Heme oxygenase-1 (HO-1)/carbon monoxide (CO) axis suppresses RANKL-induced osteoclastic differentiation by inhibiting redox-sensitive NF-κB activation

( Sun-uk Bak ),( Suji Kim ),( Hae-jun Hwang ),( Jung-a Yun ),( Wan-sung Kim ),( Moo-ho Won ),( Ji-yoon Kim ),( Kwon-soo Ha ),( Young-guen Kwon ),( Young-myeong Kim ) 생화학분자생물학회(구 한국생화학분자생물학회) 2017 BMB Reports Vol.50 No.2

Heme oxygenase (HO-1) catalyzes heme to carbon monoxide (CO), biliverdin/bilirubin, and iron and is known to prevent the pathogenesis of several human diseases. We assessed the beneficial effect of heme degradation products on osteoclastogenesis induced by receptor activator of NF-κB ligand (RANKL). Treatment of RAW264.7 cells with CORM-2 (a CO donor) and bilirubin, but not with iron, decreased RANKLinduced osteoclastogenesis, with CORM-2 having a more potent anti-osteogenic effect. CORM-2 also inhibited RANKLinduced osteoclastogenesis and osteoclastic resorption activity in marrow-derived macrophages. Treatment with hemin, a HO-1 inducer, strongly inhibited RANKL-induced osteoclastogenesis in wild-type macrophages, but was ineffective in HO-1^+/-cells. CORM-2 reduced RANKL-induced NFATc1 expression by inhibiting IKK-dependent NF-κB activation and reactive oxygen species production. These results suggest that CO potently inhibits RANKL-induced osteoclastogenesis by inhibiting redox-sensitive NF-κB-mediated NFATc1 expression. Our findings indicate that HO-1/CO can act as an antiresorption agent and reduce bone loss by blocking osteoclast differentiation. [BMB Reports 2017; 50(2): 103-108]

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>

Upregulation of heme oxygenase-1 by ginsenoside Ro attenuates lipopolysaccharide-induced inflammation in macrophage cells

Kim, Sokho,Oh, Myung-Hoon,Kim, Bum-Seok,Kim, Won-Il,Cho, Ho-Seong,Park, Byoung-Yong,Park, Chul,Shin, Gee-Wook,Kwon, Jungkee The Korean Society of Ginseng 2015 Journal of Ginseng Research Vol.39 No.4

Background: The beneficial effects of ginsenoside species have been well demonstrated in a number of studies. However, the function of ginsenoside Ro (GRo), an oleanane-type saponin, has not been sufficiently investigated. Thus, the aim of the present study was to investigate the anti-inflammatory effects of GRo in vitro using the Raw 264.7 mouse macrophage cell line treated with lipopolysaccharide (LPS), and to clarify the possible mechanism of GRo involving heme oxygenase-1 (HO-1), which itself plays a critical role in self-defense in the presence of inflammatory stress. Methods: Raw 264.7 cells were pretreated with GRo (up to $200{\mu}M$) for 1 h before treatment with 1 mg/mL LPS, and both cell viability and inflammatory markers involving HO-1 were evaluated. Results: GRo significantly increased cell viability in a dose dependent manner following treatment with LPS, and decreased levels of reactive oxygen species and nitric oxide. GRo decreased inflammatory cytokines such as nitric oxide synthase and cyclooxygenase-2 induced by LPS. Moreover, GRo increased the expression of HO-1 in a dose dependent manner. Cotreatment of GRo with tin protoporphyrin IX, a selective inhibitor of HO-1, not only inhibited upregulation of HO-1 induced by GRo, but also reversed the anti-inflammatory effect of GRo in LPS treated Raw 264.7 cells. Conclusion: GRo induces anti-inflammatory effects following treatment with LPS via upregulation of HO-1.

Korean Red ginseng prevents endothelial senescence by downregulating the HO-1/NF-κB/miRNA-155-5p/eNOS pathway

Tae-Hoon Kim,Ji-Yoon Kim,Jieun Bae,Young-Mi Kim,Moo-Ho Won,Kwon-Soo Ha,Young-Guen Kwon,Young-Myeong Kim 고려인삼학회 2021 Journal of Ginseng Research Vol.45 No.2

