• The Korean Journal of Physiology and Pharmacology
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• 제19권3호
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• pp.229-234
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• 2015

Nafamostat mesilate (NM) is a serine protease inhibitor with anticoagulant and anti-inflammatory effects. NM has been used in Asia for anticoagulation during extracorporeal circulation in patients undergoing continuous renal replacement therapy and extra corporeal membrane oxygenation. Oxidative stress is an independent risk factor for atherosclerotic vascular disease and is associated with vascular endothelial function. We investigated whether NM could inhibit endothelial dysfunction induced by tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$ ). Human umbilical vein endothelial cells (HUVECs) were treated with TNF-${\alpha}$ for 24 h. The effects of NM on monocyte adhesion, vascular cell adhesion molecule-1 (VCAM-1) and intracellular adhesion molecule-1 (ICAM-1) protein expression, p38 mitogenactivated protein kinase (MAPK) activation, and intracellular superoxide production were then examined. NM ($0.01{\sim}100{\mu}g/mL$) did not affect HUVEC viability; however, it inhibited the increases in reactive oxygen species (ROS) production and p66shc expression elicited by TNF-${\alpha}$ (3 ng/mL), and it dose dependently prevented the TNF-${\alpha}$ -induced upregulation of endothelial VCAM-1 and ICAM-1. In addition, it mitigated TNF-${\alpha}$ -induced p38 MAPK phosphorylation and the adhesion of U937 monocytes. These data suggest that NM mitigates TNF-${\alpha}$ -induced monocyte adhesion and the expression of endothelial cell adhesion molecules, and that the anti-adhesive effect of NM is mediated through the inhibition of p66shc, ROS production, and p38 MAPK activation.

• 한국임상수의학회지
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• 제25권6호
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• pp.494-500
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• 2008

The aim of this study was to determine the effectiveness of hyaluronic acid-carboxymethylcellulose membrane (GUARDIX-$MB^{(R)}$) barriers on prevention against post-operative peritoneal adhesions. In this study, fourteen mongrel dogs were divided into two experimental groups: 0.1 % hyaluronic acid (0.1HA) group and hyaluronic acidcarboxymethylcellulose membrane (HA-CMC) group. In order to induce adhesions, the anti-mesenteric serosa of the ileum was exteriorized and then abraded in a standard manner by scraping with a scalpel blade to create homogenous petechial hemorrhagic surface over a $1\;{\times}\;1cm$ area. Solution of 0.1HA were simply coated over the abraded tissues, $1.5\;{\times}\;1.5cm$ HA-CMC membrane was placed over the abraded tissues, allowed to spread across the intra-abdominal organs before closure of the abdomen. On day 1 before and day 1, 4, 7, 14, and 21 after operation, venous blood specimens were collected for measurement of fibrinogen and total WBC. The adhesions were blindly assessed 3 weeks later by using a computerized tensiometer. The fibrinogen and total WBC values of two groups showed no statistical significances. The mean tensile strength (gram force, gf) of formed adhesions day 21 after surgery was $88.1\;{\pm}\;55.70gf$ in the 0.1 % HA group and $24.8\;{\pm}\;22.69gf$ in the HA-CMC group. The tensile strength values of adhesion separation HA-CMC membrane group was significantly lower than the 0.1HA group (p<0.05). Therefore, we suggest that HACMC membrane reduce peritoneal adhesions may be applicable to preventing post-operative intraperitoneal adhesions in dogs.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3519-3528
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• 2012

Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1995년도 제3회 추계심포지움
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• pp.107-113
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• 1995

Basement membrane laminin is a multidomain glycoprotein that interacts with itself, heparin and cells. The distal long arm plays major cell and heparin interactive roles. The long arm consists of three subunits (A, B1, B2) joined in a coiled-coil rod attached to a terminal A chain globule (G). The globule is in turn subdivided into five subdomains (Gl-5). In order to analyze the functions of this region, recombinant G domains (rG, rAiG, rG5, rGΔ2980-3028) were expressed in Sf9 insect cells using a baculovirus expression vector. A hybrid molecule (B-rAiG), consisting of recombinant A chain(rAiG) and the authentic B chains (E8-B)was assembled in vitro. The intercalation of rAiG into E8-B chains suppressed a heparin binding activity identified in subdomain Gl-2. By the peptide napping and ligand blotting, the relative affinity of each subeomain to heparin was assigned as Gl> G2= G4> G5> G3, such that G1 bound strongly and G3 not at all. The active heparin binding site of G domain in intact laminin appears to be located in G4 and proximal G5. Cell binding was examined using fibrosarcoma Cells. Cells adhered to E8, B-rAiG, rAiG and rG, did not bind on denatured substrates, poorly bound to the mixture of E8-B and rG. Anti-${\alpha}$6 and anti-${\beta}$1 integrin subunit separately blocked cell adhesion on E8 and B-rAiG, but not on rAiG. Heparin inhibited cell adhesion on rAiG, partially on B-rAiG, and not on E8. In conclusion, 1) There are active and cryptic cell and heparin binding activities in G domain. 2) Triple-helix assembly inactivates cell and heparin binding activities and restores u6131 dependent cell binding activities.

