• 한국수정란이식학회지
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• 제27권2호
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• pp.93-100
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• 2012

To investigate the biochemical nature of changes in vaginal physiology during estrus and pregnancy, we examined the cytology and viscosity, and monitored the protein expression profile in vaginal mucus during estrus and pregnancy. The viscosity progressively decreased from estrus to pregnancy. Cell type analysis revealed that white blood cells progressively increased from estrus to pregnancy, while red blood cells progressively decreased during pregnancy. The cornification index (CI) was higher in estrus than in pregnancy. Protein mass spectrumetry identified the presence of ribosome-binding protein 1, GRIP 1 (Glutamate receptor-interacting protein 1)-associated protein 1, DUF729 (Domain of unknown function729) domain-containing protein 1, prolactin precursor, dihydrofolatereductase, and MMP (Matrix metalloprotease)-9 in vaginal mucus. MMP-2 and MMP-9 proteins in the vaginal mucus were active throughout estrus and gestation, as measured by a gelatinase assay, but most abundant in the vaginal mucus on day 0 of estrus. Results from ELISA of MMP-2 and MMP-9 were in accordance with the gelatinase assay. In light of the crucial role of metalloproteinases in extracellular matrix remodeling, the level of MMP-9 in vaginal mucus might be useful as an indicator of estrus and pregnancy to increase the efficiency of reproduction.

• Journal of Ginseng Research
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• 제44권3호
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• pp.496-505
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• 2020

Background: Panax ginseng is a marvelous herbal remedy for all ailments of body. That may be why it is called Panax, which means "cure for all". Melanin is a pigment that gives color to our skin; however, increased melanin production can lead to tumor formation. Human exposure to ultraviolet B radiation has increased extensively owing to the increased sunlight due to global warming. Consequently, a phenomenon called photoaging has been observed for all skin colors and types. As a result of this phenomenon, a set of enzymes called matrix metalloproteinases, which serve as degradation enzymes for extracellular matrix proteins, mainly collagen, is increased, causing depletion of collagen and resulting in early wrinkle formation. Methods: Therefore, in our study, we used the murine melanoma cell line B16/F10 to study the inhibition of melanogenesis by Korean Red Ginseng (KRG) extract in vitro and HRM-2 hairless mice exposed to artificial ultraviolet B to examine the efficacy of KRG in vivo. We prepared a 3% red ginseng extract cream and evaluated its effects on human skin. Results: Our results demonstrated that KRG induced potent suppression of tyrosinase activity and melanin production in B16/F10 cells; moreover, it reduced the transcription and translation of components involved in the melanin production pathway. In the in vivo experiments, KRG potently suppressed the expression of matrix metalloproteinases, reduced wrinkle formation, and inhibited collagen degradation. On human skin, ginseng cream increased skin resilience and skin moisture and enhanced skin tone. Conclusion: Therefore, we conclude that KRG is an excellent skin whitening and antiaging product.

• BMB Reports
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• 제37권3호
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• pp.298-303
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• 2004

In general, a SYPRO Ruby dye is well known as a sensitive fluorescence-based method for detecting proteins by one-or two-dimensional SDS-PAGE (1-DE or 2-DE). Based on the SYPRO Ruby dye system, the combined two-dimensional fibrin zymography (2-D FZ) with SYPRO Ruby staining was newly developed to identify the Bacillus sp. proteases. Namely, complex protein mixtures from Bacillus sp. DJ-4, which were screened from Doen-Jang (Korean traditional fermented food), showed activity on the zymogram gel. The gel spots on the SYPRO Ruby gel, which corresponded to the active spots showing on the 2-D FZ gel, were analyzed by a matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometric analysis. Five intracellular fibrinolytic enzymes of Bacillus sp. DJ-4 were detected through 2-D FZ. The gel spots on the SYPRO Ruby dye stained 2-D gel corresponding to 2-D FZ were then analyzed by MALID TOF MS. Three of the five gel spots proved to be quite similar to the ATP-dependent protease, extracellular neutral metalloprotease, and protease of Bacillus subtilis. Also, the extracellular proteases of Bacillus sp. DJ-4 employing this combined system were identified on three gels (e.g., casein, fibrin, and gelatin) and the proteolytic maps were established. This combined system of 2-D zymography and SYPRO Ruby dye should be useful for searching the specific protease from complex protein mixtures of many other sources (e.g., yeast and cancer cell lines).

