Uncoupling Protein 3의 골격근 세포내 과발현이 OLETF 백서 및 배양된 골격근 세포에서 포도당대사에 미치는 영향

한정희,박혜선,고정민,김하영,강호경,이인규,박중열,홍성관,이재담,이기업 대한당뇨병학회 2002 Diabetes and Metabolism Journal Vol.25 No.6

연구배경:Uncoupling protein(UCP)는 미토콘드리아의 내막에 위치하는 단백질로 세포내의 과다한 에너지를 열로 발산시키는 기능을 가진다. 최근 동물의 갈색지방조직에만 존재하는 UCP와 유사성을 가진 아형들(UCP2,3)이 사람에게도 존재함이 알려져 큰 관심을 끌도 있는데 이중 UCP3는 그 발현이 골격근세포와 갈색지방조직에만 국한된다. 본 연구에서는 UCP3가 체내 인슐린 감수성을 결정하는데 가장 중요한 조직인 골격근에 국한되어 발현되는 점에 착안하여 UCP3를 골격근세포에 과발현시켰을 때 포도당 대사에 어떠한 영향이 나타나는 지를 조사하였다. 방법:25주령의 8마리의 OLETF 백서를 대상으로 하여 4마리는 골격근에 adenovirus 2mL(1×10¹²pfu/mL)를 주사하여 대조군으로 하였고 4마리는 골격근에 재조합법으로 제작된 adenovirus­UCP3 2mL(1×10¹²pfu/mL)를 주사하였다(UCP3 과발현군). UCP3를 투여한 백서에서 먹이섭취가 증가하는 경향이 있어 그 전날 대조군이 먹은 야의 먹이만큼 투여하였다. 골격근에 adenovirus를 주사한 10일 후에 euglycemic hyperinsulinemic clamp를 시행하였다. Adenovirus­UCP를 C2C12 골격근 세포에 transfection시켜 UCP3를 C2C12 골격근 세포에 transfection시켜 UPS3­C2C12를 만들고 C2C12 골격근 세포와 UPS3­C2C12 골격근 세포에서 포도당 수송 및 당원합성을 측정하였다. 결과:UCP3 과발현 OLETF에서 체중이 감소하는 경향을 보였고 인슐린 감수성이 증가하였다. C2C12세포에서 기저상태 포도당 수송은 1.28±0.17μmol/L/min였고 100nM 인슐린으로 2시간 처리한 후 2.67±0.20 μmol/L/min로 증가하였다. UCP3­C2C12 세포에서는 기저상태 포도당 수송이 3.98±0.13μmol/L/min로 증가되었고 인슐린 처리 후 5.74±0.44μmol/L/min로 증가하였다. 인슐린을 처리한 UCP3­C2C12 세포에 P13K 억제제인 wortmannin을 첨가하였을 때 포도당 수송활성이 3.81±0.20μmol/L/min로 감소하였다. 기저상태 당원합성은 C2C12 세포에서 0.25±0.01μmol/L/min였고 인슐린 처리 후 0.45±0.01μmol/L/min로 증가하였다. UCP3­C2C12 세포에서는 기저상태 당원합성이 0.62±0.01μmol/L/min였고 인슐린 처리 후 1.26±454μmol/L/min로 증가하였다. UCP3­C2C12세포에 wortmannin을 첨가하였을 때 당원합성율이 0.80±0.04μmol/L/min로 감소하였다. 결론:UCP3 과발현이 OLETF 백서에서 인슐린 감수성을 증가시켰고 골격근세포에서 포도당 수송 및 당원합성을 증가시켰다. wortmannin을 첨가하였을 때 포도당 수송 및 당원합성이 감소함으로 보아 이 과정이 인슐린 신호전달체계인 P13K에 일부 의존함을 알 수 있었다. Background : UC P3 is a mitochondrial membrane protein expressed selectively in the skeletal muscle and brown adipose tissue. Since the skeletal muscle is the main organ determining insulin sensitivity in the body, it was hypothesized that UCP3 overexpression in skeletal muscle cells would improve glucose metabolism. Methods : An adenovirus-UCP3 was produced by a recombinant DNA method. OLETF rats were divided into 2 groups. Four rats were injected with the adenovirus-UCP3 (UCP3 group) and others were injected with the adenovirus(control group) in the skeletal muscle. The UCP3 group was provided with the same quantity of food as that consumed by the control group on the previous day. Insulin sensitivity was evaluated by the euglycemic hyperinsulinemic clamp method. In a separate experiment, glucose transport and glycogen synthesis we evaluated in C2C212 cells transfected with ether an adenovirus or the adenovirus-UCP3. Results : The insulin sensitivity improved significantly and the body weight decreased in the UCP3 group. The glucose transport and glycogen synthesis were higher in the UCP3-C2C12 skeletal muscle cells at the basal state. After insulin treatment, glucose transport and glycogen synthesis were also higher in the UCP3-C2C12 cells but the increments were reduced after treatment with wortmannin, a PI3K inhibitor. Conclusion : Insulin sensitivity was higher in the UCP3-overexpressed OLETF rats in the in vivo study. UCP3 transfection also increased glucose transport and glycogen synthesis in the cultured skeletal muscle cells by a PI3K dependent mechanism(J Kor Diabetes Asso 25 :460~468, 2001).

