• 동의생리병리학회지
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• 제21권6호
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• pp.1407-1410
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• 2007

We designed to investigate the relationship the cold-hot theory of herbology and body temperature in experimental model. we prepared four kinds of oriental medicine, which consisted of two cold herbs, Daehwang and Seokgo, and two hot herbs, Buja and Padu. Decrease of body temperature by cold exposure for 12 hour was not inhibited by four herbs oral administration for two weeks. Thermogenesis in mammals is an essential physiological function to maintain the body temperature. Mitochondrial uncoupling proteins(UCPs), which have a potential to generate heat by uncoupling oxidative phosphorylation, apper to play a crucial role in thermogenesis. Therefore UCP is commonly recognized as a key molecule in metabolic thermogenesis and its dysfunction contributes to the development of obesity. In these experiments, Daehwang water extracts inhibited the UCP1 mRNA expression increase by cold exposure in brown adipose tissue. But other herbs did not significantly influence on UCPs mRNA expression in white adipose tissue and seleus muscle tissue. Based on this experiment, we will try to clarify the effects of Daehwang water extracts on UCP1 expression and function.

• IMMUNE NETWORK
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• 제11권6호
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• pp.376-382
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• 2011

Background: Carbon monoxide (CO) is a cytoprotective and homeostatic molecule with important signaling capabilities in physiological and pathophysiological situations. CO protects cells/tissues from damage by free radicals or oxidative stress. NAD(P)H:quinone oxidoreductase (NQO1) is a highly inducible enzyme that is regulated by the Kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway, which is central to efficient detoxification of reactive metabolites and reactive oxygen species (ROS). Methods: We generated NQO1 promoter construct. HepG2 cells were treated with CO Releasing Molecules-2 (CORM-2) or CO gas and the gene expressions were measured by RT-PCR, immunoblot, and luciferase assays. Results: CO induced expression of NQO1 in human hepatocarcinoma cell lines by activation of Nrf2. Exposure of HepG2 cells to CO resulted in significant induction of NQO1 in dose- and time-dependent manners. Analysis of the NQO1 promoter indicated that an antioxidant responsible element (ARE)-containing region was critical for the CO-induced Nrf2-dependent increase of NQO1 gene expression in HepG2 cells. Conclusion: Our results suggest that CO-induced Nrf2 increases the expression of NQO1 which is well known to detoxify reactive metabolites and ROS.

• IMMUNE NETWORK
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• 제9권3호
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• pp.90-97
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• 2009

Background: CD147, as a cellular receptor for cyclophilin A (CypA), is a multifunctional protein involved in tumor invasion, inflammation, tissue remodeling, neural function, and reproduction. Recent observations showing the expression of CD147 in leukocytes indicate that this molecule may have roles in inflammation. Methods: In order to investigate the role of CD147 and its ligand in the pathogenesis of atherosclerosis, human atherosclerotic plaques were analyzed for the expression pattern of CD147 and CypA. The cellular responses and signaling molecules activated by the stimulation of CD147 were then investigated in the human macrophage cell line, THP-1, which expresses high basal level of CD147 on the cell surface. Results: Staining of both CD147 and CypA was detected in endothelial cell layers facing the lumen and macrophage-rich areas. Stimulation of CD147 with its specific monoclonal antibody induced the expression of matrix metalloproteinase (MMP)-9 in THP-1 cells and it was suppressed by inhibitors of both ERK and NF-${\kappa}B$. Accordingly, the stimulation of CD147 was observed to induce phosphorylation of ERK, phosphorylation-associated degradation of $I{\kappa}B$, and nuclear translocation of NF-${\kappa}B$ p65 and p50 subunits. Conclusion: These results suggest that CD147 mediates the inflammatory activation of macrophages that leads to the induction of MMP-9 expression, which could play a role in the pathogenesis of inflammatory diseases such as atherosclerosis.

• Journal of Microbiology and Biotechnology
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• 제25권8호
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• pp.1281-1290
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• 2015

Thermolysin and its homologs are a group of metalloproteases that have been widely used in both therapeutic and biotechnological applications. We here report the identification and characterization of a novel thermolysin-like protease, BtsTLP1, from insect pathogen Bacillus thuringiensis serovar Sichuansis strain MC28. BtsTLP1 is extracellularly produced in Bacillus subtilis, and the active protein was purified via successive chromatographic steps. The mature form of BtsTLP1 has a molecule mass of 35.6 kDa as determined by mass spectrometry analyses. The biochemical characterization indicates that BtsTLP1 has an apparent K_m value of 1.57 mg/ml for azocasein and is active between 20℃ and 80℃. Unlike other reported neutral gram-positive thermolysin homologs with optimal pH around 7, BtsTLP1 exhibits an alkaline pH optimum around 10. The activity of BtsTLP1 is strongly inhibited by EDTA and a group of specific divalent ions, with Zn²⁺ and Cu²⁺ showing particular effects in promoting the enzyme autolysis. Furthermore, our data also indicate that BtsTLP1 has potential in cleaning applications.

