O<SUB>2</SUB>-induced cytotoxicity and to analyze the expression of receptor activator of nuclear factor-&kgr;B ligand (RANKL) and osteoprotegerin as markers for osteoclast differentiation in periodontal ligament cells.</P><P>Material and Methods: </P><P>Using human periodontal ligament cells, cytotoxicity was measured by the 3,4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) assay, and expression of heme oxygenase-1, RANKL, and osteoprotegerin mRNA was determined by reverse transcription–polymerase chain reaction.</P><P>Results: </P><P>H<SUB>2</SUB>O<SUB>2</SUB> produced a cytotoxic effect by reducing the cell viability and enhancing the expression of heme oxygenase-1 and RANKL mRNAs in a concentration- and time-dependent manner. Additional experiments revealed that heme oxygenase-1 inducer (hemin), a membrane-permeable cGMP analog (8-bromo-cGMP), carbon monoxide, extracellular signal-regulated kinase, p38 mitogen-activated protein kinase inhibitor, protein kinase inhibitor (KT5823), and nuclear factor-&kgr;B inhibitor (pyrrolidine dithiocarbamate) also blocked the effects of H<SUB>2</SUB>O<SUB>2</SUB> on cell viability and RANKL mRNA expression in periodontal ligament cells.</P><P>Conclusion: </P><P>These data suggest that heme oxygenase-1 induction plays a protective role in periodontal ligament cells against the cytotoxic and RANKL-inducing effects of H<SUB>2</SUB>O<SUB>2</SUB>, through multiple signaling pathways.</P>

Induction of Heme Oxygenase-1 Expression by Rapamycin and Paclitaxel in Smooth Muscle and Endothelial Cells of Human and Rat

Park, Chul Bo,Chung, Hun Taeg,Rhew, Hyun Yul 고신대학교 의학부 2005 高神大學校 醫學部 論文集 Vol.20 No.1

Background: Both rapamycin and paclitaxel are currently being used in drug-eluting stent to block the proliferation of vascular endothelial and smooth muscle cells. Heme oxygenase-1 (HO-1) has also known to be anti-proliferative against vascular cells. In this study, it focused on the role of HO-1 in cytoprotection and antiproliferation of paclirtaxel. Methods: Proliferation of cells and metabolic activity of HO-1 were evaluated with the tetrazolium-based assay. HO-1 protein expression was shown by western bolt analysis of vascular smooth muscle cells (VMSCs), or endothelial cells (ECs), using anti-HO-1 polyclonal antibody from untreated cells and cells exposed to rapamycin and paclitaxel. To determine whether the inhibition of paclitaxel on platelet-derived growth factor (PDGF)-detpendent proliferation may be mediated by its induction of HO-1, we exposed cells to paclitaxel, PDGF and ZnPP, an inhibitor of HO activity. Survival rate was checked to fine the anti-apototic effect of paclitaxel against tumor necrosis factor (TNF)-α-mediated apoptosis, in the presence or abscence of ZnPP. Result: Rapamycin induces HO-1 protein in rat VSMCs and human ECs. Exposure of VSMCs to palcitaxel for 12 h resulted in a dose-dependent increase in HO-1 activity. Ten nM of paclitaxel led to a maximal expression of HO-1 protein at 12h after exposure. The addition of Znp abrogated the suppression effect of paclitaxel on PDGF-stimulated cell proliferation, and blocked paclitaxel-mediated suppression of TNF-α mediated apoptosis. Hemoglobin, a scavenger of CO, abrogated the paclitaxel induced cytoprotection. Conclusion: Paclitaxel induced the expression of HO-1 gene in rat VSMCs and human ECs, in dose-dependent and time dependent manner, like rapamycin.