• 상하수도학회지
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• 제25권1호
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• pp.85-93
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• 2011

The structural components of microorganism are quite related to the toxin and environmental conditions. The vegetative and dormant cells are quite affected by the physical and chemical environments to survive and they will be dormant when they are in the extreme environment. The mechanism to activate the microorganisms however, is not well defined yet in the area of activation state and sporulation state through the analysis of EPS. Other than that even the main mechanism of prior to acquisition of analysis values is not well understood. Therefore, what kind of specific environment to affect the activation and sporulation will be discussed through the analysis of the extracellular polymeric substances(EPS). EPS are a high molecular weight mixture of polymers presenting both outside of cells and interior of microbial aggregates. They are a major complex materials in microbial aggregation for sustaining them together in a three dimensional matrix. Three commonly used extraction methods were applied to this study their effectiveness and quantification in extracting EPS from several Bacillus microorganisms and activated sludge. Three extraction methods used for this study are regular centrifugation with formaldehyde (RCF), Steaming, and EDTA extraction. The results of EPS contents such as the quantitative proteins, carbohydrates and the ratio of protein versus carbohydrate from the several species with the several specific methods showed in this research. This study aims to get comparable results of the quantitative production of EPS and the effectiveness of sedimentation for Bacillus microorganisms and activated sludge from several wastewater treatment plans. The results revealed that the protein amount extracted was the highest by the Steaming method of three extraction methods before sporulation and the carbohydrate amount extracted was the highest by the RCA method of three extraction methods after sporulation. The higher amount of protein compared with carbohydrate from Bacillus microorganisms affected higher sedimentation efficiency, however it could not be found the relation between the EPS production and sedimentation efficiency for the activated sludge.

• 약학회지
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• 제56권1호
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• pp.9-13
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• 2012

We previously reported that the extract of Taraxacum platycarpum (AF-343) had several biological properties such as skin hydration and anti-inflammatory effects, thereby AF-343 be a promising anti-atopic dermatitis agent. However, few studies have been conducted to evaluate its effect on modulation of extracellular matrix proteins in human skin fibroblasts. The purpose of this study was to investigate the expressions of type I collagen, MMP-1, Smad2/3, and TIMP-1 proteins in AF-343-treated human skin fibroblasts. Human skin fibroblasts were treated by various concentrations of AF-343 (0~2 mg/ml). The expressions of type I collagen, matrix metalloproteinase-1 (MMP-1), Smad2/3, and TIMP-1 proteins were analyzed by Western blot analysis. In addition, level of type I collagen mRNA was analyzed by CAT assay. Expression of type I collagen protein was increased in AF-343-treated human skin fibroblasts by dose and time-dependent manners. Consistent with this result, the expressions of phospho-Smad2/3 in skin fibroblasts were increased and MMP-1 expression was decreased by AF-343 treatment. TIMP-1 expression was not significantly changed in AF-343 treated skin fibroblasts. Extract of Taraxacum platycarpum (AF-343)-induced up-regulation of type I collagen expression was through increased expression of phospho-Smad2/3. These results were occurred combined with down-regulation of MMP-1 in skin fibroblasts. Taken together, this study indicated that AF-343 has property of the modulation of ECM in tissue as well as skin hydration and anti-inflammation.

• Journal of Yeungnam Medical Science
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• 제15권1호
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• pp.55-66
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• 1998

사람과 동일한 증상을 나타내는 동물 모델이 없는 신증후출혈열의 병리 기전을 이해하기 위하여 in vitro culture system을 이용하였다. 신증후출혈열의 주된 병변이 혈관내피세포외에 섬유아세포가 많이 존재하는 신세뇨관 간질 및 폐간질임을 기초로 하여 계태아와 Mongolian gerbil의 섬유아세포에서 fibronectin에 대한 수용체인 ${\alpha}_5{\beta}_1$, integrin에 대한 항체를 처리하여 한탄바이러스 감염에 대한 영향을 관찰하였다. 초대 배양하여 사용한 계태아 섬유아세포 및 MGF 모두 한탄바이러스가 잘 감염되었으나 계태아 섬유아세포에 비하여 MGF에서 바이러스의 증식이 더 우수하였다. 계태아 섬유아세포와 MGF에서 ${\alpha}_5{\beta}_1$, integrin에 대한 항체 처리 5일 후 바이러스의 N 단백의 역가는 항체를 처리하지 않은 대조군에 비하여 각각 32.6%, 72.6%로 virion의 역가는 26.8%, 28.7%로 감소하여 두 세포에서 모두 감염력을 가진 바이러스의 양은 거의 동일하게 감소 되었다. 또한 ${\alpha}_5$ subunit와 ${\beta}_1$, subunit에 대한 항체를 MGF 세포에 처리한 후 바이러스의 N단백의 역가는 65.2%, 59.7%로 ${\alpha}_5{\beta}_1$ integrin에 대한 항체 처리하였을 때(72.6%)와 거의 유사하게 감소하였고 감염력을 가진 바이러스의 양도 26.5%, 29.4%로 감소하여 ${\alpha}_5{\beta}_1$ integrin에 대한 항체 처리시(28.7%)와 거의 유사하게 감소하였다. 이러한 결과는 ${\alpha}_5{\beta}_1$ integrin이 한탄바이러스에 대한 수용체로써 작용할 가능성을 시사하였으며 또다른 기전으로는 ${\alpha}_5{\beta}_1$ integrin의 차단에 의한 이차적인 영향으로 한탄바이러스의 증식에 커다란 영향을 미치는 것을 생각할 수 있었다.