Background: Korean Red ginseng extract (KRGE) has beneficial effects on the cardiovascular system by improving endothelial cell function. However, its pharmacological effect on endothelial cell senescence has not been clearly elucidated. Therefore, we examined the effect and molecular mechanism of KRGE on the senescence of human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were grown in normal or KRGE-supplemented medium. Furthermore, they were transfected with heme oxygenase-1 (HO-1) gene or treated with its inhibitor, a NF-κB inhibitor, and a miR-155-5p mimic or inhibitor. Senescence-associated characteristics of endothelial cells were determined by biochemical and immunohistochemical analyses. Results: Treatment of HUVECs with KRGE resulted in delayed onset and progression of senescence-associated characteristics, such as increased lysosomal acidic β-galactosidase and decreased telomerase activity, angiogenic dysfunction, and abnormal cell morphology. KRGE preserved the levels of anti-senescent factors, such as eNOS-derived NO, MnSOD, and cyclins D and A: however, it decreased the levels of senescence-promoting factors, such as ROS, activated NF-κB, endothelial cell inflammation, and p21 expression. The beneficial effects of KRGE were due to the induction of HO-1 and the inhibition of NF-κB-dependent biogenesis of miR-155-5p that led to the downregulation of eNOS. Moreover, treatment with inhibitors of HO-1, NF-κB, and miR-155-5p abolished the anti-senescence effects of KRGE. Conclusion: KRGE delayed or prevented HUVEC senescence through a signaling cascade involving the induction of HO-1, the inhibition of NF-κB-dependent miR-155-5p biogenesis, and the maintenance of the eNOS/NO axis activity, suggesting that it may protect against vascular diseases associated with endothelial senescence.

Upregulation of heme oxygenase-1 by ginsenoside Ro attenuates lipopolysaccharide-induced inflammation in macrophage cells

Sokho Kim,Myung-Hoon Oh,Bum-Seok Kim,Won-Il Kim,Ho-Seong Cho,Byoung-Yong Park,Chul Park,Gee-Wook Shin,Jungkee Kwon 고려인삼학회 2015 Journal of Ginseng Research Vol.39 No.4

Background: The beneficial effects of ginsenoside species have been well demonstrated in a number of studies. However, the function of ginsenoside Ro (GRo), an oleanane-type saponin, has not been suffi- ciently investigated. Thus, the aim of the present study was to investigate the anti-inflammatory effects of GRo in vitro using the Raw 264.7 mouse macrophage cell line treated with lipopolysaccharide (LPS), and to clarify the possible mechanism of GRo involving heme oxygenase-1 (HO-1), which itself plays a critical role in self-defense in the presence of inflammatory stress. Methods: Raw 264.7 cells were pretreated with GRo (up to 200mM) for 1 h before treatment with 1 mg/ mL LPS, and both cell viability and inflammatory markers involving HO-1 were evaluated. Results: GRo significantly increased cell viability in a dose dependent manner following treatment with LPS, and decreased levels of reactive oxygen species and nitric oxide. GRo decreased inflammatory cytokines such as nitric oxide synthase and cyclooxygenase-2 induced by LPS. Moreover, GRo increased the expression of HO-1 in a dose dependent manner. Cotreatment of GRo with tin protoporphyrin IX, a selective inhibitor of HO-1, not only inhibited upregulation of HO-1 induced by GRo, but also reversed the anti-inflammatory effect of GRo in LPS treated Raw 264.7 cells. Conclusion: GRo induces anti-inflammatory effects following treatment with LPS via upregulation of HO-1.

Carbon monoxide prevents TNF-α-induced eNOS downregulation by inhibiting NF-κB-responsive miR-155-5p biogenesis

Choi, Seunghwan,Kim, Joohwan,Kim, Ji-Hee,Lee, Dong-Keon,Park, Wonjin,Park, Minsik,Kim, Suji,Hwang, Jong Yun,Won, Moo-Ho,Choi, Yoon Kyung,Ryoo, Sungwoo,Ha, Kwon-Soo,Kwon, Young-Guen,Kim, Young-Myeong Nature Publishing Group 2017 Experimental and molecular medicine Vol.49 No.11

