Mycophenolic Acid가 PDGF에 의해 증식 유도된 흰쥐 혈관평활근 세포에서 NAD(P)H Oxidase, 세포내 활성산소족 및 Mitogen-activated Protein Kinases에 미치는 영향

박제현,하헌주,김명수,허규하,김유선 이화여자대학교 약학연구소 2004 藥學硏究論文集 Vol.- No.14

Background: Vascular smooth muscle cell (VSMC) proliferation plays an important role in the development and progression of chronic allograft vasculopathy as in atherosclerosis. We already reported that mycophenolic acid (MPA) inhibited VSMC proliferation, cellular reactive oxygen species (ROS) and mitogen-activated protein kinases (MAPK) in human VSMCs. In this study, we examined further molecular mechanisms involved in the anti-proliferative effect of MPA in rat VSMCs. Methods: Primary rat VSMCs were stimulated with PDGF-BB 10 ng/mL in the presence or absence of MPA and various kinds of cell signaling inhibitors. Cell proliferation was assessed by [H³]-thymidine incorporation, NAD(P)H oxidase subunits mRNA expression by RT-PCR, dichlorofluorescein-sensitive cellular ROS by FACS, and the activation of PDGF receptor-β (Tyr 751), racl, and MAPK by Western blot analysis. Results: PDGF increased cell proliferation and cellular ROS, activation of PDGF receptor-β (Tyr 751), racl, expression of p22phox and MOX1 mRNA, ERK 1/2, and p38 MAPK, compared to control. MPA inhibited up-regulation of racl phosphorylation, p22phox and MOX1 mRNA expression, cellular ROS, and phosphorylation of ERK 1/2 and p38 MAPK. However, MPA did not affect PDGF receptor-β (Tyr 751) activation. Wortmannin, diphenyleniodonium (DPI), trolox, and NAC, each inhibited PDGF-induced ERK 1/2 and p38 MAPK activation. PD98059 and p38 MAPK inhibitor also inhibited PDGF-induced cell proliferation. Conclusion: These results suggest that MPA inhibits PDGF-induced VSMC proliferation through inhibiting NAD(P)H oxidase-dependent cellular ROS leading to ERK 1/2 and p38 MAPK activation. 배경 : 혈관평활근 세포의 증식은 장기이식 후 발생하는 혈관경화증이나 동맥경화증의 발생과 진행에 중요한 역할을 한다. Mycophenolic acid (MPA)는 강력한 면역억제제로서 혈관평활근 세포의 증식도 억제한다. 본 연구는 사람의 혈관평활근 세포에서 MPK가 세포증식에 관여하는 신호전달계인 활성산소족과 mitogen-activated protein kinases(MAPK) 활성화를 억제하는 기존의 연구결과를 바탕으로, PDGF에 의해 증식 유도된 흰쥐 혈관평활근 세포에서 MPA가 세포의 증식을 억제하는 기전을 검색하였다. 방법 : 일차 배양한 흰쥐 혈관평활근 세포를 PDGF-BB 10 ng/mL로 자극하였고, MPA를 비롯한 여러 종류의 신호전달 억제제는 PDGF를 투여하기 1시간 전에 투여하였다. 세포증식은 [H³]-thymidine incorporation으로, NAD(P)H oxidase subunit의 mRNA 표현은 RT-PCR로, dichlorofluorescein에 민감한 세포내 활성산소족은 FACS 방법으로, 그리고 PDGF 수용체-β (Tyr 751), racl 및 MARK 활성하는 Western blot으로 각각 분석하였다. 결과 : PDGF는 PDGF 수용체-β (Tyr 751)의 활성화 및 NAD(P)H oxidase subunit 중 racl의 활성화와 p22phox와 MOX1의 mRNA 표현을 대조군에 비하여 유의하게 증가시켰다. MPA는 PDGF 수용체의 활성화에는 영향을 주지 않았으나, racl의 활성화, p22phox와 MOX1의 mRNA 표현의 상향 조절, 세포내 활성산소족, 그리고 ERK 1/2와 p38 MAPK의 활의한 EFK 1/2와 p38 MAPK 활성화와 세포 증식을 억제하였다. PD98059와 p38 MAPK 억제제는 PDGF에 의한 혈관평활근 세포의 증식을 억제하였다. 결론 : 본 연구결과는 MPA가 NAD(P)H oxidase를 억제함으로써 세포내 활성산소족 생산과, MAPK 활성화를 억제함으로써 PDGF에 의하여 유도되는 혈관평활근 세포의 증식을 억제함을 시사하였다.