Comprehensive analysis of time- and dose-dependent patterns of gene expression in a human mesenchymal stem cell line exposed to low-dose ionizing radiation.

Jin, Young-Woo,Na, Young-Ji,Lee, Young-Ju,An, Sungkwan,Lee, Jung Eun,Jung, Meeseon,Kim, Heesun,Nam, Seon Young,Kim, Cha Soon,Yang, Kwang Hee,Kim, Seung Up,Kim, Woo Kyung,Park, Woong-Yang,Yoo, Keun-You National Hellenic Research Foundation 2008 ONCOLOGY REPORTS Vol.19 No.1

<P>We focused on the transcriptional responses induced by low and very low doses of ionizing radiation with time effect. Regardless of their importance only a few limited studies have been done. Here we applied a large-scale gene transcript profile to elucidate the genes and biological pathways. Immortalized human mesenchymal stem cells were irradiated with 0.01, 0.05, 0.2 and 1 Gy of gamma radiation and total RNA was extracted from each cell line at 1, 4, 12 and 48 h after exposure. The essential transcriptional responses were identified according to dose and time. A total of 6,016 genes showed altered expression patterns at more than one time point or dose level among the investigated 10,800 genes. Genes that showed dose-dependent expression responses were involved in signal transduction, regulation of transcription, proteolysis, peptidolysis and metabolism. Those that showed time-dependent responses were divided into two distinct groups: the up-and-down group was associated with 'cellular defense mechanisms' such as apoptosis, cell adhesion, stress response and immune response and the down-and-up group with 'fundamental cellular processes' such as DNA replication, mitosis, RNA splicing, DNA repair and translation initiation. Genes showing both dose-and time-dependent responses exhibited a mixture of both features. A highly non-linear relationship between the IR dose and the transcriptional relative response was obtained from the dose-dependent group. The time-dependent group also exhibited a non-linear relationship as the complex effect group did. Some of the early-reactive-phase (1-4 h) genes showed a differential expression response to 0.01, 0.05 and 0.2 Gy but were unresponsive to 1 Gy. Some of the late-recovery-phase (12-48 h) genes showed a differential expression to 1 Gy but were relatively unresponsive to other doses. We further characterized the gene expression patterns that could be implicated in the molecular mechanism of the cellular responses to low and very low-dose irradiation.</P>

