• 동의생리병리학회지
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• 제20권2호
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• pp.436-441
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• 2006

Rheumatoid arthritis is a chronic, systemic, and inflammatory autoimmune disorder that affects 1% of the adult population worldwide. Osteoarthritis is a multifactorial disease with high morbidity that is characterized by degradation of the matrix and destruction of articular cartilage. In this study, we examined the inhibition effect of Trachelospermi Caulis on the inflammation($TNF-{\alpha}$, $IL-1{\beta}$, NO), cartilage protection(MMP-13), and cell death in arthritis. RAW 264.7 and SW 1353 cells were cultivated in DMAE(GibcoBRL, USA) with 5% FBS and Fungizone in $37^{\circ}C$, 5% CO2. THP-1 cells were cultivated in RPMI(GibcoBRL, USA) with 5% FBS and Fungizone in $37^{\circ}C$, 5% CO2. Activity of caspase-3, XIAP, Cytochrome C in the cell was examined by using western blot. The results obtained were as Follows; Concentration of nitric oxide in Trachelospermi Caulis treatment group significantly decreased compared with that of non-treatment group (P<0.05). In treated group, Concentration of Trachelospermi Caulis was not significantly associated with cell death. Concentration of $TNF-{\alpha}$ and $IL-1{\beta}$ in Trachelospermi Caulis treatment group decreased significantly compared with that of none treatment group (P<0.05). Relative density of MMP-13 in Trachelospermi Caulis treatment group decreased significantly compared with that of none treatment group and dose-response relationship was observed. After treatment of staurosporin in SW1353 which increases cell death, in Trachelospermi Caulis treated group, the cell death was effectively decreased. In conclusion, these results suggest that Trachelospermi Caulis inhibit inflammation and cell death in arthritis. More researches about effect of Trachelospermi Caulis are considered to need.

• Environmental Analysis Health and Toxicology
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• 제20권3호
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• pp.215-221
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• 2005

Environmental polycyclic aromatic hydrocarbones (PAHs), which are formed during incomplete combustion of fossil fuels, are widely distributed in our environment. Human exposure to PAHs may occur through smoking, polluted air, food consumption and occupational contact. Urinary naphthols, 1-and 2-naphthol, have been suggested as route -specific biomarkers for exposure to airborne PAHs. Cytochrome p450 2E1 (CYP2E1) is known to be a great importance for the metabolism of organic solvents, which is a precacinogens with small molecular weight. This study describes the metabolic differences between PstI and RsaI polymorphisms (c1 allele: PstI-. RsaI+ ; c2 allele: PstI+, RsaI-) of CYP2E1 5-flanking region by genetically modified HepG2 cells, which overexpress the polymorphic regions. The results of CAT assay and western blot in the c2 allele overexpressed cells have higher activities than the cl allele over-expressing cells. However, the metabolism of naphthalene to 2-naphthol has no difference due to the two genotypes. In this study, we established the CYP2E1 polymorphic allele transduced HepG2 cells to screen susceptibility -differences in PAH exposure. In conclusion, the CYP2E1 polymorphism may hardly induce susceptibility differences in PAH exposure monitoring with urinary naphthols.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2011년도 제40회 동계학술대회 초록집
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• pp.265-267
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• 2011