• Journal of Ginseng Research
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• 제43권1호
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• pp.95-104
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• 2019

Background: Oxidized low-density lipoprotein (ox-LDL) causes vascular endothelial cell inflammatory response and apoptosis and plays an important role in the development and progression of atherosclerosis. Ginsenoside compound K (CK), a metabolite produced by the hydrolysis of ginsenoside Rb1, possesses strong anti-inflammatory effects. However, whether or not CK protects ox-LDL-damaged endothelial cells and the potential mechanisms have not been elucidated. Methods: In our study, cell viability was tested using a 3-(4, 5-dimethylthiazol-2yl-)-2,5-diphenyl tetrazolium bromide (MTT) assay. Expression levels of interleukin-6, monocyte chemoattractant protein-1, tumor necrosis factor-${\alpha}$, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1 were determined by enzyme-linked immunosorbent assay and Western blotting. Mitochondrial membrane potential (${\Delta}{\Psi}m$) was detected using JC-1. The cell apoptotic percentage was measured by the Annexin V/ propidium iodide (PI) assay, lactate dehydrogenase, and caspase-3 expression. Apoptosis-related proteins, nuclear factor $(NF)-{\kappa}B$, and mitogen-activated protein kinases (MAPK) signaling pathways protein expression were quantified by Western blotting. Results: Our results demonstrated that CK could ameliorate ox-LDL-induced human umbilical vein endothelial cells (HUVECs) inflammation and apoptosis, $NF-{\kappa}B$ nuclear translocation, and the phosphorylation of p38 and c-Jun N-terminal kinase (JNK). Moreover, anisomycin, an activator of p38 and JNK, significantly abolished the anti-apoptotic effects of CK. Conclusion: These results demonstrate that CK prevents ox-LDL-induced HUVECs inflammation and apoptosis through inhibiting the $NF-{\kappa}B$, p38, and JNK MAPK signaling pathways. Thus, CK is a candidate drug for atherosclerosis treatment.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2008년도 Proceedings of the Convention
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• pp.103-115
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• 2008

Hemodynamic shear stress, the dragging force generated by blood flow, is known as an anti-atherogenic factor. We tested whether lysyl-tRNA synthetase (KRS) will be utilized as an agent controlling shear-sensing systems. KRS was previously known to be secreted as a pro-inflammatory agent. Here we found that KRS inhibited various shear-stimulated signaling pathways. We further found that KRS binds to detergent-resistant membrane (DRM), indicating that KRS binding molecules exist in DRM, specialized regions of the plasma membrane. DRM plays important roles in a variety of cellular processes and consists of gangliosides, signaling molecules and cytoskeletons. We then determined that KRS was colocalized with integrins ${\alpha}4$, ${\alpha}5$ and $av{\beta}3$. In addition, KRS was shown to be associated with sialic acid, existing at the end of gangliosides. Interestingly, the adherent effect of KRS was inhibited by pretreatment with sialic acid. Moreover, treatment of endothelial cells with neuraminidase appeared to inhibit both the KRS adhesion to endothelial cells and shear-stimulated signaling. In conclusion, KRS is likely to be utilized as a vascular regulator.

• Archives of Plastic Surgery
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• 제34권6호
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• pp.765-770
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• 2007