혈장 중 시메티딘의 분석조건 설정 및 시메티딘 제제의 생물학적 동등성

김희규,이정민,윤미경,최성업,이상길,최영욱 중앙대학교 약학연구소 2002 약학 논총 Vol.16 No.-

Cimetidine is a histamine H_2-receptor antagonist, used for the treatment of endoscopically or radiographically conformed duodenal ulcer, pathologic GI hypersecretory condition, and active, benign, gastric ulcer. Simple method for determining of cimetidine in human plasma has been developed and validated. The procedure was linear in the range from 0.05 to 2 ug/mL for cimetidine. The intraday and interday validation for coefficient of variance (CV, %) and relative error (RE, %) were less than ±15%. Based on this analysis method, the bioequivalence of two cimetidine 400 mg tablet reference (Tagamet® 400mg) and test drug (AK-Cimetidine 400 mg) was evaluated according to the guidelines of Korea Food and Drug Administration (KFDA). Twenty four healthy male volunteers, 66.79±7.98 kg in body weight and 23±2.73 in age, were divided into two groups and a randomized 2×2 cross-over study was employed. After one tablet containing 400 mg of cimetidine was orally administered, blood samples were taken at the predetermined time intervals and the concentrations of cimetidine in plasma were determined by protein precipitation method using HPLC with UV detector. Pharmacokinetic parameters such as AUC and Cmax were calculated and ANOVA was employed for the statistical analysis of parameters. The results were revealed that the differences in AUC and Cmax between two tablets were 1.3 % and 3.4 % respectively. The 90% confidence intervals for these parameters were also within ±20 % (e.g., log 0.9510∼log 1.0259 and log 0.9369∼log 1.1417 for AUC, Cmax, respectively). All of the above mentioned parameters met the criteria of KFDA guidelines for bioequivalence indicating that test drug tablet is bioequivalent to Tagamet® 400 mg tablet.

Physiological and Protein Profiling Response to Drought Stress in KS141, a Korean Maize Inbred Line

김상곤,Hwan Hee Bae,Hwa Jin Jung,Jin-Seok Lee,Jung-Tae Kim,Tae Hoon Go,Beom-Young Son,Seong-Bum Baek,Young-Up Kwon,우미옥,Seonghyu Shin 한국작물학회 2014 Journal of crop science and biotechnology Vol.17 No.4

Understanding the complex response mechanism of a crop to drought is the major step in the developing of tolerant genotypes. Inour study, to investigate physiological traits and proteome dynamics, an inbred maize (Zea mays L.) line (KS141) was subjected to10 days of water-withholding at the V5 or V6 leaf stage. The subsequent analysis of their physiological parameters revealed adecreased relative leaf water content, Fv/Fm, stomatal conductance, net CO2 assimilation rate, leaf transpiration, and water use efficiency,resulting in severe growth retardation of leaf area, stem length and width, aerial part, and root dry matter at 3 and 10 daysafter withholding water. However, aerial part and root dry matter were little changed during drought stress for 3 days. To understandthe proteome dynamics during the 10-day drought stress in maize leaves, comparative proteome analysis was carried out between thewell-watered and drought-treated leaves. Proteins were extracted using phenol extraction method from leaves with/without droughtstress, and then separated by 2-DE. After 2-DE gel analyses, 14 differentially expressed protein spots were identified by MALDITOFmass spectrometry. Out of 14, eleven and three protein spots were found to be up- or down-regulated, respectively. Interestingly, stress-related proteins such as glutathione S-transferase, abscisic stress-ripening proteins, and pathogenesis-related proteinswere increased by drought stress. Our study may provide molecular mechanisms and selective markers for drought tolerantmaize genotypes

An Exploratory, Randomized, Double-Blinded, Placebo-Controlled Study to Evaluate the Efficacy and Safety of Tenofovir with and without Ursodeoxycholic Acid in Patients with Hepatitis B Virus

( Hee Yeon Kim ),( Chang Wook Kim ),( Seung Up Kim ),( Beom Kyung Kim ),( Jung Hyun Kwon ),( Soon Woo Nam ),( Joo Ho Lee ),( Seong Gyu Hwang ),( Hyung Jum Yim ),( Young Kul Jung ),( Jae Youn Cheong ) 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1

