11:10-12:00, Rm 103

OthersU-18-13Sulfuretin inhibits UVB-induced MMP expression by suppressing theNF-κB pathway in human dermal fibroblastsYoung-Rae Lee, Jong-Suk KimDepartment of Biochemistry and Institute for Medical Sciences, Chonbuk National UniversityMedical School, Jeonju, Jeonbuk 560-182, KoreaSulfuretin is one of the main flavonoids produced by Rhus verniciflua. Sulfuretin has beenshown to exhibit many pharmacological activities, including anti-oxidant, anti-obesity, antiinflammatoryand anti-mutagenic activities. However, the anti-skin photoaging effects ofsulfuretin have not yet been reported. In the present study, we investigated the inhibitoryeffects of sulfuretin on MMP-1 and -3 expressions of the human dermal fibroblast cells.Western blot analysis and real-time PCR revealed sulfuretin inhibited UVB-induced MMP-1 and -3 expressions in a dose-dependent manner. UVB strongly activated nuclearfactor-κB (NF-κB) activity, which was determined by IκBαdegradation, nuclearlocalization of p50 and p65 subunit, and NF-κB binding activity. However, UVB-inducedNF-κB activation and MMP expression were completely blocked by sulfuretinpretreatment. In this study, sulfuretin could prevent UVB-induced MMPs expressionsthrough inhibition of NF-κB activation. In conclusion, sulfuretin might be used as apotential agent for the prevention and treatment of skin photoaging.U-18-16Characterization of protocatechuate 3,4-dioxygenase from pseudomonaspseudoalcaligenes kf707Yong-Kweon ChoDepartment of Biochemistry and Health Science, Changwon National University, Changwon,Gyeongnam 641-773, KoreaProtocatechuate 3,4-dioxygenase from pseudomonas pseudoalcaligenes kf707 waspurified by using Q2 IEC and methyl HIC. Optimum temperature and pH were 38℃ and7.0, respectively. Data form pH profiles of the kinetic parameters showed that there aretwo catalytic groups with pKs of 6.2 and 9.4, and two binding groups with pKs of 5.5 and9.0. Dats from chemical modification implied thattyrosine, histidine, cysteine and lysineresidues exist in the active site. Taken together, the residues of cysteine and tyrosine areinvolved in catalysis and that of histidine and lysine in binding.U-18-14Novel pharmacological activities of the roots of Phryma leptostachyavar. asiatica HaraA Rum Park, Hee-Won Choi and Chang-Jin LimDepartment of Biochemistry, Kangwon National University, Chuncheon 200-701, KoreaPhryma leptostachya var. asiatica Hara (Phrymaceae), a perennial, erect, herbal speciesthat usually grows in moist forests, is distributed across Japan, Korea, China, Indochinaand eastern North America. It has been also used as a natural insecticide. Severalinsecticidal lignans have been isolated from the roots of Phryma leptostachya var.asiatica Hara. Until recently, pharmacological activities of Phryma leptostachya var.asiatica Hara have never been clearly assessed. In the present study, the roots ofPhryma leptostachya var. asiatica Hara were extracted with 70% ethanol and dried toproduce the powdered extract, called PLE. PLE exhibited significant anti-angiogenicactivity in the chick chorioallantoic membrane (CAM) assay, one of the representative invivo methods to detect pro-angiogenic and anti-angiogenic activities. It also displayed theinhibitory action on the generation of nitric oxide in the LPS-activated RAW264.7macrophage cells. In the same stimulated macrophage cells, it was also able to decreasethe enhanced reactive oxygen species (ROS) level. It showed antioxidant activity, whichwas detected using the DPPH assay.U-18-17Anti-inflammatory activity of a phospholipid mixture purified fromporcine lung tissues through the suppression of inos and cox-2Hee-Won Choi¹, Jeong-Su Moon², Seong-Hyun Choi²and Chang-Jin Lim¹¹Department of Biochemistry, Kangwon National University, Chuncheon 200-701, and ²BiopidCo. Ltd., Shinbuk, Chuncheon 200-822, KoreaA phospholipid mixture, named KT&G101, was purified from porcine lung tissues and isbeing developed as a remedy for the treatment of atopic diseases. It mainly consists ofphosphatidylcholine (PC) and phosphatidylserine (PS). Its predominant PC component is1,2-dipalmitoylphosphatidylcholine (DPPC). In the previous work, anti-angiogenic, antiinflammatoryand antinociceptive properties of KT&G101 were