<P>Heme oxygenase-1-derived carbon monoxide prevents inflammatory vascular disorders. To date, there is no clear evidence that HO-1/CO prevents endothelial dysfunction associated with the downregulation of endothelial NO synthesis in human endothelial cells stimulated with TNF-α. Here, we found that the CO-releasing compound CORM-2 prevented TNF-α-mediated decreases in eNOS expression and NO/cGMP production, without affecting eNOS promoter activity, by maintaining the functional activity of the <I>eNOS</I> mRNA 3′-untranslated region. By contrast, CORM-2 inhibited MIR155HG expression and miR-155-5p biogenesis in TNF-α-stimulated endothelial cells, resulting in recovery of the 3′-UTR activity of <I>eNOS</I> mRNA, a target of miR-155-5p. The beneficial effect of CORM-2 was blocked by an NF-κB inhibitor, a miR-155-5p mimic, a HO-1 inhibitor and siRNA against HO-1, indicating that CO rescues TNF-α-induced eNOS downregulation through NF-κB-responsive miR-155-5p expression via HO-1 induction; similar protective effects of ectopic HO-1 expression and bilirubin were observed in endothelial cells treated with TNF-α. Moreover, heme degradation products, except iron and <I>N</I>-acetylcysteine prevented H<SUB>2</SUB>O<SUB>2</SUB>-mediated miR-155-5p biogenesis and eNOS downregulation. These data demonstrate that CO prevents TNF-α-mediated eNOS downregulation by inhibiting redox-sensitive miR-155-5p biogenesis through a positive forward circuit between CO and HO-1 induction. This circuit may play an important preventive role in inflammatory endothelial dysfunction associated with human vascular diseases.</P>

GIS기반 동남 광역권 산업체 정보시스템 구축 및 활용

남광우 ( Kwang Woo Nam ),권일화 ( Il Hwa Kwon ),박준호 ( Jun Ho Park ) 한국지리정보학회 2014 한국지리정보학회지 Vol.17 No.1

광역 교통망 및 통신기술의 발전에 따라 광역경제권의 활성화를 위한 협력체계의 중요성이 날로 증대되고 있다. 이에 본 연구는 동남권 광역경제권을 대상으로 GIS 기반의 산업체 정보 공유를 목적으로 동남권 산업체 정보시스템을 구축하였다. 동남권의 경우 제조업 중심의 산업 집적지로서 행정경계를 넘어 산업체간의 근접성과 연결성을 고려한 효율적인 산업 클러스터 및 협력체계 운영이 요구된다. 정보시스템의 DB 구축을 위해 부산, 울산, 경남에서 구축되어진 산업체 DB를 활용하여 정보시스템을 구축한 바, 각 지자체간 데이터의 불일치와 GIS기반의 위치정보 미흡으로 인한 여러 가지 문제점을 발견하였다. 이의 해결을 위해서는 수집 및 배포, 활용과정을 고려한 표준화가 시급한 것으로 분석되었다. 본 연구는 산업체 정보의 공간정보화를 위해 지적도와 수치지도를 활용하여 웹상에서 관리자-사용자 쌍방향간의 정보생성 및 단계별 접근이 가능한 2-way 방식의 산업체 정보시스템을 구축하였다. 본 연구의 결과는 동남권 산업육성을 위한 협력체계 및 공동대응을 위한 산업체 정보 공유체계의 기초적 틀을 제공하였다는 데 그 의의가 있다고 하겠다. Following the technology developments of traffic network and communication for the wide area, the importance of the cooperation system to vitalize the wide area economy is increasing. Therefore, in this study, DongNam Kwon industry information system is established for the industrial information sharing based on GIS in the DongNam Kwon wide area economy. The DongNam Kwon is an industrial integration area centered with the manufacturing so that the operation of effective industrial cluster and cooperation systems are required across the administrational boundaries. To establish the database of the information, the information system was established utilizing already established industrial databases in Busan, Ulsan and Gyeongnam. But, various issues caused by the discordances among the data of each local government and the insufficiency of GIS based location information have been found. According to the analysis, the standardization considering the courses of collection, distributions and utilization are required immediately to solve the issues. This study establishes an 2-way industrial information system enabling the information creation and the phased approach between the administrator and the user in the bi-directions on the web by utilizing cadastral and numerical maps. The result of this study would have a meaning in providing a fundamental frame for cooperative responses and cooperation system for DongNam Kwon``s industrial promotion using industrial information sharing.

소규모 하수처리를 위한 파일럿 규모 이중슬러지 KNR^® (Kwon's nutrient removal) 시스템의 영얌염류 제거성능 평가

안진영,권중천,김윤학,정유훈,김두언,유선호,김병우,An, Jin-Young,Kwon, Joong-Chun,Kim, Yun-Hak,Jeng, Yoo-Hoon,Kim, Doo-Eon,Ryu, Sun-Ho,Kim, Byung-Woo 한국청정기술학회 2006 청정기술 Vol.12 No.2