Oxymatrine inhibits the pyroptosis in rat insulinoma cells by affecting nuclear factor kappa B and nuclear factor (erythroid-derived 2)-like 2 protein/heme oxygenase-1 pathways

Jingying Gao,Lixia Xia,Yuanyuan Wei 대한약리학회 2022 The Korean Journal of Physiology & Pharmacology Vol.26 No.3

As the mechanism underlying glucose metabolism regulation by oxymatrine is unclear, this study investigated the effects of oxymatrine on pyroptosis in INS-1 cells. Flow cytometry was employed to examine cell pyroptosis and reactive oxygen species (ROS) production. Cell pyroptosis was also investigated via transmission electron microscopy and lactate dehydrogenase (LDH) release. Protein levels were detected using western blotting and interleukin (IL)-1β and IL-18 secretion by enzyme-linked immunosorbent assay. The caspase-1 activity and DNA-binding activity of nuclear factor kappa B (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2) were also assessed. In the high glucose and high fat-treated INS-1 cells (HG + PA), the caspase-1 activity and LDH content, as well as Nod-like receptor family pyrin domain containing 3, Gsdmd-N, caspase-1, apoptosis-associated specklike protein containing a CARD, IL-1β, and IL-18 levels were increased. Moreover, P65 protein levels increased in the nucleus but decreased in the cytoplasm. Oxymatrine attenuated these effects and suppressed high glucose and high fat-induced ROS production. The increased levels of nuclear Nrf2 and heme oxygenase-1 (HO-1) in the HG + PA cells were further elevated after oxymatrine treatment, whereas cytoplasmic Nrf2 and Keleh-like ECH-associated protein levels decreased. Additionally, the elevated transcriptional activity of p65 in HG + PA cells was reduced by oxymatrine, whereas that of Nrf2 increased. The results indicate that the inhibition of pyroptosis in INS- 1 cells by oxymatrine, a key factor in its glucose metabolism regulation, involves the suppression of the NF-κB pathway and activation of the Nrf2/HO-1 pathway.

Heme oxygenase-1 mediated protective effect of methyl gallate on cadmium-induced cytotoxicity in cultured mouse mesangial cells

Cha, Seok-Ho,Suh, Chang-Kook The Korean Society of Toxicogenomics and Toxicopro 2010 Molecular & cellular toxicology Vol.6 No.2

To clarify the effect of a phytochemical, methyl gallate (MG) on a heavy metal (cadmium)-induced renal toxicity, cytotoxicity and the change of heme oxygenase-1 (HO-1) gene expression was studied using cultured mouse renal glomerular mesangial cells (MMC). By employing RT-PCR and Western blotting analysis, we have examined the HO-1 induction in MMCs that were treated with $Cd^{2+}$ and/or MG. Using MTT assay we have also examined the cytoprotective effect of HO-1 induction against the cytotoxicity caused by toxic dose of $Cd^{2+}$. In MMCs exposed to $Cd^{2+}$ and MG, expression of HO-1 (mRNA and protein) was increased in a concentration- and time-dependent manner. The increments of HO-1 mRNA and protein expressions by $Cd^{2+}$ and MG were inhibited by the treatment of the cells with actinomycin D, an inhibitor of transcription. The decreased viability of the cells by $Cd^{2+}$ was partially recovered by the treatment of MG and this recovery by the MG was reduced by the treatment of zinc protoporphyrin IX (a HO-1 inhibitor). From these results, methyl gallate might have cytoprotective effect on $Cd^{2+}$-induced cytotoxicity that is related with heme oxygenase-1 induction.

15-Deoxy-Δ^12,14-prostaglandin J₂ induces p53 expression through Nrf2-mediated upregulation of heme oxygenase-1 in human breast cancer cells

Kim, D. H.,Song, N. Y.,Kim, E. H.,Na, H. K.,Joe, Y.,Chung, H. T.,Surh, Y. J. Informa Healthcare 2014 Free radical research Vol.48 No.9

