Journal of the korean academy of Pediatric Dentistry
/
v.27
no.2
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pp.309-317
/
2000
Bisphosphonates inhibit bone resorption in vivo and in vitro. Currently proposed mechanism of action of bisphosphonates involves both direct effect on osteoclasts and indirect effect through the mediation of osteoblasts. Recent understanding of molecular mechanism of osteoclastogenesis indicates that osteoclast differentiation is quite tightly regulated by signaling molecules from differentiating osteoblasts. Therefore this investigation was designed to elucidate the effect of bisphosphonate on osteoblast differentation. For this purpose, in vitro effects of etidronate and alendronate on the expression of Cbfa1 a master control gene of osteoblast differentiation, several bone marker genes, and formation of calcified nodules were evaluated. To evaluate the effect of bisphosphonate on calcified nodule formation, osteoblasts isolated from rat calvaria were cultured in a-MEM containing $10^{-4},\;10^{-5},\;10^{-6}M$ of etidronate or $10^{-6},\;10^{-7},\;10^{-8}M$ of alendronate for 15 days, and then stained by alizarin red to determine mineralization. To evaluate the effect of bisphosphonate on osteoblast differentiation, osteoblast cells were cultured in a-MEM containing $10^{-4},\;10^{-5},\;10^{-6}M$ of etidronate or $10^{-6}$ M of alendronate for 8 days. And then total RNA was extracted and northern blot analysis was done to examine the expression of Cbfa1, type I collagen, alkaline phosphatase, osteopontin and osteocalcin. The results were as follows: 1. Etidronate suppressed the calcification of bone nodule in dose dependent manner, while alendronate didn't. 2. The expression of Cbfa1 was decreased dose dependently by etidronate, but increased by alendronate. 3. Etidronate suppressed the expression of type I collagen, osteopontin and osteocalcin in dose dependent manner however alendronate promote the expression of osteoblast marker gene. 4. The expression of alkaline phosphatase was not affected either etidronate nor alendronate. These results suggest that etidronate suppressed the expression of Cbfa1 in dose dependent manner, and consequently the expression of osteoblast marker genes, such as type I collagen, osteopontin and osteocalcin were also suppressed in similar manner. And finally this decreased expression of osteoblastic marker gene prevent calcined bone nodule formation.
Journal of the Korean Society of Food Science and Nutrition
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v.45
no.8
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pp.1090-1098
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2016
The anti-inflammatory effects of ethanol extract from Grateloupia crispata (GCEE) were investigated in lipopolysaccharide (LPS)-stimulated murine macrophages. Anti-inflammatory effects were detected by enzyme-linked immunosorbent assay, Western blotting, and immunohistochemistry. There was no cytotoxic effect on proliferation of macrophages treated with GCEE compared to the control. GCEE significantly inhibited production of pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis $factor-{\alpha}$, and $IL-1{\beta}$] as well as nitric oxide in LPS-stimulated RAW 264.7 cells. In addition, GCEE suppressed expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ in a dose-dependent manner. GCEE significantly reduced activation of mitogen-activated protein kinases. In the in vivo test, evaluation of anti-inflammatory activity of GCEE was performed using croton oil-induced ear edema in ICR mice. Oral administration of 10 mg/kg to 250 mg/kg of GCEE significantly reduced ear edema in a dose-dependent manner compared to croton oil-induced mice. Moreover, GCEE reduced ear thickness and the number of mast cells compared to croton oil-induced mice in the histological analysis. These data suggest that GCEE could be used as a potential source for anti-inflammatory agents.
This research examined the effects of Compositae extract on the inhibition of proliferation and apoptosis in human breast and human gastric cancer cells. Compositae extracts which is used in the experiment are Taraxacum coreanum (TC), Youngia sonchifolia (YS) and Ixeris dentata (ID). The proliferation of SK-BR-3, MDA-MB-231 and AGS cells were investigated by MTT assay. ID and YS extracts inhibited proliferation of SK-BR-3, MDA-MB-231 and AGS cells in a dose-dependent manner, but TC have barely affected. In addition, the most effective extract was ID. To assess the apoptosis of ID extract, the nuclei of human cancer cells were stained with DAPI solution respectively. Chromatin condensation, indicated apoptosis, was increased in a dose-dependent manner. We investigated change of ID extract-induced apoptosis proteins on human cancer cells by western blot analysis. The level of Bcl-2 decreased, whereas the level of Bax, cleaved-PARP increased in dose-dependent manner compared with non-treatment. Also Bax/Bcl-2 ratio, which is used in clinical indicator of apoptosis, was increased at ID extract treatment group compared with non-treatment. Moreover the Bax/Bcl-2 ratio of MDA-MB-231 cell was significantly increased as against SK-BR-3, AGS cells. These results indicated that ID extract have anti-proliferation effect better than YS or TC, and induced apoptosis in human breast cancer MDA-MB-231 cell better than human breast cancer SK-BR-3 cell, human gastric cancer. Even if further research is needed, ID can be developed as a chemopreventive or therapeutic agent of breast cancer.
