• 대한수의학회지
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• 제61권3호
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• pp.22.1-22.7
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• 2021

Fenbendazole (FBZ) is a commonly used anthelmintics in veterinary medicine that has recently been found to have anticancer effects in humans. On the other hand, few studies have examined the anti-inflammatory effects of FBZ, and its mechanism is unknown. In this study, mouse bone marrow cells (BMs) were treated with lipopolysaccharide (LPS), a representative inflammation-inducing substance, to generate a situation similar to osteomyelitis in vitro. The effect of FBZ on inflammatory BMs was examined by measuring the metabolic activity, surface marker expression, cell nuclear morphology, and mitochondrial membrane potential (MMP) of BMs. FBZ decreased the metabolic activity and MMP of LPS-treated BMs. Annexin V-fluorescein isothiocyanate/propidium iodide staining and Hoechst 33342 staining showed that FBZ reduced the number of viable cells and induced the cell death of inflammatory BMs. In addition, FBZ reduced the proportion of granulocytes more than B lymphocytes in LPS-treated BMs. Overall, FBZ induces cell death by destabilizing the MMP of LPS-induced inflammatory BMs. In addition to anthelmintic and anticancer agent, FBZ can play a role as an anti-inflammatory agent.

• Bulletin of the Korean Chemical Society
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• 제35권9호
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• pp.2787-2790
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• 2014

Quercetin is a major dietary flavonoid found in onions, apples, tea, and red wine, and potentially has beneficial effects on disease prevention. We carried out this study to investigate the effect of quercetin on UVB-induced matrix metalloproteinase-1 (MMP-1) expression in human keratinocyte HaCaT cells and to further understand the mechanisms of its action. The anti-inflammatory activity of quercetin was investigated and quercetin significantly suppressed the NO production in LPS-stimulated RAW264.7 mouse macrophages. Post treatment of quercetin decreased UV irradiation-induced phosphorylation of JNK, p38 MAPK, and ERK by 91%, 21%, and 17%, respectively. MMP-1 is mainly responsible for the degradation of dermal collagen during the aging process of human skin and quercetin suppressed the UVB-induced MMP-1 by 94%. Binding studies revealed that quercetin binds to p38 with high binding affinity ($1.85{\times}10^6M^{-1}$). The binding model showed that the 4'-hydroxy groups of the B-ring of quercetin participated in hydrogen bonding interactions with the side chains of Lys53, Glu71, and Asp168 and the 5-hydroxy group of the A-ring formed a hydrogen bond with the backbone amide of Met109. The major finding of this study shows that quercetin inhibits phosphorylation of JNK, p38 MAPK, and ERK pathway leading to the prevention of MMP-1 expression in human keratinocyte HaCaT cells. Therefore, our findings suggested the potentials of quercetin as a skin anti-photoaging agent.

• 한국동물생명공학회지
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• 제34권3호
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• pp.247-252
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• 2019

We observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. According to the results from each freezing extender, the sperm membrane integrity (HOST: Hypoosmotic Swelling Test) analysis in TCGGD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Glycerol 3%, Dimethylsulpoxide 3.5 M) is 59.8 ± 0.7, TCGSD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Sucrose 0.1 M, Dimethylsulpoxide 3.5 M) is 59.3 ± 0.5 were significantly higher (p < 0.05) among the experimental groups. And MMPs analysis result, we observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. The expression of active MMP-2 was the highest in sperms frozen in TCGSD and TCGD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Dimethylsulpoxide 3.5 M), Meanwhile, sperms from the TCGGD and TCGED (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Ethylene glycol 3%, Dimethylsulpoxide 3.5 M) group showed lower level of active MMP-2 expression. Together, these results indicate that adding glycerol or sucrose to the sperm freezing buffer would not only suppress MMPs expression but also minimize DNA fragmentation, providing a mean to improve the success rate in the in vitro manipulation of rabbit sperms. Therefore, these results suggest that TCGGD or TCGSD extender method for freezing-thawing of rabbit sperm increased the viability after thawing.