Mycophenolic Acid와 Rapamycin이 흰쥐 사구체 혈관간세포증식과 세포외기질 생성에 미치는 영향

김명수,박제현,하헌주,허규하,서지연,김유선,김혜진,박기일 이화여자대학교 약학연구소 2004 藥學硏究論文集 Vol.- No.14

Background: Excess proliferation and extracellular matrix (ECM) accumulation of mesenchymal cells such as vascular smooth muscle cells (VSMC) and glomerular mesangial cells cause chronic allograft nephropathy showing transplant vascular sclerosis and glomerulosclerosis. Mycophenolic acid (MPA) and rapamycin (RPM) are well known as strong inhibitors of VSMC proliferation, but their effects on the glomerular mesangial cells are not yet clearly understood. This study examined the effects of MPA or RPM on PDGF-induced proliferation and ECM accumulation in rat glomerular mesangial cells. Methods: Mesangial cells isolated from the glomeruli of Sprague-Dawley rats were cultured with DMEM containing 20% fetal bovine serum. Growth arrested and synchronized cells were administered with test drugs (MPA10 nM-10μM, RPM 0.1 nM-1μM) before the addition of PDGF 10 ng/mL. Cell proliferation was assessed by [³H]thymidine incorporation, collagen by [³H]proline incorporation, and fibronectin, ERK, and p38 MAPK by Western blot analysis. Results: PDGF increased mesangial cell proliferation by 4.64-fold. Compared to stimulated control, MPA above 500 nM and RPM above 10nM showed a significant inhibitory effect in a dose-dependent manner. The IC_(50) of MPA and RPM against PDGF-induced mesangial cell proliferation were around 500 nM and 100 nM, respectively. The collagen synthesis was also inhibited by MPA and RPM, but the fibronectin secretion was inhibited by MPA alone. The proliferation of mesangial cell correlated with activation of ERK and. p38 MAPK. MPA, but not RPM, inhibited ERK and p38 MAPK activation. Conclusion: This study demonstrated that MPA and RPM significantly inhibited PDGF-induced proliferation and ECM production in rat glomerular mesangial cells. The inhibitory effects of MPA, but not RPM, are correlated with ERK and p38 MAPK.

ApoB/ApoA-I ratio is independently associated with carotid atherosclerosis in type 2 diabetes mellitus with well-controlled LDL cholesterol levels

( Ji Eun Jun ),( Young Ju Choi ),( Yong-ho Lee ),( Dae Jung Kim ),( Seok Won Park ),( Byung Wook Huh ),( Eun Jig Lee ),( Sun-ha Jee ),( Kyu Yeon Hur ),( Sung Hee Choi ),( Kap Bum Huh ) 대한내과학회 2018 The Korean Journal of Internal Medicine Vol.33 No.1

Background/Aims: This study aimed to investigate whether the apolipoprotein (Apo) B/ApoA-I ratio is associated with carotid intima-media thickness (CIMT) in type 2 diabetes mellitus (T2DM) subjects with low density lipoprotein cholesterol (LDL-C) levels less than 100 mg/dL. Methods: This cross-sectional study included 845 subjects aged with T2DM 40 to 75 years who had visited Huh’s Diabetes Center in Seoul, Republic of Korea for CIMT measurement. Traditional fasting lipid profiles, ApoB and ApoA-I levels were examined. CIMT was measured at three points on the far wall of 1 cm long section of the common carotid artery in the proximity of the carotid bulb. The mean value of six measurements from right and left carotid arteries were used as the mean CIMT. In this study, carotid atherosclerosis was defined as having a focal plaque or diffuse thickening of the carotid wall (mean CIMT ≥ 1.0 mm) Results: The prevalence of carotid atherosclerosis increased with ApoB/ApoA-I ratio. The ApoB/ApoA-I ratio, expressed as both quartiles (odds ratio [OR], 2.14; 95% confidence interval [CI], 1.21 to 3.79; p for trend = 0.014) and continuous values (OR, 10.05; 95% CI, 3.26 to 30.97; p < 0.001), was significantly associated with a higher risk for carotid atherosclerosis, regardless of conventional cardiovascular disease risk factors. The optimal ApoB/ApoA-I ratio cutoff value for detecting carotid atherosclerosis was 0.57, based on receiver operating characteristic curve analysis with a sensitivity of 58.0% and a specificity of 55.1%. Conclusions: A high ApoB/ApoA-I ratio was significantly associated with carotid atherosclerosis in T2DM patients with LDL-C levels less than 100 mg/dL.