For veterinary imaging diagnosis, we obtained MR images of the canine brain, spine, kidney and pelvis from 3T MRI system which was equipped with the world first 3T active shield magnet. Spin echo (SE) and fast Spin Echo (FSE) images were obtained from the canine brain, spine, kidney and pelvis of normal and sick dogs using a homemade birdcage and transverse electromagnetic (TEM) resonators operating in quadrature and tuned to 128 MHz. In addition, we employed a homemade saddle shaped RF coil. Typical common acquisition parameters were as follows: matrix=512$\times$512, field of view (FOV)=20cm, slice thickness=3 w, number of excitations (NEX)=1. For T1-weighted MR images, we used TR=500 ms, TE=10 or 17.4 ms. For T2-weighted MR images, we used TR=4000 ms, TE=108 ms. Signal to noise ratio (SNR) of 3T system was measured 2.7 times greater than that of prevalent 1.57 system. The high resolution images acquired in this study represent more than a 4-fold increase in in-plane resolution relative to conventional images obtained with a 20 cm field of view and a 5 mm slice thickness. MR images obtained from 3T system revealed numerous small venous structures throughout the image plane and provided reasonable delineation between gray and white matter The present results demonstrate that the MR images from 3T system could provide better diagnostic quality of resolution and sensitivity than those of 1.5T system. The elevated SNR observed in the 3T high field magnetic resonance imaging can be utilized to acquire images with a level of resolution approaching the microscopic structural level under in vivo conditions. These images represent a significant advance in our ability to examine small anatomical features with noninvasive imaging methods. Moreover, MRI technique could begin to apply for veterinary medicine in Korea.
Journal of the Korean Society of Food Science and Nutrition
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v.44
no.7
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pp.975-982
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2015
Piceatannol (trans-3,4,3',5'-trihydroxystilbene), a natural stilbene, is an analogue of resveratrol. In the present study, possible mechanisms by which piceatannol exerts its pro-apoptotic action in cultured human oral cancer YD-15 cells were investigated. To investigate whether or not piceatannol has effects on cancer cell viability, human oral YD-15 cells were treated with piceatannol (0, 50, and $100{\mu}M$). Piceatannol treatment ($100{\mu}M$) showed the strongest inhibition of cell proliferation and reduced cell viability in a dose-dependent manner. Chromatin condensation detected by DAPI staining significantly increased in a concentration-dependent manner, indicating apoptosis. Piceatannol treatment activated initiator Bax (pro-apoptotic) and cPARP in a concentration-dependent manner. Further, piceatannol induced down-regulation of Bcl-2 (anti-apoptotic). We also evaluated the activity of piceatannol against oral cavity cancer tumors in mice. Piceatannol-treated nude mice bearing YD-15 xenograft tumors exhibited significantly reduced tumor volume and weight due to the potent effect of piceatannol on tumor cell apoptosis, as determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay. Immunohistochemistry staining showed elevated expression of cleaved-caspase-3 as well as reduced expression of Ki-67 in the piceatannol-treated group. Therefore, piceatannol can be developed as a cancer preventive medicine due to its growth inhibitory effects and induction of apoptosis in human oral cancer cells.
Journal of the Korean Society of Food Science and Nutrition
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v.41
no.4
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pp.510-518
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2012
This study examined the biological activity and synergistic effects of an extract of Wa-song ($Orostachys$$japonicus$, OE), a medicinal plant mixture (MPE) and a combination of both at different ratios (1:1, OMPE-1 and 3:1, OMPE-3). Extracts of the medicinal plants mixture were comprised of Baekbokyung, Changchul and Sa-in at the same ratio. The antioxidant activity of the extracts and their complex were tested $in$$vitro$. The $in$$vivo$ antioxidant activity was also analyzed by examining the lipid composition in the liver and kidney of streptozotocin (STZ)-induced diabetes rats. The nitric oxide radical scavenging activity was more than 50% in OMPE-3 at a 1,000 ${\mu}g$/mL concentration. Regarding metal ions, such as $Fe^{2+}$ and $Cu^{2+}$, the antioxidant activity of OMPE-1 and OMPE-3 was higher than that of OE and MPE. OMPE-1 and 3 had higher activity on $Cu^{2+}$ ions than $Fe^{2+}$ ions. The ${\alpha}$-glucosidase inhibition activity of the OE extract was higher than that of MPE and OMPE-1 but the relative activity of OMPE-3 was significantly higher than the others. Freeze-dried MPE, OMPE-1 and OMPE-3 were added to the diet at a level of 1% given to STZ induced diabetes rats for 4 weeks. The OMPE-1 and OMPE-3 administered groups showed significant decreases in the total lipid, total cholesterol and triglyceride levels in the liver and kidney. In these groups, the glycogen accumulation level of the liver was increased significantly. The content of thiobarbituric acid reactive substances (TBARS) in the liver and kidney was decreased but the DPPH radical scavenging activity was increased. These results suggest that Wa-song extract exhibits antioxidant and antidiabetic activity, which are enhanced by a complex with a medical plants extract.