• Toxicological Research
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• 제31권4호
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• pp.363-369
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• 2015

We evaluated the antioxidant activity and anti-wrinkle effects of Aceriphyllum rossii leaf ethanol extract (ARLEE) in vitro using human dermal fibroblasts. The total polyphenol and flavonoid contents of ARLEE were 578.6 and 206.3 mg/g, respectively. At a concentration of $250{\mu}g/mL$, the electron-donating ability of ARLEE was 87.1%. In comparison with the vehicle, ARLEE treatment at $100{\mu}g/mL$ significantly increased type I procollagen synthesis (p < 0.01) by 50.7%. In vitro ARLEE treatment (10 mg/mL) inhibited collagenase and elastase activity by 97.1% and 99.2%, respectively. Compared with the control, ascorbic acid treatment at $100{\mu}g/mL$ significantly decreased matrix metalloproteinase (MMP)-1 protein expression (p < 0.01) by 37.0%. ARLEE treatment at $50{\mu}g/mL$ significantly decreased MMP-1 protein expression (p < 0.01) by 46.1%. Ascorbic acid and ARLEE treatments at $100{\mu}g/mL$ significantly decreased MMP-1 mRNA expression (p < 0.01) by 26.1% and 36.1%, respectively. From these results, we conclude that ARLEE has excellent antioxidant activity and even better anti-wrinkle effects than ascorbic acid in human dermal fibroblasts. These results suggest that ARLEE could be used in functional cosmetics for the prevention or alleviation of skin wrinkles induced by ultraviolet rays.

• 한국축산식품학회지
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• 제35권1호
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• pp.58-70
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• 2015

This study was conducted to examine the effect of egg shell membrane hydrolysates (ESMH) on wrinkle, UV, and moisture protection for cosmetic use. ESMH were fragmented as whole ESMH (before fractioning), Fraction I (> 10 kDa), Fraction II (3-10 kDa), and Fraction III (< 3 kDa). In order to test whether fractionated ESMH can be used for functional cosmetic materials, we examined not only the level of hyaluronic acid and collagen production, but also the MMP-1 activity using a HaCaT and CCD-986Sk cell line. Our study treated each sample of fractionated ESMH with different concentrations (0.01, 0.1, 1 mg/mL). In our in vivo research, we used hairless mice that had been exposed to UV-B to induce wrinkles for 7 wk, then applied Fraction I to the treatment group for 5 wk and then tested skin thickness, minimum erythema dose and moisture content. In addition, Fraction I was high in collagen and HA biosynthesis and it was better than TGF-${\beta}$ in improving of the skin. When TNF-${\alpha}$ caused MMP-1 activity in the CCD-986Sk cells, the whole ESMH and Fraction I proved to be effective in hindering the induction of collagenase depending on the concentration, and also showed outstanding effects in the suppression of skin aging. We found that the treatment group mice's UV-B radiation-induced skin damage was largely mitigated compared to that of the non-treatment group mice. Thus, we have concluded that EMSH helps to mitigate UV-B radiation-induced wrinkles, collagen, HA, MMP-1 activity and can be used for functional cosmetic materials.

• Asian Pacific Journal of Cancer Prevention
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• 제13권5호
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• pp.1793-1797
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• 2012

Objective: To analyze the capacity of neurotrophic artemin to promote the motility and invasiveness of MIA PaCa-2 pancreatic cancer cells. Methods: MIA PaCa-2 was cultured in vitro and studied using transwell chambers for motility and invasiveness on treatment with different concentrations of aArtemin or its receptor $GFR{\alpha}3$ were also determined. Expression of matrix metalloproteinase-2 (MMP-2) and epithelial cadherin (E-cadherin) was quantified using RT-PCR and Western blotting. Results: MIA PaCa-2 pancreatic cancer cell motility and invasiveness was significantly increased with artemin and its receptor $GFR{\alpha}3$ with dose dependence (P<0.01). MMP-2 production was also significantly increased (t = 6.35, t = 7.32), while E-cadherin was significantly lowered (t = 4.27, t = 5.61) (P <0.01). Conclusion: Artemin and its receptor $GFR{\alpha}3$ can promote pancreatic cancer cell motility and invasiveness and contribute to aggressive behavior. The mechanism may be related to increased expression of MMP-2 molecule and down-regulation of E-cadherin expression.

• Journal of Ginseng Research
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• 제43권3호
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• pp.488-495
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• 2019

Background: Timosaponin AIII (TA3) is a steroidal saponin extracted from Anemarrhena asphodeloides. Here, we investigated the anticancer effects of TA3 in MG63 human osteosarcoma cells. TA3 attenuates migration and invasion of MG63 cells via regulations of two matrix metalloproteinases (MMPs), MMP-2 and MMP-9, which are involved with cancer metastasis in various cancer cells. TA3 reduced enzymatic activities and transcriptional expressions of MMP-2 and MMP-9 in MG63 cells. TA3 also inhibited Src, focal adhesion kinase, extracellular signal-regulated kinase (ERK1/2), c-Jun N-terminal kinase (JNK), p38, ${\beta}-catenin$, and cAMP response element binding signaling, which regulate migration and invasion of cells. TA3 induced apoptosis of MG63 cells via regulations of caspase-3, caspase-7, and poly(ADP-ribose) polymerase (PARP). Then, we tested several ginsenosides to be used in combination with TA3 for the synergistic anticancer effects. We found that ginsenosides Rb1 and Rc have synergistic effects on TA3-induced apoptosis in MG63 cells. Methods: We investigated the anticancer effects of TA3 and synergistic effects of various ginseng saponins on TA3-induced apoptosis in MG63 cells. To test antimetastatic effects, we performed wound healing migration assay, Boyden chamber invasion assays, gelatin zymography assay, and Western blot analysis. Annexin V/PI staining apoptosis assay was performed to determine the apoptotic effect of TA3 and ginsenosides. Results: TA3 attenuated migration and invasion of MG63 cells and induced apoptosis of MG63 cells. Ginsenosides Rb1 and Rc showed the synergistic effects on TA3-induced apoptosis in MG63 cells. Conclusions: The results strongly suggest that the combination of TA3 and the two ginsenosides Rb1 and Rc may be a strong candidate for the effective antiosteosarcoma agent.

