• Journal of Ginseng Research
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• 제41권3호
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• pp.257-267
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• 2017

Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.

• Journal of Periodontal and Implant Science
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• 제27권4호
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• pp.883-908
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• 1997

Bone remodeling is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. The process is tightly regualted at the local level by an incompletely known netwotk of peptide and non-peptide fators. Nitric oxide(NO), synthesized by nitric oxide synthetase(NOS) from L-arginine, is becoming recognized as an important bioregualtory molecule in a variety of tissue, but little is known about its possible role in periodontal tissue. The purpose of this study is to investigate the expression of nitric oxide synthetase(NOS) in inflamed gingiva and the effects of cytokine on the expression of NOS protein. The expression of NOS in gingival tissue was evaluated by immunohistochemical staining for $NOS_1$, $NOS_2$, $NOS_3$. The effect of cytokine on the expression of NOS in human periodontal ligament cells and osteoblast-like HOS cells by western blot analysis. Further, we studied that NO functions in periodontal ligament cells as a regulatory molecule. PDL cells incubated with NOS inhibitor and donor. The protein expression, type I collagen & non-collagenous protein, nitrate production and cell proliferation were evaluated The results were as follows. 1. $NOS_1$, $NOS_2$, $NOS_3$ was rarely distributed in healthy gingiva, but stronger stained in gingival epithelium, endothelial cells, and mononuclear cells of inflammed gingiva. 2. The cytokine stimulated $NOS_1$, and $NOS_3$ protein were not inducing or inhibitory effect to compared with control in PDL and HOS cells. 3.Incubation of cells with combination of $TNF-{\alpha}$, $IFN-{\gamma}$, LPS result in a time dependant increase in $NOS_2$ expression, reaching a maximal level after 24 hours of stimulation. 4. The osteonectin protein inhibitory effect of NMA, inhibitor of NOS, was reversed by Larginine in dose dependant manner. 5. NMA decreased cell poliferation and nitrate production, but the inhibitory efffect of NMA was also prevented by the NO donor, sodium nitropruiside. These results suggest that exogenously synthesized NO was playing a stimulating effect on cell proliferation or on non-collagenous protein expression. Therefore NO have an important role in mediation of localized bone destruction associated inflammatory bone disease such as periodontitis.

• 대한물리치료과학회지
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• 제6권3호
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• pp.63-82
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• 1999

Recent studies have revealed that Nitric oxide(NO) was one of the demonstration for the physiological regulator, endothelial derived relaxing factor(EDRF) and that NO was produced by ultraviolet irradiation in human. Thus, the present author have carried out a experimental study on the change of hematological, histological value of ultraviolet irradiation in sprague-dawley rats. The subjects were divided into four groups of ten rats each selected at random. There were 4 groups: 1. no irradiation control; 2. ultraviolet $75mJ/cm^2$ irradiation group; 3. ultraviolet $150mJ/cm^2$ irradiation group; 4. ultraviolet $225mJ/cm^2$ group. After a irradiation, hematological and histological tests were performed to observe erythrocyte, hemoglobin, hematocrit, MCH, MCHC, MCV, $O_2$ saturation, pH, $PO_2,\;PCO_2$ value and to observe histological changes. In hematological tests, erythrocyte, hemoglobin, hematocrit significantly increased in $75mJ/cm^2$ than control group and more $150mJ/cm^2$ ultraviolet irradiation group respectively. Also In blood gas tests, $PO_2$ significantly increased in $75mJ/cm^2$ and more $150mJ/cm^2$ group than control group. Whereas $PCO_2$ significantly decreased in $75mJ/cm^2$ and more $150mJ/cm^2$ group than control group (Duncan-Tukey test, P<0.05). In histological tests, control and $75mJ/cm^2$ group unchanged, but more $150mJ/cm^2$ group changed that it was cytolysis, cytotoxic effect, acanthosis, proliferation of keratinocyte, appearance of amorphous cell and pyknotic nucleus, production of sunburn cell. In conclusion, the present author results support the importance of the relation between NO effect and hematological, histological value by ultraviolet B irradiation.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2503-2509
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• 2012

