Combined Gene Therapy with Hypoxia-Inducible Factor-1α and Heme Oxygenase-1 for Therapeutic Angiogenesis

Bhang, Suk Ho,Kim, Ju Hee,Yang, Hee Seok,La, Wan-Geun,Lee, Tae-Jin,Kim, Ga Hee,Kim, Hyun Ah,Lee, Minhyung,Kim, Byung-Soo Mary Ann Liebert 2011 Tissue engineering. Part A Vol.17 No.7

<P>Transfection with either hypoxia-inducible factor-1α (HIF-1α) or heme oxygenase-1 (HO-1) gene can induce neovascularization in ischemic tissues. Although expression of transfected HIF-1α gene occurs rapidly, the expressed HIF-1α protein degrades quickly, limiting its therapeutic efficacy. Meanwhile, expressed HO-1 protein does not rapidly undergo degradation, but gene expression occurs a couple of days after transfection, resulting in apoptosis and a delay in angiogenesis in ischemic tissues at the incipient period of HO-1 gene transfection. We hypothesize that combined delivery of HIF-1α and HO-1 gene will enhance antiapoptosis and neovascularization in ischemic tissue compared with HIF-1α or HO-1 single-gene therapy. To test this hypothesis, ischemic mouse hindlimbs were treated with HIF-1α and/or HO-1 gene therapy. The combined gene therapy proved superior to both single-gene therapies, resulting in rapid expression of HIF-1α gene and long-term maintenance of expressed HO-1 protein. The apoptosis in the ischemic region was significantly less, and angiogenic growth factor secretion and angiogenesis were greater in the combined gene therapy than in either of the single-gene therapies. Our results suggest that a combined gene therapy of HIF-1α and HO-1 enhances the transfection of both genes and improves angiogenesis compared with either single-gene therapy.</P>

운동강도의 차이가 대장에서의 Heme oxygenase-1 발현에 미치는 영향: Oligonucleotide chip microarray analysis

최은주 ( Eun Ju Choi ),류호영 ( Ho Young Ryu ),차광석 ( Kwang Suk Cha ) 한국운동생리학회(구-한국운동과학회) 2012 운동과학 Vol.21 No.1

이 연구의 목적은 운동강도 차이에 의한 대장조직이 Heme oxygenase-1(HO-1)의 mRNA 발현에 어떠한 영향을 미치는지 규명하기 위해, Sprage-Dawley계 흰쥐를 대상으로 통제그룹(CON)과 저강도 운동그룹(LIE), 고강도 운동그룹(HIE)으로 구분하여 4주 동안 트레드밀운동 후 48시간 이후에 High through-put microarray analysis방법으로 다음과 같은 실험을 실시하였다. 본 실험은 Rat ABI oligo chip 26,857 개의 유전자 중 filtering을 통하여 12.079개 유전지를 선택하였고, 이 중 유의성 있는(p<.05) 유전자 12,072개를 선별하였으며, clustering분석 방법과 면역조직화학염색법에서도 HO-l이 운동강도에 따라 유의하게 발현하였다(p<.05). Microarray분석 후 RT-PCR로 확인한 결과 HO-I의 유전자 발현양상이 일치하는 결과를 얻었다. 이 실험의 결과로서 운동이 대장에서 HO-1 발현에 영향을 주는 것을 알 수 있었다. 운동이 대장에서의 mRNA 발현이 저강도 운동그룹(LIE)보다 고강도 운동그룹(HIE)에서 더 많이 발현되어지는 것으로 보아 고강도 운동에 의해 발생되는 산소라디칼을 HO-l의 유도를 통하여 적절히 제거함으로써 스스로 항상성을 유지하고 있는 것으로 생각되어진다. The purpose of this study is to identify how large intestine tissue effects to mRNA expression in Heme oxygenase-l (HO-1) due to differences in exercise intensity. To perform this experiment we divided Sprage-Dawley rats into 3 groups (control group (CON). low-intensity exercise group (LIE) and high-intensity exercise group (HIE)), and in 48 hours after 4 weeks treadmill exercise. the following experiment was performed with "High through-put micro array analysis methods" . 12,079 genes were chosen by filtering of the Rat ASI oligo chip 26.857 genes in this experiment Among 12.079 genes, we selected significant genes (p<.05) that were also expressed in the ways of clustering analysis and immunohistochemistry. With results of RT-PCR confirming and microarray analyzing, we derived that gene expression profiles of HO-l had been consistent. As a result of this experiment, we certainly identified that exercise effects to HQ-l expression in large intestine. mRNA expression was more expressed in high-intensity exercise group CHIE) than low-intensity exercise group (LIE), it seems that homeostasis maintains itself by properly removing oxygen radical. caused by high intensity exercise. through reduction of HO-l.