Journal of the Society of Cosmetic Scientists of Korea
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v.30
no.2
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pp.227-233
/
2004
Chronic exposure to solar radiation, particularly ultraviolet (UV) light, causes a variety of adverse reactions on human skin, such as sunburn, photoaging and photocarcinogenesis. Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental facts play critical roles in cellular damage. And, repeated-UV irradiation activated the expression of the matrix metalloproteinase (MMP) and induced skin irritation. Therefore, the development of effective and safe photoprotectants that can reduce and improve the skin damage has been required. The purpose of this study was to investigate the photo-protective effect of several chinese medical plants (Juniperus chinensis) on the UV -induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. UVA induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. Expression of prostaglandin E$_2$ (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay(EIA) using PGE$_2$ monoclonal antibody. In the human skin we tested anti-irritation effect on the SLS-induced damage skin after appling the extract containing emulsion. We found that Juniperus chinensis extract had potent radical scavenging effect by 98% at 100$\mu\textrm{g}$/mL. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25$\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. In the test of proinflammatory cytokines of human keratinocytes Juniperus chinensis extract decreased expression of interleukin 6 about 30%. The amount of PGE$_2$ by HaCaT keratinocytes was significantly increased at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB (p < 0.05). At the concentrations of 3.2-25$\mu\textrm{g}$/mL of this extract, the production of PGE$_2$ by HaCaT keratinocytes (24 h after 10mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p < 0.05). In SLS-induced skin irritation model in vivo, we found to reduce skin erythema and improve barrier recovery after appling Juniperus chinensis extract containing emulsion when compared to irritated non-treated and placebo-treated skin. Our results suggest that Juniperus chinensis extract can be effectively used for the prevention of UV and SLS-induced adverse skin reactions and applied as anti-aging and anti-irritation cosmetics.
Background : Surfactant protein A (SP-A) is important in the regulation of surfactant secretion, synthesis and recycling. SP-A has important roles in regulating surfactant metabolism as well as in determining surfactant's physical properties. Since systemic sepsis is one of the common causes of acute respiratory distress syndrome (ARDS) and abnormalities in surfactant function have been described in ARDS, the authors investigated the effects of endotoxemia on the accumulation of mRNA encoding SP-A and SP-A protein content. Methods : Adult rats were given various doses of intraperitoneal endotoxin from Salmonella enteritidis and sacrificed at different times. SP-A mRNA was measured by filter hybridization method. Lung SP-A protein content was determined by double sandwich ELISA assay using a polyclonal antiserum raised in rabbits against purified rat SP-A. Results : 1) The accumulation of SP-A mRNA in the endotoxin treated group 24 hours after 2mg/kg and 5mg/kg endotoxin treatments was significantly increased 50.9% and 27.3%, respectively, compared to the control group (P<0.001, P<0.025). 2) The accumulation of SP-A mRNA 24 hours in the 5mg/kg endotoxin treated group was significantly increased by 26.5% compared to the control group (P<0.01). 3) Total amount of lung SP-A was not altered at 24 hours by various doses of treatment. Total lung SP-A content 144 hours after endotoxin administration was significantly decreased by 51.4% compared to the control group (P<0.01). Conclusions : The specific regulation of SP-A by various time course in vivo is evident. The late decline in SP-A protein content was unexpected and suggests that SP-A may be differentially regulated during lung inflammation. The functional significance of these alterations remains to be clarified.
This study investigated flavonoid, total phenol, total flavonoid content, antioxidant and antiproliferative activity on human breast cancer cells (MCF-7, MDA-MB-231). Four varieties of Korean green peppers (KP: kkuri pepper, PP: phut pepper, CP: cheongyang pepper, OP: ohi pepper) and one foreign green pepper (JP: jalapeno) were used. The contents of luteolin, quercetin and apigenin, which are abundant flavonoids in green pepper, were the highest in KP. Also, the contents of total phenol, and total flavonoids were the highest in KP, followed by CP, JP, PP, and OP (KP: total phenol $13.29{\pm}0.45$ mg GAE/g D.W., total flavonoid $7.02{\pm}0.13$ mg QE/g D.W. In DPPH ABTS radical-scavenging activity, KP showed the most potent antioxidant activity. In the result of viability in human breast cancer cells, KP had the highest antiproliferative effect. These results suggest that green peppers have significant antioxidant activity and can be a possible candidate for treatment of breast cancer.
Purpose: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models. Methods: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit. Results: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4+ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group. Conclusion: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.
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