Considerable research has been conducted concerning galectin-9 and carcinomas, but little information is available about any relation with the hepatocellular carcinoma. In this study, we employed a small interfering RNA (siRNA) targeting galectin-9 to down-regulate the expression in HepG2 cells. As a result, after galectin-9 expression was reduced, cell aggregation was suppressed, while other behaviour such as the proliferation, adhesion and invasion to ECM, cell-endothelial adhesion and transendothelial invasion of the cells were markedly enhanced. When tumors of 200 patients with hepatocellular carcinoma were tested for galectin-9 expression by immunohistochemistry, binding levels demonstrated intimate correlations with the histopathologic grade, lymph node metastasis, vascular invasion and intrahepatic metastasis (P<0.05). Moreover, survival analysis indicated that patients with galectin-9 expression had much longer survival time than those with negative lesions, and the Log-rank test indicated that this difference was statistical significant (P<0.0001). The Cox proportional hazards model suggested that negative galectin-9 expression in hepatocellular carcinoma represented a significant risk factor for patient survival. We propose that galectin-9 might be a new prognostic factor with antimetastatic potential in patients with hepatocellular carcinoma.

• Asian Pacific Journal of Cancer Prevention
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• 제15권1호
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• pp.167-174
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• 2014

Tumor angiogenesis, growth and metastasis are three closely related processes. We therefore investigated the effects of barbigerone on all three in the B16F10 tumor model established in both zebrafish and mouse models, and explored underlying molecular mechanisms. In vitro, barbigerone inhibited B16F10 cell proliferation, survival, migration and invasion and suppressed human umbilical vascular endothelial cell migration, invasion and tube formation in concentration-dependent manners. In the transgenic zebrafish model, treatment with $10{\mu}M$ barbigerone remarkably inhibited angiogenesis and tumor-associated angiogenesis by reducing blood vessel development more than 90%. In vivo, barbigerone significantly suppressed angiogenesis as measured by H and E staining of matrigel plugs and CD31 staining of B16F10 melanoma tumors in C57BL/6 mice. Furthermore, it exhibited highly potent activity at inhibiting tumor growth and metastasis to the lung of B16F10 melanoma cells injected into C57BL/6 mice. Western blotting revealed that barbigerone inhibited phosphorylation of AKT, FAK and MAPK family members, including ERK, JNK, and p38 MAPKs, in B16F10 cells mainly through the MEK3/6/p38 MAPK signaling pathway. These findings suggested for the first time that barbigerone could inhibit tumor-angiogenesis, tumor growth and lung metastasis via downregulation of the MEK3/6/p38 MAPK signaling pathway. The findings support further investigation of barbigerone as a potential anti-cancer drug.

• 대한의생명과학회지
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• 제25권1호
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• pp.107-112
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• 2019

The corpus luteum (CL) is composed to various cells, such as luteal steroidogenic cells (LSCs), luteal thecal steroidogenic cells (LTCs), luteal endothelial cells (LECs), fibroblast, immune cells and blood cells. The life span of CL is controlled by proliferation and apoptosis of luteal cells. Therefore, this study investigated apoptotic factors in luteal cells derived from bovine CL. The CL tissues were collected from bovine ovaries and luteal cells were isolated from middle phase CL. Then, LTCs and LECs were separated according to cellular morphology from LSCs. The expression of Bax, Bcl-2, Fas and tumor necrosis factor 1 receptor (TNF1R) mRNA and protein were analyzed using quantitative RT-PCR and western blot. Results show that, Bax and TNFR1 mRNA expression were significantly increased at late group than early and middle groups, otherwise Bcl-2 were significantly decreased at late group than early group (P<0.05). Fas mRNA expression were significantly decreased in middle group compared to early and late groups (P<0.05). In addition, Bax and Bcl-2 mRNA in LTCs was lower than LSCs, Fas mRNA was higher than LSCs. The Bcl-2 protein expression was lower at LTCs than LSCs, especially Fas protein in LTCs was significantly lower than LSCs and LECs (P<0.05). Otherwise, TNFR1 protein of LTCs were similar with LSCs but higher compared with LECs. In conclusion, we suggest that the results may help understanding of apoptosis ability in luteal cells according to cell type during CL regression of estrous cycle.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9319-9325
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• 2014