Adenovirus-mediated heme oxygenase-1 gene transfer to neonatal porcine islet-like cluster cells: the effects on gene expression and protection from cell stress

염혜정,Han Ro,Sol Ji Park,Ju Ho Hong,Bumrae Cho,김화정,Sung Joo Kim,황종익,이병천,안규리,양재석 한국바이오칩학회 2012 BioChip Journal Vol.6 No.1

Porcine islet xenotransplantation is a promising strategy for the treatment of diabetes that overcomes donor shortages. However, islet xenografts are susceptible to oxidative stress and apoptosis. Heme oxygenase-1 (HO-1) has been shown to protect cells from oxidative stress, apoptosis and inflammation. Here, we investigated whether introduction of human HO-1 (hHO-1) into neonatal porcine islet-like cell clusters (NPCCs) can induce beneficial transcriptional changes in NPCCs against cellular stress. NPCCs were transduced with either adenovirus-HO-1 (Ad-HO-1) or control adenovirus-GFP (Ad-GFP). After treatment with hydrogen peroxide (H2O2) for 24 hours, nitrite oxide (NO) production assays were performed to detect oxidative stress. Microarray analysis was performed using a pig oligonucleotide 44 K gene chip. We profiled transcriptional changes to apoptosis, oxidant and inflammatory genes, and real-time PCR analysis was also performed to confirm the microarray results. Survival of NPCCs after treatment with H2O2 was significantly higher in the Ad-HO-1 group (p⁄0.001), and NO production also decreased in the Ad-HO-1 group (p⁄0.01). The microarray results showed that the expression of pro-apoptosis genes such as CASP3, CASP7, CASP10, CIDE-B and CIDE-C was significantly decreased in the Ad-HO-1 virus group (CASP10; p⁄0.05, CASP3, CIDE-C; p⁄0.01, CASP7, CIDE-B; p⁄0.001). We also found that the expression of oxidative stresses genes including COX1, COX2, CYB5A, SDHD and NOS2 was decreased, and that the anti-oxidant genes Gpx1 and SOD2 were increased in the Ad-HO-1 group (NOS2; p⁄0.05, COXI, COX2, CYB5A, SDHD, SOD2, GPX1; p⁄0.001). However, inflammatory gene expression was not significantly changed. Realtime PCR analysis confirmed the results of the microarray analysis. These results shed light on the underlying mechanisms of the protective effects of hHO-1 on porcine islets from cellular stresses and suggest that hHO-1 could be a promising target gene for the production of transgenic pigs that confer improved islet xenograft survival.