<P>Heme oxygenase-1 (HO-1) is a stress-responsive enzyme that has antioxidant and cytoprotective functions. However, HO-1 has oncogenic functions in cancerous or transformed cells. In the present work, we investigated the effects of HO-1 on the expression of p53 induced by 15-deoxy-Δ<SUP>12,14</SUP>-prostaglandin J<SUB>2</SUB> (15d-PGJ<SUB>2</SUB>) in human breast cancer (MCF-7) cells. Treatment of MCF-7 cells with 15d-PGJ<SUB>2</SUB> led to time-dependent increases in the expression of p53 as well as HO-1. Upregulation of p53 expression by 15d-PGJ<SUB>2</SUB> was abrogated by si-RNA knock-down of HO-1. In MCF-7 cells transfected with HO-1 si-RNA, 15d-PGJ<SUB>2</SUB> failed to induce expression of p53 as well as HO-1. In addition, HO-1 inducers enhanced the p53 expression. We speculated that iron, a by-product of HO-1-catalyzed reactions, could mediate 15d-PGJ<SUB>2</SUB>-induced p53 expression. Upregulation of p53 expression by 15d-PGJ<SUB>2</SUB> was abrogated by the iron chelator desferrioxamine in MCF-7 cells. Iron released from heme by HO-1 activity is mostly in the Fe<SUP>2+</SUP> form. When MCF-7 cells were treated with the Fe<SUP>2+</SUP>-specific chelator phenanthroline, 15d-PGJ<SUB>2</SUB>-induced p53 expression was attenuated. In addition, levels of the Fe-sequestering protein H-ferritin were elevated in 15d-PGJ<SUB>2</SUB>-treated MCF-7 cells. In conclusion, upregulation of p53 and p21 via HO-1 induction and subsequent release of iron with accumulation of H-ferritin may confer resistance to oxidative damage in cancer cells frequently challenged by redox-cycling anticancer drugs.</P>

폐암세포주에서 Heme Oxygenase-1의 억제가 Cisplatin의 항암제 감수성에 미치는 영향

김소영 ( So Young Kim ),김은정 ( Eun Jung Kim ),장혜연 ( Hye Yeon Jang ),황기은 ( Ki Eun Hwang ),박정현 ( Jung Hyun Park ),김휘정 ( Hwi Jung Kim ),조향정 ( Hyang Jeong Jo ),양세훈 ( Sei Hoon Yang ),정은택 ( Eun Taik Jeong ),김학렬 대한결핵 및 호흡기학회 2007 Tuberculosis and Respiratory Diseases Vol.62 No.1