• 한국정보전자통신기술학회논문지
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• 제13권5호
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• pp.397-402
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• 2020

타입 1 콜라겐 단백질은 인체 내에서 가장 많이 존재하는 단백질이다. 이 단백질은 점탄성 거동을 보이며 이는 힘줄에서도 찾아볼 수 있다. 분자동역학 시뮬레이션 방법에는 rescaling 방법과 reassignment 방법으로 온도를 조절할 수 있다. rescaling 방법은 온도를 주어진 온도로 책정하는 방법이고, reassignment 방법은 원하는 온도로 맥스웰 분포를 이용하여서 온도를 책정하는 방법이다. 우리는 reassignment 방법에서 콜라겐 단백질의 거동이 시간에 따라서 변화하는 현상을 찾아내었다. 반면에 rescaling 방법에서는 시간에 무관하게 거동하였다. 콜라겐에 다른 속도로 인장을 가하였을 경우, 예를 들어 0.5, 1, 2, 5 Å/ps의 속도로 40 Å까지 힘을 가했을 경우, rescaling 방법에서는 속도에 따른 변화가 거의 없었던 반면, reassignment 방법의 경우 대략 80nm, 100nm, 130nm, 180nm까지 인장이 되었음을 보여준다. 이 현상에 대한 물리학적 의미를 명확하게 규명하지는 못하였지만, 단백질에 관한 시뮬레이션을 실행하는데 있어서 주의를 기울여 수행하여야 한다는 점에서 이 논문의 가치가 있다고 생각한다.

• The Plant Pathology Journal
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• 제18권6호
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• pp.313-316
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• 2002

Systemic virus symptoms caused by a Potexvirus were observed on leaves of infected hosta (Hasta spp.) plants cultivated in Seoul, Korea. Symptoms on diseased hosta plants include mosaic, mottle, irregular blotchy patches, and chlorotic spots on or distortion of the leaves. No other viruses, such as Cucumber mosaic virus, Lily symptomless virus, or Potyvirus, were detected from the same plants by electron microscopy and by Western blot and RT-PCR analyses, indicating that they were singly infected by the potexvirus. The symptoms differed among cultivars and species of hosta, and affected the quality of plants for commercialization, as well as, plant growth and flowering of susceptible cultivars. Most of the cultivars and species investigated were susceptible to the virus, while some were not infected by the virus at all. Purified virus particles were of filamentous type with unaggregated forms 540 nm in length, which is a typical potexviral morphology. The virus consisted of a single-stranded RNA molecule of 6 kb long for genome and single component of coat protein (CP) about 27 kDa. The CP strongly reacted with the antiserum against Hosta vims X (HVX), suggesting that the virus is an isolate of HVX. This is the first report of the occurrence and identification of HVX from hosta plants in Korea.

• 동의생리병리학회지
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• 제34권3호
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• pp.126-135
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• 2020

The purpose of this study was to investigate the antioxidant, anti-inflammatory and anti-cellular adhesion molecules effects of Allii Macrostemonis Bulbus, Artemisiae Capillaris Herba, Curcumae Radix, Crataegi Fructus, Salviae Militiorrhizae Radix complex extract(AMCP) on the inhibition of atherosclerosis in HUVEC. We measured DPPH radical scavenging activity and ABTS radical scavenging activity of AMCP to evaluate its antioxidant effect. And we also measured the expression level of NF-κB, IκBα, ERK, JNK, p38 proteins to evaluate its anti-inflammatory effect. Lastly, we measured the expression level of MCP-1, ICAM-1, VCAM-1 mRNA and their level to evaluate its anti-celluar adhesion molecules. AMCP did not show any cytotoxicity in HUVEC within the concentraion tested except for a concentration of 400 ㎍/㎖. AMCP increased the DPPH radical scavenging activitiy and ABTS radical scavenging activity in HUVEC as the concentration of AMCP rises. AMCP significantly reduced NF-κB, IκBα, JNK, ERK and p38 protein expression in HUVEC compared to control group. AMCP significantly reduced MCP-1, ICAM-1, VCAM-1 gene expresion in HUVEC compared to control group. AMCP significantly decreased the levels of MCP-1, ICAM-1, VCAM-1 in HUVEC compared to control group. These results suggest that AMCP has effects on antioxidation, anti-inflammation and anti-cellular adhesion molecule, which helps the treatment and prevention of dyslipidemia and atherosclerosis.