Aims: It is suggested that earlier alanine aminotransferase (ALT) normalization during antiviral treatment is associated with a lower risk of hepatic events in patients with chronic hepatitis B (CHB). This study aimed to assess the effect of ursodeoxycholic acid (UDCA) in improving liver inflammation in ALT elevated CHB patients who commence tenofovir therapy. Methods: Eighty-nine tenofovir-naïve patients with CHB were enrolled from 6 centers. Patients were randomly assigned into 3 groups; tenofovir combined with UDCA 1000mg (n=27), UDCA 600mg (n=30), or placebo (n=32), respectively. The primary endpoint is ALT normalization rate at 4 weeks. Results: Out of 89 patients, 10 patients dropped out, 79 patients completed 1-year follow-up. The ALT normalization rates at week 4 by central lab criteria (ALT ≤41 U/L for men, and 33 ≤U/L for women) were 24%, 13.8%, and 23.3% for UDCA 1000mg, UDCA 600 mg, and placebo groups, respectively (P>0.05). ALT normalization rates based on AASLD criteria (ALT ≤30 U/L for men, and 19 ≤U/L for women) at week 4 were not statistically different between 3 groups. ALT normalization rates by central lab criteria between 3 groups did not reach statistical significance at week 12, 24, 36 and 48. However, ALT normalization rates by AASLD criteria was higher in UDCA 1000mg group than UDCA 600mg or placebo groups at week 24, 36 and 48 (P< 0.05). Improvement of liver fibrosis measured by enhanced liver fibrosis score at week 48 was not different between groups (P >0.05). Inhibitory molecules of T cell such as PD-1, CTLA-4 and FoxP3, superoxide dismutase, malondialdehyde and TNF-alpha were checked longitudinally, however, there were no significant differences among these three groups (P>0.05). Conclusions: Combination treatment of UDCA 1000mg with tenofovir can improve ALT normalization rate at 24-48 weeks based on AASLD criteria in ALT elevated CHB patients.

Treatment with Nucleos(t)ide Analogues from HBeAg-Positive Hepatitis Phase is Associated with Lower Risk of Hepatocellular Carcinoma Development than from HBeAg-Negative Hepatitis Phase: A Multicenter Study Involving 10,390 Patients

( Hee Joon Jang ),( Jun Sik Yoon ),( Hwi Young Kim ),( Han Ah Lee ),( Jonggi Choi ),( Seung Up Kim ),( Young-suk Lim ),( Soo Young Park ),( Yeon Seok Seo ),( Na Ryung Choi ),( Minseok Albert Kim ),( H 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1

Aims: Recent studies indicate that the integration of hepatitis B virus (HBV) into host genome, which may directly develop hepatocellular carcinoma (HCC), occurs during HBV e antigen (HBeAg)-positive phase of chronic hepatitis B (CHB). However, it remains still unclear that an early antiviral treatment from HBeAg-positive hepatitis phase can reduce the risk of HCC more profoundly than antivirals from HBeAg-negative hepatitis. This study aimed to investigate the association between HBeAg-positivity and HCC risk in CHB patients who achieved viral suppression using nucleos(t)ide analogues (NAs). Methods: We performed a multicenter study involving 10,390 CHB patients who achieved viral suppression (HBV DNA <2,000 IU/mL) using tenofovir or entecavir from 6 hospitals in Korea from January 2008 to December 2018. The primary endpoint was development of HCC. Death or liver transplantation before HCC development were considered as competing risk events. Index date was set at the first time of viral suppression in each patient. We draw Kaplan-Meier (KM) curves before and after balancing baseline characteristics by inverse probability of treatment weighting (IPTW) according to HBeAg status. Cox regression analyses were performed to find independent predictors of HCC. Analyses were performed in the entire study population, as well as in the two subgroups stratified by the presence of liver cirrhosis (LC). Results: Of the 10,390 patients (median age, 50.9 years; male, 53.7%), 5,094 were HBeAg-positive and 5,296 were HBeAg-negative. During 4.8 years of median follow-up, 1,040 patients (10%) developed HCC: 423 (8.3%) were HBeAg-positive and 617 (11.7%) were HBeAg-negative (modified log-rank test P<0.001; Figure 1A). After adjusting baseline characteristics such as age and presence of LC using IPTW analysis, however, HBeAg status was not associated with HCC risk (modified logrank test P=0.93; Figure 1A). In the non-LC subgroup (n=5,957), HBeAg-positive patients showed a lower rate of HCC development than HBeAg-negative patients before and after IPTW adjustment (both modified log-rank P<0.001; Figure 1B). In the LC subgroup (n=4,433), however, HBeAg-positive patients showed a higher rate of HCC development than HBeAg-negative patients before and after IPTW adjustment (modified logrank P=0.004 for unadjusted KM curves; modified log-rank P=0.009 for adjusted KM curves; Figure 1C). In a multivariable analysis of the entire population, HBeAg-positivity was not an independent predictor of HCC (adjusted hazard ratio [aHR], 0.95; 95% confidence interval [CI], 0.84-1.08; P=0.50) after adjusting variables such as age and presence of LC. In the non- LC subgroup, however, HBeAg-positivity was associated with a significantly lower risk of HCC (aHR, 0.54; 95% CI, 0.42-0.70; P<0.001). In contrast, in the LC subgroup, HBeAg-positivity was associated with a higher risk of HCC (aHR, 1.20; 95% CI, 1.04-1.39; P=0.02). Conclusions: HBeAg status was not an independent predictor of HCC development in the overall CHB patients. In non-cirrhotic CHB patients, however, an earlier treatment with NAs from HBeAg-positive hepatitis phase is associated with a lower risk of HCC occurrence than from HBeAg-negative hepatitis phase. This result supports that an earlier antiviral treatment from HBeAg-positive CHB patients without LC might reduce the risk of HCC more profoundly.