Chronologically Different Impact of Immunologic and Non-Immunologic Risk Factors on Renal Allograft

Kyu Ha Huh,Myoung Soo Kim,Soon Il Kim,Jong Hoon Lee,Kiil Park,Yu Seun Kim 대한외과학회 2004 대한외과학회 학술대회 초록집 Vol.2004 No.10

항문암: 종류, 치료 및 예후에 대한 고찰

허규하,이강영,하현수,박재균,김남규,손승국,민진식 대한대장항문학회 2002 Annals of Coloproctolgy Vol.18 No.5

Identification and molecular characterization of PERV Gamma1 long terminal repeats

Huh, Jae-Won,Kim, Dae-Soo,Ha, Hong-Seok,Ahn, Kung,Chang, Kyu-Tae,Cho, Byung-Wook,Kim, Heui-Soo Korean Society for Molecular Biology 2009 Molecules and cells Vol.27 No.1

Porcine endogenous retroviruses (PERVs) gamma1 in the pig genome have the potential to act as harmful factors in xenotransplantation (pig-to-human). Long terminal repeats (LTRs) are known to be strong promoter elements that could control the transcription activity of PERV elements and the adjacent functional genes. To investigate the transcribed PERV gamma1 LTR elements in pig tissues, bioin-formatic and experimental approaches were conducted. Using RT-PCR amplification and sequencing approaches, 69 different transcribed LTR elements were identified. And 69 LTR elements could be divided into six groups (15 subgroups) by internal variation including tandem repeated sequences, insertion and deletion (INDEL). Remarkably, all internal variations were indentified in U3 region of LTR elements. Taken together, the identification and characterization of various PERV LTR transcripts allow us to extend our knowledge of PERV and its transcriptional study.

Effect of non-Newtonian viscosity on the fluid-dynamic characteristics in stenotic vessels

Huh, Hyung Kyu,Ha, Hojin,Lee, Sang Joon Springer-Verlag 2015 Experiments in fluids Vol.56 No.8

Carbon Fiber as Material for Radiation Fixation Device : A comparative study with acrylic

Eui Kyu Chie,Jang Pil Park,Soon Nyung Huh,Semie Hong,In-Ah Kim,Hong-Gyun Wu,Jae-Sung Kim,Wee-Saing Kang,Il Han Kim,Sung Whan Ha,Charn Il Park 대한방사선방어학회 2005 방사선방어학회지 Vol.30 No.1

Altered ER–mitochondria contact impacts mitochondria calcium homeostasis and contributes to neurodegeneration in vivo in disease models

Lee, Kyu-Sun,Huh, Sungun,Lee, Seongsoo,Wu, Zhihao,Kim, Ae-Kyeong,Kang, Ha-Young,Lu, Bingwei National Academy of Sciences 2018 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF Vol.115 No.38