• Toxicological Research
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• 제30권4호
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• pp.261-266
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• 2014

Environmental pollutants such as polycyclic aromatic hydrocarbons (PAHs) have been implicated in cancer development and progression. However, the effects of PAHs on carcinogenesis are still poorly understood. Here, we characterized a mouse cancer cell line BNL 1ME A. 7R.1 (1MEA) derived by transformation of non-tumorigenic liver cell line BNL CL.2 (BNL) using 3-methylcholanthrene (3MC), a carcinogenic PAH. RT-PCR and immunoblot analysis were used to determine the expression level of mRNA and proteins, respectively. To determine functionality, cell motility was assessed in vitro using a transwell migration assay. Both mRNA and protein levels of E-cadherin were significantly decreased in 1MEA cells in comparison with BNL cells. While the expression levels of mesenchymal markers and related transcription factors were enhanced in 1MEA cells, which could lead to increase in cell motility. Indeed, we found that 7-day exposure of BNL cells to 3-MC reduced the level of the adhesion molecule and epithelial marker E-cadherin and increased reciprocally the level of the mesenchymal marker vimentin in a dose-dependent manner. Taken together, these results indicate that the process of epithelial-mesenchymal transition (EMT) may be activated during premalignant transformation induced by 3-MC. A mechanism study to elucidate the relation between 3-MC exposure and EMT is underway in our laboratory.

• Biomolecules & Therapeutics
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• 제28권5호
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• pp.482-489
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• 2020

G protein-coupled receptor kinase 5 (GRK5) has been considered as a potential target for the treatment of heart failure as it has been reported to be an important regulator of pathological cardiac hypertrophy. To discover novel scaffolds that selectively inhibit GRK5, we have identified a novel small molecule inhibitor of GRK5, KR-39038 [7-((3-((4-((3-aminopropyl)amino)butyl)amino)propyl)amino)-2-(2-chlorophenyl)-6-fluoroquinazolin-4(3H)-one]. KR-39038 exhibited potent inhibitory activity (IC₅₀ value=0.02 µM) against GRK5 and significantly inhibited angiotensin II-induced cellular hypertrophy and HDAC5 phosphorylation in neonatal cardiomyocytes. In the pressure overload-induced cardiac hypertrophy mouse model, the daily oral administration of KR-39038 (30 mg/kg) for 14 days showed a 43% reduction in the left ventricular weight. Besides, KR-39038 treatment (10 and 30 mg/kg/day, p.o.) showed significant preservation of cardiac function and attenuation of myocardial remodeling in a rat model of chronic heart failure following coronary artery ligation. These results suggest that potent GRK5 inhibitor could effectively attenuate both cardiac hypertrophy and dysfunction in experimental heart failure, and KR-39038 may be useful as an effective GRK5 inhibitor for pharmaceutical applications.

• Korean Journal of Acupuncture
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• 제27권1호
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• pp.31-47
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• 2010

Objectives: To investigate the anti-inflammatory effects of cultivated wild ginseng pharmacopuncture in lipopolysaccharide (LPS)-induced inflammatory rat model. Methods: Sprague-Dawley rats were divided into 4 groups; LPS control (n=6), LPS+cultivated wild ginseng pharmacopuncture at CV4 (n=6), LPS+cultivated wild ginseng pharmacopuncture at CV17 (n=6), and LPS+cultivated wild ginseng pharmacopuncture at Ex-HN1 (n=6). Pharmacopuncture (0.1 ml) was given every two days for 4 weeks followed by inflammation induction by peritoneal LPS injection (5 mg/kg). Blood, liver tissue, and peritoneal lavage fluid were taken and proinflammatory cytokines and other related factors were analysed. Results: Compared with the control group, CV4 and Ex-HN1 pharmacopuncture groups significantly attenuated plasma IL-$1{\beta}$, IL-6, and TNF-$\alpha$ increase at 2h and 5h after LPS injection (P<0.05). A significant difference from control group emerged at 5 h for plasma IL10 (P<0.05). For liver cytokines analyzed at 5 h after LPS injection, only CV4 pharmacopuncture group showed significant difference in TNF-$\alpha$ and IL-10 (P<0.05). Blood CD4/CD8 ratio and the phagocytic activities of polymorphonuclear neutrophils were not different from those of control group in all pharmacopuncture groups (P>0.05). CV4 pharmacopuncture significantly attenuated increase of plasma ${NO_3}^-/{NO_2}^-$, Intracellular adhesion molecule-1 (ICAM-1), cytokine-induced neutrophil chemoattractant-1 (CINC-1), and prostaglandin $E_2$ ($PGE_2$) compared with the control group (P<0.05). Monocyte chemoattractant protein-1, $PGE_2$, and CINC-1 level of CV4 pharmacopuncture group was significantly different from those from the control group (P<0.05). Conclusions: These results indicate that cultivated wild ginseng pharmacopuncture at CV4 may have a potent anti-inflammatory effect in an LPS-induced inflammatory rat model.