연구배경: 다양한 고형암에서 HO-1의 높은 발현이 알려져 있고, 그것의 항산화와 항세포고사의 역할로 인해 빠른 암종의 성장에 중요한 역할이 있음이 보고되고 있다. 대표적인 활성산소종 생성 항암제인 Cisplatin은 현재까지 폐암치료에 가장 광범위하게 사용되고 있으나, 여러 내성발생이 임상치료의 주요문제로 대두되고 있다. 저자들은 A549 폐암세포주에서 HO-1의 발현이 증가되었고 HO-1 활성억제제나 siRNA 방법을 통해 생존율의 의미 있는 감소와 세포고사가 유도됨을 보고한 바 있다. 이 연구의 목적은 A549 폐암세포주에 cisplatin 처리시 HO-1의 발현의 증가유무와 기전을 규명하고 실제 HO-1의 억제가 cisplatin에 의한 항암제 감수성을 증가시키는지를 알아보는데 있다. 방법: 비소세포폐암세포주인 A549, NCI-H23, NCI-H157, NCI-H460을 이용하였다. 세포독성은 MTT 방법으로 구하였고, HO-1, Nrf2, MAPK의 발현은 Western blotting으로 확인하였다. 또한 MAPK억제제들을 전처치한 후 cisplatin에 의해 유도된 Nrf2와 HO-1의 발현에 미치는 영향을 역시 Western blotting으로 관찰하였다. A549세포에 활성억제제인 ZnPP나 HO-1 small interfering RNA (siRNA)을 주입하여 cisplatin과의 병합요법시 생존율의 배가효과 유무를 MTT 방법으로 확인하였고, 이러한 효과가 ROS 형성과 HO-1의 발현변화와 관련되는지를 알아보기 위해 carboxy-H2DCFDA 방법과 Western blotting을 통해 각각 확인하였다. 결과: Cisplatin 처리시 다른 세포주에 비해 A549세포가 의의 있게 내성을 보였다. 10μM의 농도에서 시간 의존적으로 HO-1, Nrf2, MAPK의 발현이 증가하였고, MAPK 억제제들을 전 처치하였을 때 cisplatin에 의해 유도된 HO-1과 Nrf2의 발현이 억제됨을 확인하였다. HO-1의 활성억제제인 ZnPP와 HO-1 siRNA를 통해 HO-1 mRNA를 직접 억제하는 방법으로 cisplatin과 병합치료시 단독치료에 비해 의의 있는 생존율의 감소를 보였다. 이러한 효과는 활성산소종의 생성 증가와 HO-1의 발현억제에 의한 결과임을 확인하였다. 결론: Cisplatin 처리시 HO-1의 발현은 MAPK-Nrf2-HO-1의 경로를 통해 증가하였고, 부분적으로 치료에 대한 내성과 관련이 있었으며, ZnPP 등의 활성억제제나 siRNA를 통한 knock-down 방법으로 HO-1을 표적으로 억제하는 치료방법을 통해 cisplatin의 항암제 감수성을 증가시켰다. Background: Heme oxygenase-1 (HO-1) is known to modulates the cellular functions, including cell proliferation and apoptosis. It is known that a high level of HO-1 expression is found in many tumors, and HO-1 plays an important role in rapid tumor growth on account of its antioxidant and antiapoptotic effects. Cisplatin is a widely used anti-cancer agent for the treatment of lung cancer. However, the development of resistance to cisplatin is a major obstacle to its use in clinical treatment. We previously demonstrated that inhibiting HO-1 expression through the transcriptional activation of Nrf2 induces apoptosis in A549 cells. The aim of this study was to determine of the inhibiting HO-1 enhance the chemosensitivity of A549 cells to cisplatin. Materials and Methods: The human lung cancer cell line, A549, was treated cisplatin, and the cell viability was measured by a MTT assay. The change in HO-1, Nrf2, and MAPK expression after the cisplatin treatment was examined by Western blotting. HO-1 inhibition was suppressed by ZnPP, which is a specific pharmacologic inhibitor of HO activity, and small interfering RNA (siRNA). Flow cytometry analysis and Western blot were performed in to determine the level of apoptosis. The level of hydrogen peroxide (H2O2) generation was monitored fluoimetrically using 2`,7`-dichlorofluorescein diacetate. Results: The A549 cells showed more resistance to the cisplatin treatment than the other cell lines examined, whereas cisplatin increased the expression of HO-1 and Nrf2, as well as the phosphorylation of MAPK in a time-dependent fashion. Inhibitors of the MAPK pathway blocked the induction of HO-1 and Nrf2 by the cisplatin treatment in A549 cells. In addition, the cisplatin-treated A549 cells transfected with dither the HO-1 small interfering RNA (siRNA) or ZnPP, specific HO-1 inhibitor, showed in a more significantly decrease in viability than the cisplatin-only-treated group. The combination treatment of ZnPP and cisplatin caused in a marked increase in the ROS generation and a decrease in the HO-1 expression. Conclusion: Cisplatin increases the expression of HO-1, probably through the MAPK-Nrf2 pathway, and the inhibition of HO-1 enhances the chemosensitivity of A549 cells to cisplatin. (Tuberc Respir Dis 2007; 62: 33-42)

Non-canonical vs. Canonical Functions of Heme Oxygenase-1 in Cancer

Jagadeesh Achanta Sri Venakata,Fang Xizhu,김성훈,Guillen-Quispe Yanymee N.,Zheng Jie,서영준,Kim Su-Jung 대한암예방학회 2022 Journal of cancer prevention Vol.27 No.1

Heme oxygenase-1 (HO-1) is a critical stress-responsive enzyme that has antioxidant and anti-inflammatory functions. HO-1 catalyzes heme degradation, which gives rise to the formation of carbon monoxide (CO), biliverdin, and iron. The upregulation of HO-1 under pathological conditions associated with cellular stress represents an important cytoprotective defense mechanism by virtue of the anti-oxidant properties of the bilirubin and the anti-inflammatory effect of the CO produced. The same mechanism is hijacked by premalignant and cancerous cells. In recent years, however, there has been accumulating evidence supporting that the upregulation of HO-1 promotes cancer progression, independently of its catalytic activity. Such non-canonical functions of HO-1 are associated with its interaction with other proteins, particularly transcription factors. HO-1 also undergoes post-translational modifications that influence its stability, functional activity, cellular translocation, etc. HO-1 is normally present in the endoplasmic reticulum, but distinct subcellular localizations, especially in the nucleus, are observed in multiple cancers. The nuclear HO-1 modulates the activation of various transcription factors, which does not appear to be mediated by carbon monoxide and iron. This commentary summarizes the non-canonical functions of HO-1 in the context of cancer growth and progression and underlying regulatory mechanisms.