• 대한본초학회지
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• 제38권4호
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• pp.45-52
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• 2023

Objectives : Fine dust caused by environmental pollution cause oxidative damage and skin aging. In this study, The possibility of using the Codium fragile extract (CFE) as an anti-aging product for skin improvement was evaluated by confirming the protective effect of skin cells from PM10 (particulate matter 10) through inhibition of ROS and MMPs. Methods : In this study, elastase and collagenase inhibitory activities were evaluated. Cell viability was evaluated by treating keratinocytes (HaCaT cell line) with CFE at various concentrations. The cytoprotective effect from PM10 in keratinocyteswas evaluated using the 3-[4,5-dimethylthiazol]-2-yl]-2,5-diphenyl-tetrazoliumbromide (MTT) assay. ROS (reactive oxygen species) was measured in keratinocytes damaged by PM10 using DCF-DA (2′,7′-dichlorofluorescin diacetate) fluorescence staining. As an anti-aging effect of CFE, MMP-1 (matrix metalloproteinase-1) and MMP-1 (matrix metalloproteinase-9) inhibitory activities were evaluated. Results : As a result, CFE decreased the activity of elastase and collagenase. As a result of evaluating the toxicity of CFE, it is non-toxic at a concentration of 10 to 80 ㎍/㎖. Although cell viability of HaCaT cells treated with PM10 decreased, cell viability increased by 38% when treated with CFE 80 ㎍/㎖. Also, ROS decreased by 8.4%, and MMP-1 and MMP-9 decreased at CFE 80 ㎍/㎖. Conclusions : CFE showed excellent cell protection effect, and it is considered that it can be used in anti-aging products for skin improvement by effectively inhibiting ROS and MMPs from keratinocyte damage caused by fine dust.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권5호
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• pp.404-416
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• 2006

Purpose : This study was to clarify the changes in mandibular condyle after unilateral mandibular distraction osteogenesis throughout histological changes and expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2). Materials & Methods : Intraoral distractors were placed via submandibular incision in 8 dogs. Two unoperated animals served as controls. Distraction was performed five days after osteotomy as a rate of 0.5 mm twice per day for 10 days. Two animals were sacrificed on 7, 14, 28, and 56 days after completion of distraction, respectively. Ipsilateral condyles were harvested and processed for histological and immunohistochemical examinations. Results : The condyle cartilage is separated into four layers: fibrous layer, proliferative layer, hypertrophic layer, and calcified layer. At 7 days and 14 days after distraction, the condylar cartilage showed the decreased thickness of the articular cartilage and reduced cellularity. At 28 days after distraction, there was an increase in cellularity of fibrous, proliferative, and hypertrophic layer. However, it demonstrated reduced cellularity compared to the control. At 56 days of after distraction, the articular cartilage was an almost normal histologic structure. Positive Safranin-O staining, indicative of sulfated proteoglycans, was examined in the condylar cartilge of nonloaded control. At 7 days and 14 days after distraction, the sulfated proteoglycans is almost completely depleted from the noncalcified part of the condylar cartilage. At 28 days after distraction, there was an increase in Safranin-O staining intensity. However, the staining intensity of the experimental condyle was weaker than that of the control. At 56 days of after distraction, the condylar cartilage showed almost normal Safranin-O staining pattern. In control condyle, MMP-2 immunostaining was seen in fibrous, proliferative, and hypertrophic layer of condylar cartilage, however, it demonstrated lack of staining in fibrous and proliferative layer. At 7 days and 14 days after distraction, strong MMP-2 immunoreactivity was seen in the fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 28 days after distraction, MMP-2 immunostaining was seen in the fibrous and hypertrophic layer of condylar cartilage, however, their immunoactivity was reduced. At 56 days after distraction, MMP-2 immunoreactivity showed almost normal immunostaining pattern. In control condyle, TIMP-2 immunostaining was primarily seen in fibrous and hypertrophic layer of condylar cartilage, however, it demonstrated lack of staining in proliferative layer. At 7 days after distraction, very weak TIMP-2 immunoreactivity appeared in fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 14 days after distraction, weak TIMP-2 immunoreactivity was seen in the fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 28 days after distraction, TIMP-2 immunoreactivity was increased in the fibrous and hypertrophic layer of condylar cartilage. At 56 days after completion of distraction, TIMP-2 immunoreactivity showed almost normal immunostaining pattern. Conclusions : The results show that short-term outcome of physiologic distraction osteogenesis may lead to degenerative changes in the condylar cartilage. These alterations in the condylar cartilage may be considered as a pressure-related degeneration of the cartilage tissue. However, the long-term results suggest that the condylar cartilage display repair activity after mandibular distraction osteogenesis.