Curcumin protects retinal pigment epithelial cells against oxidative stress via induction of heme oxygenase-1 expression and reduction of reactive oxygen

Woo, Je Moon,Shin, Da-Yong,Lee, Sung Ju,Joe, Yeonsoo,Zheng, Min,Yim, Jin Ho,Callaway, Zak,Chung, Hun Taeg Molecular Vision 2012 Molecular vision Vol.18 No.-

<P><B>Purpose</B></P><P>To determine whether curcumin induces expression of the defensive enzyme heme oxygenase-1 (HO-1) and protects cells against oxidative stress in cultured human retinal pigment epithelial cells.</P><P><B>Methods</B></P><P>Effective concentrations and toxicities of curcumin were determined after 3 h of curcumin treatment with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Confluent human retinal pigment epithelium cell lines (ARPE-19) were preincubated with curcumin and oxidatively challenged with H<SUB>2</SUB>O<SUB>2</SUB>. HO-1 expression was determined with western blot analysis. To confirm the protective role of HO-1 in oxidative stress, small interfering RNA (siRNA) against HO-1 or inhibitor of HO-1 was treated with curcumin in retinal pigment epithelium cells. Intracellular levels of reactive oxygen species (ROS) were measured with flow cytometry and confocal microscopy. Apoptosis was evaluated with Annexin V-fluoroscein isothiocyanate staining.</P><P><B>Results</B></P><P>Curcumin had little cytotoxicity at concentrations less than 30 μM, and HO-1 expression was the highest at the 15 μM concentration. At this concentration, curcumin also increased the cytoprotective effect against the oxidative stress of H<SUB>2</SUB>O<SUB>2</SUB> through the reduction of ROS levels in human retinal pigment epithelial cells. Curcumin’s effect on the reduction of ROS was mediated by the increase in HO-1 expression.</P><P><B>Conclusions</B></P><P>Curcumin upregulated the oxidative stress defense enzyme HO-1 and may protect human retinal pigment epithelial cells against oxidative stress by reducing ROS levels.</P>

Ho-166 부착풍선도자를 이용한 방사선 조사의 돼지 관상동맥 스텐트 재협착 예방 효과

김원 ( Kim Won ),정명호 ( Jeong Myeong Ho ),박옥영 ( Park Og Yeong ),정우곤 ( Jeong U Gon ),박우석 ( Park U Seog ),김주한 ( Kim Ju Han ),안영근 ( An Yeong Geun ),조정관 ( Jo Jeong Gwan ),박종춘 ( Park Jong Chun ),강정채 ( Kang Je 한국지질동맥경화학회 ( 구 한국지질학회 ) 2002 韓國脂質學會誌 Vol.12 No.1

배경 : 국내에서 개발된 방사선 동위원소 Holmium-166 (166Ho)은 주로 베타선을 방출하며, 166Ho을 부착한 풍선도자를 이용하여 돼지 관상동맥 재협착 모형에서 풍선확장술 후 신생내막 증식을 전신적 부작용 없이 안전하고 효과적으로 억제하였음을 보고한 바 있다. 본 연구에서는 돼지 관상동맥 스텐트 재협착 모형에서 스텐트 시술 후 신생내막 증식에 의한 재협착 병변을 166Ho 부착 풍선도자를 이용하여 치료하여 그 효과를 관찰하고자 하였다. 방법

간질환자의 간질 관련 외상

오동호,박운규,이영주,김희태,김승현,김주한,김명호,신동진 대한신경과학회 1998 대한신경과학회지 Vol.16 No.4

척수 경막외 혈관 기형에 의한 재발성 척수병증

오동호,김희태,박운규,이영주,김승현,김주한,김명호,박문향 대한신경과학회 1998 대한신경과학회지 Vol.16 No.5

황정(黃精) 추출물의 화학구조결정에 관한 연구 (Ⅰ)

윤중호,박주희,김정주,권기락,안철진,주우홍,강진호,신동수 Institute of Genetic Engineering Changwon National 1998 Gene and Protein Vol.2 No.1

본 연구에서는 황정 속에 포함되어 있는 생리활성 물질을 hexane, CHCl₃과 n-butanol층에서 각각의 성분들을 추출하였고, hexane층에서 분리된 화합물 I과 H 중에서, 화합물 I의 봐학구조를 ¹H-nmr, ??, DEPT135, COSY, HMQC, HMBC 스펙트럼 및 MS 스펙트럼 등의 분광학적인 방법에 의해 결정하였다. 화합물 I의 구조는 9,12-(9E, 12E)-octadecadienoic acid 임을 확인하였다. In this Paper, biologically active compounds were extracted using organic solvents as hexane. CHC1₃, n-butanol to give each component. Chemical structure of compound I was characterized using ¹H-nmr, ??, DEPT135, COSY, HMQC, HMBC spectrum and MS-spectrum, in separated compound I and Ⅱ from hexane layer. Finally, chemical structure of compound I was determined as 9,12-(9E,12E)-octadecadienoic acid.