Efficacy and Safety of Tenofovir with and without Ursodeoxycholic Acid in Chronic Hepatitis B Patients with Elevated ALT: Interim Analysis of Phase 2 Randomized, Double-Blinded, Placebo-Controlled Study

( Hee Yeon Kim ),( Chang Wook Kim ),( Ji Young Kim ),( Seung Up Kim ),( Beom Kyung Kim ),( Jung Hyun Kwon ),( Soon Woo Nam ),( Joo Ho Lee ),( Seong Gyu Hwang ),( Hana Park ),( Hyung Jum Yim ),( Young 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: This study aimed to assess the effect of ursodeoxycholic acid (UDCA) in improving liver inflammation in alanine aminotransferase (ALT) elevated chronic hepatitis B (CHB) patients who commence tenofovir therapy. Methods: Eighty-nine tenofovir-naïve patients with CHB were enrolled from 6 centers. Patients were randomly assigned into 3 groups; tenofovir combined with placebo, tenofovir combined with UDCA 600mg, and tenofovir combined with UDCA 1000mg groups. The primary endpoint is ALT normalization rate at 4 weeks. Results: Out of 89 patients, 8 patients dropped out, 58 patients completed 1-year follow-up, and follow-up of remaining patients is ongoing. The ALT normalization rates at week 4 by central lab criteria were 19%, 11%, and 22% for tenofovir monotherapy, UDCA 600mg combination therapy, UDCA 1000mg combination therapy, respectively (P=0.58). ALT normalization rates based on AASLD criteria at week 4 were not statistically different between 3 groups. ALT normalization rates by central lab criteria between 3 groups did not reach statistical significance at week 24 and 48. However, ALT normalization rates by AASLD criteria was higher in UDCA combination group than those in tenofovir monotherapy groups at week 24, 36 and 48 (P< 0.05). Improvement of liver fibrosis measured by enhanced liver fibrosis score at week 48 was not different between groups (P=0.66). Inhibitory molecules of T cell such as PD-1, CTLA-4 and FoxP3, superoxide dismutase, malondialdehyde and TNF-alpha were checked longitudinally, however, there were no significant differences among these three groups (P >0.05). Conclusions: Combination treatment of UDCA with tenofovir can improve ALT normalization rate based on AASLD criteria in ALT elevated CHB patients.