Purpose: Peritendinous adhesion is one of the most notorious complication after the flexor tendon injury. In this study, $Alloderm^{(R)}$(LifeCell Corp., Branchburg, N.J.), which is the decellularized human dermal analogue with its intact native basement membrane components, was used for the prevention of peritendinous adhesions following flexor tendon repair. Methods: Thirty New Zealand white male rabbits were divided equally into 3 groups. In all groups, the flexor digitorum profundus of the third finger of the right back foot was cut totally and repaired by modified Kessler suture technique. Following tendon repair, $Alloderm^{(R)}$ was wrapt around the repaired tendon in the first group and sodium hyaluronate gel was sprayed to the operation field in the second group. In the control group, no external material was applied. The right back foot were immobilized for 6 weeks to optimize the formation of adhesion ingrowth. After death, the third finger that repaired tendons and sheaths was removed en bloc. We checked range of motion. and studied histologically for all groups. Results: The experimental groups had better range of motion than the control group. We checked that the range of motion was 73.5 degrees in $Alloderm^{(R)}$ group, 55.9 degrees in the hyaluronic acid group, and 38.3 degrees in the control group. in the histological study, the experimental group had less adhesions compared with the control group. Conclusion: This study concludes that $Alloderm^{(R)}$ can decrease peritendinous adhesions following flexor tendon repairs in rabbits. We think the method could be used in clinical cases.

• Parasites, Hosts and Diseases
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• 제49권1호
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• pp.79-83
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• 2011

Trichomoniasis is a sexually transmitted disease due to infection with Trichomonas vaginalis, and it can cause serious consequences for women's health. To study the virulence factors of this pathogen, T. vaginalis surface proteins were investigated using polyclonal antibodies specific to the membrane fractions of T. vaginalis. The T. vaginalis expression library was constructed by cloning the cDNA derived from mRNA of T. vaginalis into a phage ${\lambda}$ Uni-ZAP XR vector, and then used for immunoscreening with the anti-membrane proteins of T. vaginalis antibodies. The immunoreactive proteins identified included adhesion protein AP65-1, ${\alpha$-actinin, kinesin-associated protein, teneurin, and 2 independent hypothetical proteins. Immunofluorescence assays showed that AP65-1, one of the identified immunogenic clones, is prevalent in the whole body of T. vaginalis. This study led us to identify T. vaginalis proteins which may stimulate immune responses by human cells.

• Journal of Microbiology and Biotechnology
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• 제26권4호
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• pp.684-692
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• 2016

Acute respiratory virus infectious diseases are a growing health problem, particularly among children and the elderly. Much effort has been made to develop probiotics that prevent influenza virus infections by enhancing innate immunity in the respiratory tract until vaccines are available. Lactobacillus plantarum GB-LP2, isolated from a traditional Korean fermented vegetable, has exhibited preventive effects on influenza virus infection in mice. To identify the molecular basis of this strain, we conducted a whole-genome assembly study. The single circular DNA chromosome of 3,284,304 bp was completely assembled and 3,250 protein-encoding genes were predicted. Evolutionarily accelerated genes related to the phenotypic trait of anti-infective activities for influenza virus were identified. These genes encode three integral membrane proteins, a teichoic acid export ATP-binding protein and a glucosamine - fructose-6-phosphate aminotransferase involved in host innate immunity, the nonspecific DNA-binding protein Dps, which protects bacteria from oxidative damage, and the response regulator of the three-component quorum-sensing regulatory system, which is related to the capacity of adhesion to the surface of the respiratory tract and competition with pathogens. This is the first study to identify the genetic backgrounds of the antiviral activity in L. plantarum strains. These findings provide insight into the anti-infective activities of L. plantarum and the development of preventive probiotics.

• 한국결정성장학회지
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• 제25권5호
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• pp.212-217
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• 2015

본 연구는 해상 양식장 내에 유해 조류로 인해 발생하는 수산 동, 식물의 폐사를 막고, 양식장을 주기적으로 정화하는 폰툰 시스템 내의 멤브레인 층(Membrane Layer)의 생물오손(Biofouling)을 막고자, ML에 적용되는 인공경량골재 및 골재의 구조에 따른 미생물 흡착을 비교하고자 지오폴리머에 은 나노 입자를 코팅하여 생물오손 정도를 확인하고 그 능력을 평가하였다. 이후 코팅에 따른 미생물 기능 억제 능력을 확인하고자 항균 테스트를 실시하였으며, 한국해양과학기술원 남해연구소 앞바다에 설치하여 생물오손 정도와 무게변화율을 관찰하였다. 항균 테스트 결과 은 나노 입자를 코팅한 인공경량골재에서 99.99 %의 세균 감소율을 확인하였으며, 생물오손 정도를 관찰한 결과 2개월부터 은 나노 입자의 효과에 의해 생물오손 정도가 억제되는 것을 확인하였다. 은 나노 입자 코팅에 의한 생물오손 억제기간은 인공경량골재에서 5개월, 지오폴리머에서 3개월 미만으로 나타났으며, 따라서 은 나노 입자가 코팅된 인공경량골재가 효과적으로 생물오손을 감소시킬 수 있는 친환경적이며 효과적인 방법임을 알 수 있었다.