• 한국식품영양과학회지
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• 제42권5호
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• pp.697-704
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• 2013

본 실험에서는 primary culture된 연골세포 in vitro 실험모델과 MIA로 유발한 골관절염 in vivo 실험모델을 이용하여 꾸지뽕나무 잎 추출물의 관절염 예방 효과를 확인하였다. 먼저 MTT 시험법을 통해 세포 사용 적정농도를 $500{\mu}g/mL$ 이하로 결정하여 연골세포사멸 억제를 확인하고, 이를 근간으로 골관절염 동물실험 모델에서 골관절염 예방효과를 확인하였다. $H_2O_2$ 처리에 따른 산화적 독성으로 연골세포 사멸을 유도한 실험에서 꾸지뽕 잎 추출물은 정상세포 수준으로 사멸을 억제하였으며, 이러한 효과는 CTL80의 $200{\mu}g/mL$, CTL10의 $300{\mu}g/mL$ 농도에서 비교적 높게 나타났다. 교원질 합성을 억제하고 분해를 촉진시키는 MMPs(MMP-7, MMP-13)의 발현을 실시간 정량 PCR로 측정하여 발현변화를 살펴보았다. 그 결과 앞선 세포실험 결과와 마찬가지로 CTL80과 CTL10 처리군에서 발현이 유의적으로 낮아졌음을 살펴볼 수 있었다. 특히 CTL80에서 MMP-7과 MMP-13의 발현이 농도 유의적으로 감소하였으며, CTL10의 경우 200, $300{\mu}g/mL$ 농도에서 유의적으로 발현이 감소하는 것을 확인하였다. 세포실험 결과를 바탕으로 동물실험에서의 적정농도를 결정하였으며, 동물독성실험 결과 이상이 없음을 확인하고 실험을 진행하였다. 이때 세포실험결과 선정된 두농도(200, $300{\mu}g/mL$) 간의 차이가 미미하여 동물실험에 적용할 경우 비슷한 실험결과가 나타날 것으로 사료되어 두 실험군 간의 결과를 정확히 구분 짓기 위해 200, $500{\mu}g/mL$ 농도를 선정하여 사용하였다. 골관절염 유발 동물모델을 만들기 위해 SD rat의 관절강에 MIA를 injection 하였으며, 꾸지뽕 잎 에탄올 추출물 투여에 따른 관절염 예방 효과를 관찰하기 위해 관절염 유발 2주일 전부터 1일 1회 경구투여를 실시하고, 유발 후 3주간 지속적으로 투여하고 관찰하였다. 동물 관절의 병리학적 변화를 관찰하기 위하여 Micro-CT 촬영 및 분석을 실시한 결과 Control 군은 골의 강도와 밀도가 감소한 반면, 양성대조군인 MTX 투여군에서 정상군과 비슷한 수준으로 회복된 것을 확인하였고, CTL80-200군과 CTL10-500군에서 Control 군에 비해 유의적으로 수치가 감소하여 골관절염에 따른 손상이 감소한 것을 확인하였다. 동물의 관절조직의 H&E 염색을 통한 조직학적인 변화에서는 골관절염 유발로 연골세포의 손상과 뼈조직의 손상을 관찰하였으며 관절형태를 알아볼 수 없을 정도로 손상된 것을 확인하였다. 반면 CTL80과 CTL10에서는 관절강 세포의 형태가 정상군과 비슷한 둥근모양을 띤 양상을 보였으며 연골조직의 형태가 잘 유지되어 Control 군에 비해 꾸지뽕잎의 투여효과가 나타났음을 관찰하였다. 이상의 결과를 통하여 꾸지뽕 잎 에탄올 추출물은 높은 항관절염 효과가 있을 것으로 사료되며, 항관절염 효능을 지니는 기능성 소재로써 개발 가능성을 확인하였다.