15d-PGJ2 inhibits NF-kB and AP-1-mediated MMP-9 expression and invasion of breast cancer cell by means of a heme oxygenase-1-dependent mechanism

장혜연,On-Yu Hong,Hyun Jo Youn,김민걸,Cheorl-Ho Kim,정성후,Jong-Suk Kim 생화학분자생물학회 2020 BMB Reports Vol.53 No.4

Activation of peroxisome proliferator-activated receptor  (PPAR) serves as a key factor in the proliferation and invasion of breast cancer cells and is a potential therapeutic target for breast cancer. However, the mechanisms underlying this effect remain largely unknown. Heme oxygenase-1 (HO-1) is induced and overexpressed in various cancers and is associated with features of tumor aggressiveness. Recent studies have shown that HO-1 is a major downstream target of PPAR. In this study, we investigated the effects of induction of HO-1 by PPAR on TPAinduced MMP-9 expression and cell invasion using MCF-7 breast cancer cells. TPA treatment increased NF-B /AP-1 DNA binding as well as MMP-9 expression. These effects were significantly blocked by 15d-PGJ2, a natural PPAR ligand. 15d-PGJ2 induced HO-1 expression in a dose-dependent manner. Interestingly, HO-1 siRNA significantly attenuated the inhibition of TPA-induced MMP-9 protein expression and cell invasion by 15d-PGJ2. These results suggest that 15d-PGJ2 inhibits TPA-induced MMP- 9 expression and invasion of MCF-7 cells by means of a heme oxygenase-1-dependent mechanism. Therefore, PPAR/HO-1 signaling- pathway inhibition may be beneficial for prevention and treatment of breast cancer.

Delivery of Hypoxia Inducible Heme Oxygenase-1 Gene Using Dexamethasone Conjugated Polyethylenimine for Protection of Cardiomyocytes under Hypoxia

Kim, Hyun-Jung,Kim, Hyun-Ah,Choi, Joon-Sig,Lee, Min-Hyung Korean Chemical Society 2009 Bulletin of the Korean Chemical Society Vol.30 No.4

Heme oxygenase-1 (HO-1) is an anti-inflammatory and anti-apoptotic protein and has been applied to various gene therapy researches. However, constitutive expression of HO-1 may induce deleterious side effects. In this research, hypoxia inducible HO-1 expression plasmid, pEpo-SV-HO-1, was constructed with the erythropoietin (epo) enhancer and simian virus 40 (SV40) promoter to avoid these unwanted side effects. Dexamethasone conjugated polyethylenimine (PEI-Dexa) was used as a gene carrier. It was previously reported that dexamethasone protected cardiomyocytes from apoptosis under hypoxia. In this research, PEI-Dexa reduced the caspase-3 level in hypoxic H9C2 cardiomyocytes as a derivative of dexamethasone, suggesting that PEI-Dexa is an anti-apoptotic reagent as well as a gene carrier. pEpo-SV-HO-1 was transfected to H9C2 cardiomyocytes using PEI-Dexa and the cells were incubated under normoxia or hypoxia. HO-1 expression was induced in the pEpo-SV-HO-1 transfected cells under hypoxia. In addition, cell viability under hypoxia was higher in the pEpo-SV-HO-1 transfected cells than the pEpo-SV-Luc transfected cells. Also, caspase-3 level was reduced in the pEpo-SV-HO-1 transfected cells under hypoxia. In addition to the anti-apoptotic effect of PEI-Dexa, hypoxia inducible HO-1 expression by pEpo-SVHO- 1 may be helpful to protect cardiomyocytes under hypoxia. Therefore, pEpo-SV-HO-1/PEI-Dexa complex may be useful for ischemic heart disease gene therapy.