저온 대기압 플라즈마는 21세기에 들어 생의학 분야에 이용될 수 있는 새로운 도구로서 많은 관심을 받고 있다. 대기압 플라즈마는 고가의 진공 장비를 필요로 하지 않고 저전력 구동이 가능하기 때문에 저비용 구동이 가능하고 방전 장치와 전력공급 장치의 소형화에 매우 유리하다. 특히 저온 대기압 플라즈마는 고온의 전자와 저온의 이온 입자가 공존하는 열적 불평형(thermal non-equilibrium) 상태에 있기 때문에 플라즈마의 저온 특성은 유지하면서도 (${\sim}30^{\circ}C$) 물리/화학적 반응성은 매우 높아 그 응용 분야가 매우 넓다. 플라즈마의 다양한 생의학 분야 응용 가운데 세포의 사멸 유발 또는 생장 촉진, 살균/멸균, 지혈, 상처 치유 등에 저온 대기압 플라즈마가 매우 뛰어난 효능을 보인다는 것이 국내외의 다양한 연구를 통해 밝혀지고 있다 [1]. 20 kHz 정현파로 구동되는 플라즈마 장치를 이용한 암 세포 제거 실험에서 플라즈마 처리 효과를 증대시키기 위해 항체-금나노입자 중합체를 암 세포에 주입시켰다 (그림 1(a)). 그 결과 세포의 사멸율은 74%로서 플라즈마 또는 플라즈마-금나노입자만을 처리한 경우에 비하여 사멸율이 매우 높게 나타났다 (그림 1(b)). 이를 통해 암세포 선택성을 가진 항체-금나노입자 중합체와 플라즈마 처리 기술을 융합한 암 세포의 선택적 사멸 유발 기술의 개발 가능성이 열렸다. 또한 플라즈마 처리를 통해 일어나는 세포의 자멸사 기작이 Cytochrome C의 방출 이후 이어지는 Caspase-3의 활성화 경로와 관계가 있음이 밝혀졌다 (그림 1(c)). 치아 미백은 최근 부상하고 있는 저온 대기압 플라즈마의 새로운 응용 분야이다 [5-6]. 대기압에서 동작하는 헬륨 플라즈마 제트를 미백제(과산화수소)와 함께 발치된 치아에 적용하였을 때 (그림 2(a)) 미백제만을 사용하였을 경우에 비해 치아의 색상 변화가 2배 이상 크게 나타나는 것을 확인하였다 (그림 2(b)). 이처럼 최근 그 범위가 크게 넓어지고 있는 저온 대기압 플라즈마의 생의학 응용 기술의 최적화를 위해서는, 다양한 생의학 응용 분야에 따라 요구되는 플라즈마의 특성 및 응용별 기저 기 작에 대한 이해와 연구가 필요하다.

• Molecules and Cells
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• 제39권3호
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• pp.211-216
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• 2016

CYP107W1 from Streptomyces avermitilis is a cytochrome P450 enzyme involved in the biosynthesis of macrolide oligomycin A. A previous study reported that CYP107W1 regioselectively hydroxylated C12 of oligomycin C to produce oligomycin A, and the crystal structure of ligand free CYP107W1 was determined. Here, we analyzed the structural properties of the CYP107W1-oligomycin A complex and characterized the functional role of the Trp178 residue in CYP107W1. The crystal structure of the CYP107W1 complex with oligomycin A was determined at a resolution of $2.6{\AA}$. Oligomycin A is bound in the substrate access channel on the upper side of the prosthetic heme mainly by hydrophobic interactions. In particular, the Trp178 residue in the active site intercalates into the large macrolide ring, thereby guiding the substrate into the correct binding orientation for a productive P450 reaction. A Trp178 to Gly mutation resulted in the distortion of binding titration spectra with oligomycin A, whereas binding spectra with azoles were not affected. The Gly178 mutant's catalytic turnover number for the 12-hydroxylation reaction of oligomycin C was highly reduced. These results indicate that Trp178, located in the open pocket of the active site, may be a critical residue for the productive binding conformation of large macrolide substrates.

• 대한한의학회지
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• 제19권1호
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• pp.251-270
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• 1998

The effects of Gamigunshimtang on the isolated perfused ischemic heart in rats, heart rates, left ventricular pressure, cardiac blood flow and cardiotoxicity were stu.died in H9C2 myoblast cell, myocardial slice culture The results were as follows: 1. The administration of Gamigunshimtang to the rat recovered effectively heart rate, left ventricular pressure and flow rate from the experimental ischemia in perfused rat heart. The release of lactic dehydrogenase after the ischemia also decreased compared to the control group. 2. The administration of Gamigunshimtang to H9C2 myoblast culture enhanced the cell proliferation and protected against doxorubicin and allylamine induced release of the lactic dehydrogenase into the culture medium. It also protected effectively against doxorubicin and allylamine induced decrease of Ca ATPase activity and the increase of NADPH-cytochrome C reductase activity in the microsome. 3. The administration of Gamigunshimtang to the rat myocardial slice culture protected effectively against doxorubicin and allylamine induced decreases of protein synthesis and ATP content, and increases of cvtosolic enzyme, creatin kinase into the medium and lipid peroxidation.