assessed usingexperimental animal models. In the present work, we tried to further support the antiinflammatoryactivity of KT&G101. KT&G101 decreased the reactive oxygen species(ROS) level elevated by lipopolysaccharide (LPS) in the murine macrophage RAW264.7cells in a concentration-dependent fashion. It was also able to diminish the production ofnitric oxide (NO) in the LPS-stimulated RAW264.7 cells. KT&G101 was identified toexhibit a significant inhibition on the induction of inducible nitric oxide synthase (iNOS)and cyclooxygenase-2 (COX-2) in the LPS-stimulated RAW264.7 cells using westernanalysis. At a concentration of 50 μg/ml, KT&G101 completely suppressed the inductionof iNOS and COX-2 in the stimulated macrophage cells. However, the free radicalscavenging activity of KT&G101 was not assessed in the DPPH assay.U-18-15Monitoring of interferon gamma mRNA expression in whole blood oftuberculosis patients with reverse transcriptase polymerase chainreactionSunghyun Kim¹, Young-Keun Kim², Jang-Eun Cho⁴, Sang-Nae Cho³and HyeyoungLee¹¹Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University,Wonju 220-710, ²Division of Infectious Disease, College of Medicine, Yonsei University WonjuChristian Hospital, Wonju 220-713, ³Department of Microbiology, College of Medicine, YonseiUniversity, Seoul 120-749, and ⁴Department of Biomedical Laboratory Science, Daegu HealthCollege, Daegu 702-722, KoreaTuberculosis (TB) continues to be one of the most critical infectious disease and causes 3million deaths annually. About one-third of world population is latently infected withMycobacterium tuberculosis (Mtb). Recently, new immunodiagnostic tests for TB havebeen developed and called Interferon gamma releasee assay (IGRA). This assay isbased on the host’s immune response to Mtb specific antigens; ESAT-6, CFP-10, TB7.7.Now available commercial IGRA kit such as IFN-γELISA and ELISPOT have showedhigher specificity and sensitivity than conventional tuberculin skin test (TST). However,IGRA test needs higher cost and has some limitations; limited number of samples testedat once and could not completely detect Mtb infection. In this study, therefore, we havedeveloped alternative IGRA using RT-PCR firstly. RT-PCR based IGRA quantify theexpression level of IFN-γmRNA extracted from Mtb specific antigens stimulated wholeblood. Consequently, it is able to reduce turn-around time, cost for test, volume of blood.Furthermore, it could have higher reproducibility and sensitivity than conventional IGRAand TST. For convenience of test procedures, we need to set other quantification methodsuch as quantitative real-time RT-PCR or real-time NASBA.342 Korean Society for Biochemistry and Molecular BiologyU-18-18Piceatannol-3’-O-β-D-glucopyranoside, an arginase inhibitor fromRhubarb, improves vascular function through increase of nitric oxideproductionAinieng Woo and Sungwoo RyooDepartment of Biology College of Natural Sciences, Kangwon National University, Chuncheon200-701, KoreaArginase shares the substrate L-arginine with nitric oxide synthase (NOS), reciprocallyregulates NOS activity by L- arginine bioavailability. Nitric oxide(NOx), which carriesendothelium-derived relaxing factor (EDRF), plays important roles in the maintenance ofvascular homeostasis. Here, we demonstrate that piceatannol-3’-O-β-Dglucopyranoside(PG)has a capacity to improve vascular function via increase of NOproduction. In vascular tension assay, PG treatment to mouse aortic rings increasedacetylcholine-dependent relaxation (vasorelaxant Emax= 104.2 ± 2.685%) compared withuntreated (vasorelaxant Emax = 68.28 ± 6.342%). Furthermore, PG markedly attenuatedthe contractile responses to vasoconstrictor, phenylephrine(PE) and U46619. Themaximal responses (Emax) to PE and U46619 in the presence of PG were 104.6 ±3.303% and 129.2 ± 6.810%, respectively that was comparable to untreated rings (Emaxto PE and U46619, 168 ± 6.02 and 226.6 ± 4.686%). As for EC50, PG appeared lowerEC50 value of 7.314 x 10 -9 M and 3.257 x 10 -8 M, whereas untreated rings was 9.276 x10 -9 M and 2.362 x 10 -8 M against PE and U46619. Hence, PG represents a novelmolecule in developing new strategies for the inhibition and treatment of cardiovasculardiseases derived from endothelial dysfunction.