Alkaloids are the most extensively featured compounds of natural anti-tumor herbs, which have attracted much attention in pharmaceutical research. In our previous studies, a mixture of major three alkaloid components (5, 6-dihydrobicolorine, 7-deoxy-trans-dihydronarciclasine, littoraline) from Hymenocallis littoralis were extracted, analyzed and designated as AHL. In this paper, AHL extracts were added to human liver hepatocellular cells HepG-2, human gastric cancer cell SGC-7901, human breast adenocarcinoma cell MCF-7 and human umbilical vein endothelial cell EVC-304, to screen one or more AHL-sensitive tumor cell. Among these cells, HepG-2 was the most sensitive to AHL treatment, a very low dose ($0.8{\mu}g/ml$) significantly inhibiting proliferation. The non-tumor cell EVC-304, however, was not apparently affected. Effect of AHL on HepG-2 cells was then explored. We found that the AHL could cause HepG-2 cycle arrest at G2/M checkpoint, induce apoptosis, and interrupt polymerization of microtubules. In addition, expression of two cell cycle-regulated proteins, CyclinB1 and CDK1, was up-regulated upon AHL treatment. Up-regulation of the Fas, Fas ligand, Caspase-8 and Caspase-3 was observed as well, which might imply roles for the Fas/FsaL signaling pathway in the AHL-induced apoptosis of HepG-2 cells.

• Tuberculosis and Respiratory Diseases
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• 제62권1호
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• pp.43-50
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• 2007

배경 및 목적: 인체암에서 C-erbB-2의 과발현에 대한 광범위한 연구가 시행되고 있지만, 임상적 의의에 대해서는 논란의 대상이 되고 있다. 악성종양에서 신생혈관형성은 암의 진행에 필수적인 요소이다. 본 연구에서는 비소세포 폐암에서 C-erbB-2의 과발현의 의미, 암조직의 신생혈관형성에 중요한 역할을 하는 VEGF의 발현 및 두 표지자의 연관성을 연구하였다. 방 법: 파라핀 포매된 95예의 비소세포 폐암조직을 이용하여 avidin-biotin complex 법에 의한 면역조직화학적 염색을 시행하였으며, 연구재료는 원발성 편평세포암종 60예 그리고 원발성 선암종 35예를 대상으로 하였다. 결 과: 비소세포 폐암에서 선암종의 C-erbB-2의 과발현은 편평세포암종보다 의미 있게 높게 나타났다. 임상병기에 따른 C-erbB-2와 VEGF의 과발현은 진행암에서 증가하는 경향을 나타내었으나, 초기암과의 유의한 차이는 없었다. 선암종에서 C-erbB-2와 VEGF의 발현은 유의하게 상관성을 나타내었다. 결 론: 비소세포 폐암에서 C-erbB-2의 과발현은 암의 유형에 따라 다르다는 것을 알 수 있었으며, C-erbB-2의 과발현이 높게 나타나는 선암종에서는 이 성적과 암세포의 VEGF의 발현이 상관성을 나타낸다는 것을 알 수 있었다.

• Macromolecular Research
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• 제9권2호
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• pp.107-115
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• 2001