본 연구에서 소규모 하수고도처리를 위한 이중슬러지(Dual sludge) $KNR^{(R)}$ (Kwon's nutrient removal) 시스템이 개발되었다. $KNR^{(R)}$ 시스템은 부유성장식 탈질미생물과 부착성장식 질산화미생물을 분리시킨 이중슬러지 공정으로 최초침전조, 혐기조, 무산소조, 농축조의 복합기능을 수행하는 UMBR (Upflow multi-layer bioreactor)과 펠렛형 담체가 충진된 호기성 담체조로 구성되어 있다. 소규모 하수처리시 본 개발공정의 안정성과 처리성능을 평가하기 위해 처리용량 $50m^3/d$ 규모의 파일럿 플랜트를 고도처리 공정으로 개선공사 중인 처리용량 $50m^3/d$ 규모의 실제 소규모 마을하수처리장에 적용하였다. UMBR과 담체조의 체류시간은 각각 4.7 h와 7.2h이었으며, 반응조 수온은 $18.1{\sim}28.1^{\circ}C$이었다. 유입 하수량과 유입수의 BOD/N의 변동폭이 컸음에도 불구하고 파일럿 플랜트는 안정된 처리성능을 보였다. 전체 실험기간 중 처리수의 $COD_{cr}$, $COD_{Mn}$, $BOD_5$, TN, TP의 평균 농도는 11.0 mg/L, 8.8 mg/L, 4.2 mg/L, 3.5 mg/L, 9.8 mg/L, 0.87/0.17 mg/L (poly aluminium chloride(PAC) 투입/미투입)이었으며, 제거율을 각각 95.3%, 87.6%, 96.3%, 96.5%, 68.2%, 55.4/90.3% 이었다. 잉여슬러지 발생량은 $A_2O$와 Bardenpho 등과 같은 단일슬러지를 이용하는 고도처리공정과 비교시 약 1.9~3.8배 낮은 $0.026kg-DS/m^3$ and 0.220 kg-DS/kg-BOD로 나타났다. A simple dual sludge process, called as $KNR^{(R)}$ (Kwon's Nutrient Removal) system, was developed for small sewage treatment. It is a hybrid system that consists of an UMBR (Upflow multi-layer bioreactor) as anaerobic and anoxic reactor with suspended denitrifier and a post aerobic biofilm reactor, filled with pellet-like media, with attached nitrifier. To evaluate the stability and performance of this system for small sewage treatment, the pilot-scale $KNR^{(R)}$ plant with a treatment capacity of $50m^3/d$ was practically applied to the actual sewage treatment plant, which was under retrofit construction during pilot plant operation, with a capacity of $50m^3/d$ in a small rural community. The HRTs of a UMBR and a post aerobic biofilm reactor were about 4.7 h and 7.2 h, respectively. The temperature in the reactor varied from $18.1^{\circ}C$ to $28.1^{\circ}C$. The pilot plant showed stable performance even though the pilot plant had been the severe fluctuation of influent flow rate and BOD/N ratio. During a whole period of this study, average concentrations of $COD_{cr}$, $COD_{Mn}$, $BOD_5$, TN, and TP in the final effluent obtained from this system were 11.0 mg/L, 8.8 mg/L, 4.2 mg/L, 3.5 mg/L, 9.8 mg/L, and 0.87/0.17 mg/L (with/without poly aluminium chloride(PAC)), which corresponded to a removal efficiency of 95.3%, 87.6%, 96.3%, 96.5%, 68.2%, and 55.4/90.3%, respectively. Excess sludge production rates were $0.026kg-DS/m^3$-sewage and 0.220 kg-DS/kg-BOD lower 1.9 to 3.8 times than those in activated sludge based system such as $A_2O$ and Bardenpho.

Synthesis of Sr Doped HoCrO3 Solid Solution Systems and Their Electrical Properties

PARK, SUNG HO,Kwon, Tae Yun 한국공업화학회 2000 Journal of Industrial and Engineering Chemistry Vol.6 No.4

Ho_(1-x)SrxCrO₃ systems, with perovskite structures, were synthesized for x=0, 0.05, 0.1, 0.2 at 1300 ℃ under atmospheric pressure. The results of X-ray analysis reveals that these prepared systems forms orthorhombic structures. Also, the HoCrO₃ systems which were doped with Sr cations on the site of Ho cations, exhibited a linear increase in their cell volume. It was found that the lattice parameter and reduced volume increased as the doping level of Sr^(2+) increased. From the measurements of D.C conductivity, the activation energy obtained from the conductivity dependence on temperature was 0.18 eV. The conduction mechanism would appear to be the hopping model of electron conduction.

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>