• International Journal of Oral Biology
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• 제45권4호
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• pp.169-178
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• 2020

L-ascorbic acid (L-AA; vitamin C) induces apoptosis in cancer cells. This study aimed to elucidate the molecular mechanisms of L-AA-induced apoptosis in human laryngeal epidermoid carcinoma Hep-2 cells. L-AA suppressed the viability of Hep-2 cells and induced apoptosis, as shown by the cleavage and condensation of nuclear chromatin and increased number of Annexin V-positive cells. L-AA decreased Bcl-2 protein expression but upregulated Bax protein levels. In addition, cytochrome c release from the mitochondria into the cytosol and activation of caspase-9, -8, and -3 were enhanced by L-AA treatment. Furthermore, apoptosis-inducing factor (AIF) and endonuclease G (EndoG) were translocated into the nucleus during apoptosis of L-AA-treated Hep-2 cells. L-AA effectively inhibited the constitutive nuclear factor-κB (NF-κB) activation and attenuated the nuclear expression of the p65 subunit of NF-κB. Interestingly, L-AA treatment of Hep-2 cells markedly activated Akt and mitogen-activated protein kinase (MAPK; extracellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase [JNK]) and and LY294002 (Akt inhibitor), SB203580 (p38 inhibitor) or SP600125 (a JNK inhibitor) decreased the levels of Annexin V-positive cells. These results suggested that L-AA induces the apoptosis of Hep-2 cells via the nuclear translocation of AIF and EndoG by modulating the Bcl-2 family and MAPK/Akt signaling pathways.

• 생명과학회지
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• 제18권11호
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• pp.1499-1506
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• 2008

식용 및 약용으로 이용되는 산초(Zanthoxylum schinifolium)의 줄기로부터 항암활성 성분을 분리하기 위하여, 산초 줄기를 유기용매로 추출하고 각 추출물의 인체 급성백혈병 암세포에 대한 독성 및 세포자살 유도 활성을 조사하였다. Methanol (SS-7), methylene chloride (SS-8), ethyl acetate (SS-9), n-butanol (SS-10)로 추출한 각 시료와 유기용매 추출 후 잔여분획 (SL-14)의 세포 독성을 인체 급성백혈병 Jurkat T 세포주를 대상으로 조사한 결과, 암세포에 대한 세포독성이 주로 methylene chloride 추출분획인 (SS-8)에서 확인되었다. Methylene chloride 추출물 (SS-8)의 Jurkat T 세포주에 대한 세포독성의 기전은 mitochondria로부터cytochrome c 방출, caspase-9 및 caspase-3의 활성화, PARP 분해, internucleosomal DNA fragmentation 등의 일련의 생화학적 반응을 수반하며, 항 세포자살단백질인 Bcl-xL단백질의 과발현에 의해 억제되는 세포자살 기전임을 확인하였다. FADD가 disruption된 Jurkat T cell clone I2.1 ($FADD^{-/-}$) 및 caspase-8가 결핍된 Jurkat T cell clone I9.2 (caspase-$8^{-/-}$)와 함께 the wild-type Jurkat T cell clone A3에 미치는 SS-8의 세포독성작용을 비교 분석한 결과, wild-type Jurkat A3, FADD-deficient Jurkat clone I2.1및 caspase-8-deficient Jurkat clone I9.2 모두는 SS-8의 세포독성에 대해 유사한 정도의 감수성을 나타내었다. 이는 SS-8에 의해 유도되는 apoptosis에 있어서, Fas/FasL system이 관계되지 않음을 시사한다. 한편, SS-8를 GC-MS 분석하여, 9,12-octadecanoic acid (18.62%), 2,4-dihydro-5-methyl-4-(1-methylethylidene)-2-(4-nitrophenyl)- 3H-pyrazol-3-one (14.97%), hexadecanoic acid (14.23%), (z,z)-6,9-pentadecadien-1-ol (13.73%), 5,6-dimethoxy- 2-methyl benzofuran (10.95%), 그리고 4-methoxy-2-methylcinnamic acid (5.38%) 등을 포함한 16가지의 구성 성분과 그 조성비를 확인하였다. 이상의 연구결과는 산초 줄기에 함유된 항암 활성에 대한 규명과 이해를 증진시킨다.