Poly(L-lactide-co-glycolide)(PLGA) was blended with poly[$\omega$-methacryloyloxyethyl phospho-rylcholine-co-ethylhexylmethacrylate (PMEH)] (PLGA/PMEH) to endow with new functionality i.e., to improve the cell-, tissue- and blood-compatibility. The characteristics of surface properties were investigated by measurement of contact angle goniometer, Fourier-transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR) and electron spectroscopy for chemical analysis (ESCA). NIH/3T3 fibroblast and bovine aortic endothelial cell were cultured on control and PLGA/PMEH surfaces for the evaluation of ceil attachment and proliferation in terms of surface functionality such as the concentration of phosphoryl-choline. Also, the behavior of platelet adhesion on PLGA/PMEH was observed in terms of the surface functionality. The contact angles on control and PLGA/PMEH surfaces decreased with increasing PMEH content from 75$^{\circ}$ to about 43$^{\circ}$. It was observed from the FTIR-ATR spectra that phosphorylcholine groups are gradually increased with increasing blended amount of MPC. The experimental P percent values from ESCA analysis were more 3.28∼7.4 times than that of the theoretical P percent for each blend films. These results clearly indicated that the MPC units were concentrated on the surface of PLGA/PMEH blend. The control and PLGA/PMEH films with 0.5 to 10.0 wt% concentration of PMEH were used to evaluate cell adhesion and growth in terms of phosphorylcholine functionality and wettability. Cell adhesion and growth on PLGA/PMEH surfaces were less active than those of control and both cell number decreased with increasing PMEH contents without the effect of surface wettability. It can be explained that the fibronectin adsorption decreased with an increase in the surface density of phosphorylcholine functional group. One can conclude the amount of the protein adsorption and the adhesion number of cells can be controlled and nonspecifically reduced by the introduction with phosphorylcholine group. Morphology of the adhered platelets on the PLGA/PMEH surface showed lower activating than control and the number of adhered platelets on the PLGA/PMEH sample decreased with increasing the phosphorylcholine contents. The amount of fibrinogen adsorbed on the PLGA/PMEH surface demonstrated that the phospholipid polar group played an important role in reducing protein adsorption on the surface. In conclusion, this surface modification technique might be effectively used PLGA film and scaffolds for controlling the adhesion and growth of cell and tissue, furthermore, blood compatibility of the PLGA was improved by blending of the MPC polymer for the application of tissue engineering fields.

• 한국식품영양과학회지
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• 제46권6호
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• pp.671-680
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• 2017

본 연구 결과 검은콩 물 에탄올 추출물과 Lactobacillus rhamnosus GG(LGG)와 Bifidobacterium animals subsp. lactis BB-12(BB-12)를 이용하여 발효시킨 발효검은콩 물 에탄올 추출물의 발효 전과 후의 성분변화를 분석하고, 검은콩과 검은콩 발효 추출물이 모유두 세포 성장에 미치는 효과를 알아보기 위해 primary 인간 모유두 세포(HFDPC)에 검은콩 추출물(BWE, BEE)과 발효검은콩 추출물(BWE-F, BEE-F)을 다양한 농도로 처리하여 세포독성을 확인하였으며, 이를 바탕으로 적정 처치 농도를 결정하여 모발 성장촉진(VEGF와 KGF/FGF7)과 억제($TGF-{\beta}1$과 AR)에 관여하는 유전자 발현을 mRNA 수준에서 확인하였다. 나아가 검은콩 추출물이 HFDPC의 생존 촉진에 미치는 영향을 Akt와 Erk의 인산화 활성을 통해 비교 분석하였다. LGG와 BB-12를 이용하여 발효시킨 검은콩은 발효가 진행됨에 따라 pH, 총폴리페놀, 총당 환원당의 함량이 감소하였으며, 검은콩 4종 추출물(BWE, BEE, BWE-F, BEE-F) 중 BWE, BEE, BWE-F가 VEGF의 mRNA 발현을 증가시키고, 모든 처리군에서 KGF/FGF7의 mRNA 발현을 유의적으로 증가시킴을 확인할 수 있었다. 나아가 BWE, BEE, BWE-F가 Erk의 활성을 증가시켰음을 확인할 수 있었다. 따라서 검은콩 물 추출물과 검은콩 에탄올 추출물, 그리고 발효검은콩 물 추출물이 인간 모유두 세포의 세포성장에 모발 성장 촉진인자의 활성과 Erk의 활성화 등을 통한 기전으로 긍정적인 영향을 미침으로써 모발 성장 및 모발 건강을 위한 기능성원료로서의 가능성을 확인할 수 있었으며, 유산균 2종(LGG, BB-12)을 이용한 발효검은콩이 검은콩과 비교하여 모발 성장 촉진 관련 단백질 발현에서는 유의적인 우월성을 가지지는 않음을 확인하였다. 추후 다른 유산균 균 총을 이용한 발효검은콩의 연구와 더불어 보다 정밀한 발효를 통한 검은콩 추출물의 성분과 조성의 변화를 바탕으로 한 모발 성장 및 모주기 관련 연구가 이루어져야 할 것이라 생각된다.