공기 싸이클론 분리 장치의 분체 분리 효율

姜錫浩,朴永孝,金珠浩,朱星浩 嶺南大學校 工業技術硏究所 1985 연구보고 Vol.13 No.1

The collection efficiency of the Lapple's standard-type cyclone separator was determined in order to investigate the effects of the particle size of the dust samples, the dust loading, and the air inlet velocity to the cyclone under the ambient conditions. The experimental conditions are in the following ranges; the air inlet velocity; 5 to 14.7 m/s, the particle sizes of the mono-dispersed system; 14 to 280 ㎛, the dust loading; 17.8 to 58.6 g/㎥, respectively. The collection efficiencies determined were compared with those predicted by the semi-empirical equations proposed earlier by Strauss (³) and by Licht et al, (⁴) and those are shown to be increased, as the particle size of the sample dusts and the air inlet velocity increase, respectively. However, the effect of the dust loading in the air could not be recognized, but slightly increased. For the poly-dispersed particulate samples, having a wide particle size distribution, the grade or separation efficiency curve was experimentally determined to specify more exactly the cyclone separation performance, and the result was discussed with the statistical analysis previously worked by Herrmann.(??)

ORiginal Article : Gastroprotective Effects of PMK-S005 against Ethanol-Induced Acute Gastric Damage in Rats

( Yoon Jeong Choi ),( Nayoung Kim ),( Ju Yup Lee ),( Ryoung Hee Nam ),( Ji Hyung Seo ),( Seonmin Lee ),( Hee Jin Kim ),( Yoon Jin Choi ),( Hye Seung Lee ),( Dong Ho Lee ) 대한간학회 2016 Gut and Liver Vol.10 No.3

Background/Aims: This study aimed to examine the gastroprotective effects of PMK-S005, which is a synthetic S-allyl-Lcysteine (SAC; a sulfur-containing amino acid), against acute ethanol-induced gastric damage in rats. Methods: Sprague- Dawley rats were divided into six groups, including a nonethanol group, groups treated with absolute ethanol 1 hour after pretreatment with various doses of PMK-S005 (1, 5, and 10 mg/kg) or rebamipide (50 mg/kg), and an absolute ethanolonly group. Ethanol-induced gross ulcer and mucus levels were measured. Myeloperoxidase, tumor necrosis factor a, interleukin 1β, PGE2, LTB4, cPLA2, COX-1, and COX-2 levels were estimated by enzyme-linked immunosorbent assay or Western blot analysis. Furthermore, the protein expression levels of antioxidant enzymes, including heme oxygenase-1 (HO-1), NAD(P)H:quinine oxidoreductase 1 (NQO-1), GCLC, and GCLM, were assessed. Results: PMK-S005 significantly attenuated the ethanol-induced gastric damage; it reduced mucosal inflammatory cytokine production and increased mucus levels. The expression levels of cPLA2, COX-1, and COX-2 were decreased by PMK-S005. PMK-S005 did not affect PGE2 synthesis, but LTB4 production was significantly suppressed. In addition, long-term administration of PMKS005 significantly increased the expression of HO-1, NQO-1, GCLC, and GCLM. Conclusions: These results strongly suggest that PMK-S005 prevents gastric mucosal damage and that these gastroprotective activities are due to anti-inflammatory effects and enhancement of the gastric defense system, including antioxidant enzymes. (Gut Liver 2016;10:348- 355)