• Molecules and Cells
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• 제28권6호
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• pp.559-563
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• 2009

Glyceraldehyde-3-phosphate is a key intermediate in several central metabolic pathways of all organisms. Aldolase and glyceraldehyde-3-phosphate dehydrogenase are involved in the production or elimination of glyceraldehyde-3-phosphate during glycolysis or gluconeogenesis, and are differentially expressed under various physiological conditions, including cancer, hypoxia, and apoptosis. In this study, we examine the effects of glyceraldehyde-3-phosphate on cell survival and apoptosis. Overexpression of aldolase protected cells against apoptosis, and addition of glyceraldehyde-3-phosphate to cells delayed apoptosis. Additionally, delayed apoptotic phenomena were observed when glyceraldehyde-3-phosphate was added to a cell-free system, in which artificial apoptotic process was induced by adding dATP and cytochrome c. Surprisingly, glyceraldehyde-3-phosphate directly suppressed caspase-3 activity in a reversible noncompetitive mode, preventing caspase-dependent proteolysis. Based on these results, we suggest that glyceraldehyde-3-phosphate, a key molecule in several central metabolic pathways, functions as a molecule switch between cell survival and apoptosis.

• Parasites, Hosts and Diseases
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• 제52권2호
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• pp.197-199
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• 2014

Diphyllobothrium latum and Diphyllobothrium nihonkaiense are the 2 reported main causes of human diphyllobothriasis in the Republic of Korea. However, the differentiation of these 2 species based on morphologic features alone is difficult. The authors used nucleotide sequencing of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene to diagnose Diphyllobothrium spp. Two patients visited the emergency room at Kyungpook National University Hospital on 3 April and 12 April 2013, respectively, with fragments of parasites found while defecating. The parasites were identified as Diphyllobothrium spp. based on morphologic characteristics, and subsequent cox1 gene sequencing showed 99.9% similarity (1,478/1,480 bp) with D. nihonkaiense. Our findings support the hypothesis that D. nihonkaiense is a dominant species in Korea.

• Bulletin of the Korean Chemical Society
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• 제33권10호
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• pp.3233-3240
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• 2012

We demonstrated the combined applications of online protein digestion using trypsin immobilized enzyme reactor (IMER) and dual tandem mass spectrometry with collisionally activated dissociation (CAD) and electron transfer dissociation (ETD) for tryptic peptides eluted through the trypsin-IMER. For the trypsin-IMER, the organic and inorganic hybrid monolithic material was used. By employing the trypsin-IMER, the long digestion time could be saved with little or no sacrifice of the digestion efficiency, which was demonstrated for standard protein samples. For three model proteins (cytochrome c, carbonic anhydrase, and bovine serum albumin), the tryptic peptides digested by the IMER were analyzed using LC-MS/MS with the dual application of CAD and ETD. As previously shown by others, the dual application of CAD and ETD increased the sequence coverage in comparison with CAD application only. In particular, ETD was very useful for the analysis of highly-protontated peptide cations, e.g., ${\geq}3+$. The combination approach provided the advantages of both trypsin-IMER and CAD/ETD dual tandem mass spectrometry applications, which are rapid digestion (i.e., 10 min), good digestion efficiency, online coupling of trypsin-IMER and liquid chromatography, and high sequence coverage.