<P><B>Significance</B></P><P>Ca<SUP>2</SUP><SUP>+</SUP> regulates cellular metabolism, proliferation, and differentiation. Ca<SUP>2+</SUP> homeostasis is critical for cellular function and health. Mitochondria help buffer transient Ca<SUP>2+</SUP> elevations and prevent cell death induced by Ca<SUP>2+</SUP> overload. Mito-Ca<SUP>2+</SUP> is also required for optimal activity of certain key mitochondrial functions, such as oxidative phosphorylation and metabolism. Thus, mito-Ca<SUP>2+</SUP> homeostasis assumes central roles in cellular health. Endoplasmic reticulum (ER) and mitochondria make intimate contacts and exchange molecules such as Ca<SUP>2+</SUP> and lipids. We find that ER-to-mitochondria Ca<SUP>2+</SUP> transfer is important for mito-Ca<SUP>2+</SUP> homeostasis and that the conserved Miro protein is critically involved. We show that mito-Ca<SUP>2+</SUP> homeostasis is disrupted in neurodegenerative disease models and its restoration is beneficial. Our findings have important implications for therapeutic intervention of neurodegenerative diseases.</P><P>Calcium (Ca<SUP>2+</SUP>) homeostasis is essential for neuronal function and survival. Altered Ca<SUP>2+</SUP> homeostasis has been consistently observed in neurological diseases. How Ca<SUP>2+</SUP> homeostasis is achieved in various cellular compartments of disease-relevant cell types is not well understood. Here we show in <I>Drosophila</I> Parkinson’s disease (PD) models that Ca<SUP>2+</SUP> transport from the endoplasmic reticulum (ER) to mitochondria through the ER–mitochondria contact site (ERMCS) critically regulates mitochondrial Ca<SUP>2+</SUP> (mito-Ca<SUP>2+</SUP>) homeostasis in dopaminergic (DA) neurons, and that the PD-associated PINK1 protein modulates this process. In <I>PINK1</I> mutant DA neurons, the ERMCS is strengthened and mito-Ca<SUP>2+</SUP> level is elevated, resulting in mitochondrial enlargement and neuronal death. Miro, a well-characterized component of the mitochondrial trafficking machinery, mediates the effects of PINK1 on mito-Ca<SUP>2+</SUP> and mitochondrial morphology, apparently in a transport-independent manner. Miro overexpression mimics <I>PINK1</I> loss-of-function effect, whereas inhibition of Miro or components of the ERMCS, or pharmacological modulation of ERMCS function, rescued <I>PINK1</I> mutant phenotypes. Mito-Ca<SUP>2+</SUP> homeostasis is also altered in the LRRK2-G2019S model of PD and the PAR-1/MARK model of neurodegeneration, and genetic or pharmacological restoration of mito-Ca<SUP>2+</SUP> level is beneficial in these models. Our results highlight the importance of mito-Ca<SUP>2+</SUP> homeostasis maintained by Miro and the ERMCS to mitochondrial physiology and neuronal integrity. Targeting this mito-Ca<SUP>2+</SUP> homeostasis pathway holds promise for a therapeutic strategy for neurodegenerative diseases.</P>

Dynamic Evolution of tRNA(Thr)-Derived HpaI SINEs and Effect on Genomes of Oncorhynchus Species

Jae Won Huh,Dae Soo Kim,Yu Na Noh,Sang Je Park,Hong Seok Ha,Choong Gon Kim,Youn Ho Lee,Chang Keun Kang,Kyu Tae Chang,Heui Soo Kim 한국유전학회 2009 Genes & Genomics Vol.31 No.3

Short interspersed nuclear elements (SINEs) are the most abundant non-autonomous retroelements in many vertebrate genomes. The events that led to their integration may have had marked effects on the evolution of host genomes. One well-investigated SINE lineage is in the pacific salmon (genus Oncorhynchus). Experimental approaches and bioinformatics have been used to investigate the dynamic features and evolutionary impact of these SINEs. Four gene-related HpaI SINEs in the CD4L-2a, NOS, MHC and IL1B genes were identified by bioinformatics tool. To investigate these SINEs, PCR amplification and sequencing were performed on eight species of the genus Oncorhynchus and one of Salmo. Unexpectedly, the CD4L-2a, MHC and IL-1B gene loci proved to be dimorphic for the HpaI SINE insertion; this may be attributable to lineage sorting. Sequence transduction and horizontal transmission events also occurred in CD4L-2a. To elucidate the impact of HpaI SINEs on pacific salmon genomes and the diversity of transcriptomes, 243,668 mRNA sequences from the GenBank database were analyzed. A total of 163 mRNA sequences were identified as fused with HpaI SINEs. Among these, 87 ESTs were annotated into 41 functional genes. Our data suggest that SINEs could contribute to the genomic diversity of the pacific salmon by exonization and could move more dynamically within this genome by lineage sorting, sequence transduction and horizontal transmission.