A case of myocardial ischemia due to progression of myocardial bridging

( Kyung Up Kim ),( Jae Kyun Choi ),( Hye Mi Oh ),( Ji Young Woo ),( Hee Su Park ),( Soo Yoon Moon ),( Kyoo Rok Han ) 대한내과학회 2015 대한내과학회 추계학술대회 Vol.2015 No.1

Although myocardial bridging is generally considered benign, it has been associated with myocardial ischemia. A recent analysis of intracoronary ultrasonography showed relaxation of coronary artery is delayed during early diastole in a coronary vessel with myocardial bridging, which can decrease the coronary vasodilator reserve. Furthermore, progression of left ventricular hypertrophy, increased myocardial contractility and tachycardia can make the tunneled segment deeper and systolic compression aggravated. However, there are few reports on the progression of myocardial bridging, which is also related to the myocardial ischemia. We report a case in which progression of myocardial bridging in the mid-left anterior descending artery (LAD) was demonstrated by coronary angiography and caused myocardial ischemia. A 70-year-old man visited the hospital with worsening chestpain. His previous coronary angiography showed myocardial bridging at the mid LAD with 50% luminal narrowing during systole in 2006 (Fig. 1A). His blood pressure was not well controlled because of poor compliance. Follow-up coronary angiography showed myocardial bridging at the mid LAD progressed to severe stenosis (>90% luminal narrowing during systole) and total length of the tunneled artery extended from 22.5 mm to 23.9 mm (Fig. 1C). After blood pressure was controlled, his chest pain was relieved. This is the first report of myocardial ischemia secondary to progression of myocardial bridging caused by sustained hypertension in Korea, which was proved by coronary angiography.

The strategy and potential utilization of temperate germplasm for the improvement of tropical germplasm: genetic diversification program is a GEM (germplasm enhancement of maize) of a resource for(Zea mays L.) growers in USA

Sun-Hee Woo,Tea-Young Oh,Hee-Young Jang,Jung-Hee Ko,Seong-Bum Baek,Young-Up Kwon,Matthew Krakowsky,Major Goodman,Peter J. Balint-Kurti,James B. Holland,Yoon-Sup So 한국육종학회 2013 한국육종학회 심포지엄 Vol.2013 No.07

In U.S.A. maize breeding, exotic germplasm is considered as high-risk and usually introduced by backcrossing specific traits into elite lines. The U.S.A. maize germplasm base is narrow. Only a few open-pollinated varieties are well represented in current programs. Currently, the barrier in using of exotic germplasm in the U.S.A is less formidable than in the 1980s. The major reason is that U.S.A materials are now used in tropical breeding to accelerate earlier maturity and lodging resistance. These exotic materials, developed with U.S.A germplasm, are being introduced back into the U.S.A.Since1994, the ARS-led Germplasm Enhancement of Maize (GEM) project has sought to help broaden the genetic base of America’s corn crop by promising exotic germplasm and crossing it with domestic lines. New hybrids derived from such crosses have provided corn researchers and the producers. These may include improved or alternative native source of resistance to insect pests such as corn rootworms and diseases like northern leaf blight. GEM’s aim is to provide source of useful genetic maize diversity to help the producers to reduce risks from new or evolving insect and disease threats or changes in the environment or respond to new marketing opportunities and demand. During the 2009 growing season, the Ames (Iowa) and Raleigh (North Carolina) locations managed or coordinated evaluations on 17,200 nursery plots as well as 14,000 yield trial plots in Ames and 12,000 in Raleigh. A new “allelicdiversity” study is devoted to exploring and capturing the genetic variation represented by over 300 exotic corn races. Since 2001, GEM has released 221 new corn lines to cooperators for further development into elite commercial new hybrids. GEM has already identified about 50%-tropical, 50%-temperate families tracing primarily to tropical hybrids that are competitive with commercial checks. In North Carolina State University program, they have examined the potential of tropical inbredand hybrids for U.S.A. breeding by crossing temperate-adapted, 100%-tropical lines to U.S.A hybrids. There should be favorably unique alleles or genomic regions in temperate germplasm that can be helpful in tropical maize improvement as well as utilization of tropical lines in temperate areas.

KASL clinical practice guidelines: Management of nonalcoholic fatty liver disease

( Seong Hee Kang ),( Hye Won Lee ),( Jeong-ju Yoo ),( Yuri Cho ),( Seung Up Kim ),( Tae Hee Lee ),( Byoung Kuk Jang ),( Sang Gyune Kim ),( Sang Bong Ahn ),( Haeryoung Kim ),( Dae Won Jun ),( Joon-il C 대한간학회 2021 Clinical and Molecular Hepatology(대한간학회지) Vol.27 No.3