• Journal of Nutrition and Health
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• 제55권2호
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• pp.240-249
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• 2022

본 연구에서는 인간 유방암 세포 라인인 MDA-MB-231 세포에서 GD에 의해 EVs분비 억제에 의한 항암효과를 처음으로 확인하고자 하였다. MDA-MB-231 세포에 GD를 처리하였을 때 농도 의존적으로 세포의 증식률을 억제하는 것을 MTT assay를 통해 확인할 수 있었으며, ROS 염색과 apoptosis marker 단백질인 p-JNK단백질의 증가를 통해 GD에 의한 세포의 증식 억제 효과가 세포사멸에 의한 것임을 유추할 수 있었다. 또한 wound-healing assay, 세포 침윤 및 VEGF 농도를 측정한 결과 GD가 암세포의 이동, 전이 능력을 억제하는 것을 확인하였다. Nanosight를 통해서 MDA-MB-231 세포에서 분비되는 대조군 EVs 및 GD에 의해 변화된 EVs의 사이즈를 확인하였다. 마지막으로 GD를 처리한 MDA-MB-231 세포에서 분비된 EVs보다 GD를 처리하지 않은 대조군에서 분비된 EVs의 단백질 및 particles수가 유의적으로 감소하는 것을 확인을 하였다. 그리고 GD가 MDA-MB-231 세포에서 EVs분비를 감소시키는 것을 대표적인 exosome marker인 TSG101, CD63의 발현 감소로 확인할 수 있었다. 이러한 결과로 인해 GD가 암세포의 EVs 분비를 감소시켜 암세포의 성장 및 전이를 억제하였음을 확인하였다. 본 연구는 GD가 인간 유방암 세포인 MDA-MB-231 세포의 EVs 분비를 억제하는 효과가 있음을 제시하고 있다. 따라서 GD가 유방암의 화학요법 약물로 작용할 수 있음을 시사한다.

• Molecules and Cells
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• 제27권5호
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• pp.503-513
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• 2009

Proteoglycans located in basement membranes, the nanostructures underling epithelial and endothelial layers, are unique in several respects. They are usually large, elongated molecules with a collage of domains that share structural and functional homology with numerous extracellular matrix proteins, growth factors and surface receptors. They mainly carry heparan sulfate side chains and these contribute not only to storing and preserving the biological activity of various heparan sulfate-binding cytokines and growth factors, but also in presenting them in a more "active configuration" to their cognate receptors. Abnormal expression or deregulated function of these proteoglycans affect cancer and angiogenesis, and are critical for the evolution of the tumor microenvironment. This review will focus on the functional roles of the major heparan sulfate proteoglycans from basement membrane zones: perlecan, agrin and collagen XVIII, and on their roles in modulating cancer growth and angiogenesis.

• Korean Journal of Acupuncture
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• 제37권2호
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• pp.88-96
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• 2020

Objectives : The purpose of this study was to investigate the anti-oxidant and anti-aging effect of the seed of Euphorbia lathyris L. extracted by supercritical CO₂. Methods : Human dermal fibroblast cells dosed with the extract from Euphorbia lathyris L. were harvested and the intracellular proteome was analyzed to examine the expression of proteins related collagen synthesis pathway, metalloproteinases (MMPs), extracellular matrix (ECM)-cell interaction, cytokines, and antioxidant enzymes by 2-dimensional gel electrophoresis. Results : Fatty acid analysis of the extract from Euphorbia lathyris L. showed oleic acid was 84% and linoleic acid was 4.1%. Antioxidative effect was about 53% by beta carotene bleaching assay. In 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE) analysis, fifteen protein changes in five mechanisms which were collagen synthesis pathway, MMPs, ECM-cell interaction, cytokines, and antioxidant enzymes were analyzed. Conclusions : This study suggests the supercritical extraction from the seed of Euphorbia lathyris L. could be used as anti-oxidant substances for pharmacopuncture.

• 약학회지
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• 제52권1호
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• pp.48-55
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• 2008

CD29 $({\beta}1-integrins)$ is one of major adhesion molecules involved in regulating cell adhesion, migration and morphological changes. In this study, we investigated the regulatory role of CD29 in monocytic functions using monocytic cell line U937 cells. CD29 was found to be one of highly expressed membrane proteins in U937 cells, according to flow cytometric analysis. The activation of CD29 by agonistic antibody MEM101A and extracellular matrix protein (ECM) fibronectin strongly induced cell-cell and cell-fibronectin adhesions. However, blocking antibodies to CD98 and CD147 showed different inhibitory features in these two adhesion events. Furthermore, U0126, an ERK inhibitor, only blocked cell-cell adhesion but not cell-fibronectin adhesion, indicating that cell-cell or cell-fibronectin adhesion events may be regulated by different molecular mechanisms. Meanwhile, CD29 activation also enhanced ROS generation but not phagocytic ability, and similarly radical scavenger N-acetyl-L-cysteine strongly blocked CD29-mediated cell-cell adhesion, implying that ROS may play a critical role in up-regulating cell-cell adhesion. Therefore, our data suggest that the activation of CD29 may be critically involved in regulating monocytic cell-mediated cell-cell adhesion and ROS generation.