• Ocean and Polar Research
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• 제40권4호
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• pp.249-257
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• 2018

Matrix metalloproteinases (MMPs) are associated with the invasion and metastasis of malignant tumors composed of cancer cells in an increased state of expression. This study evaluates the inhibitory effect of Carex pumila on MMP-2 and MMP-9 activity in phorbol-12-myristate-13-acetate (PMA)-stimulated HT-1080 human fibrosarcoma cells using gelatin zymography, MMPs enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay. C. pumila was extracted twice with dichloromethane ($CH_2Cl_2$) and methanol (MeOH). Treatment with $CH_2Cl_2$ extract and MeOH extract in PMA-stimulated HT-1080 cells effectively reduced the production of MMP-2 and 9. Also, the combined crude extracts ($CH_2Cl_2$ and MeOH) significantly inhibited the enzymatic activities and the expression of MMP-2 and MMP-9 in mRNA and protein levels. The combined crude extracts were partitioned between $CH_2Cl_2$ and water. The organic layer was further fractionated with n-hexane, 85% aqueous methanol (85% aq.MeOH) and the aqueous layer was separated into n-butanol and water, successively. Of the fractions, 85% aq.MeOH fraction showed the highest inhibitory activity of MMP-2 and MMP-9 in gelatin zymography and MMP ELISA kit. Furthermore, 85% aq.MeOH fraction most significantly suppressed cell migration. In RT-PCR and Western blot assay, n-butanol and 85% aq.MeOH fractions exerted the greatest inhibition on mRNA and protein expression of MMP-2 and MMP-9, respectively. As a result, C. pumila can be used as a good anti-invasive agent source.

• Molecules and Cells
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• 제42권2호
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• pp.151-160
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• 2019

Ultraviolet (UV) radiation of the sunlight, especially UVA and UVB, is the primary environmental cause of skin damage, including topical inflammation, premature skin aging, and skin cancer. Previous reports show that activation of nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) in human skin fibroblasts and keratinocytes after UV exposure induces the expression and release of proinflammatory cytokines, such as interleukin-1 (IL-1) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), and subsequently leads to the production of matrix metalloproteases (MMPs) and growth factor basic fibroblast growth factor (bFGF). Here, we demonstrated that TNFR2-SKEE and TNFR2-SKE, oligopeptides from TNF receptor-associated factor 2 (TRAF2)-binding site of TNF receptor 2 (TNFR2), strongly inhibited the interaction of TNFR1 as well as TNFR2 with TRAF2. In particular, TNFR2-SKE suppressed UVB- or $TNF-{\alpha}$-induced nuclear translocalization of activated $NF-{\kappa}B$ in mouse fibroblasts. It decreased the expression of bFGF, MMPs, and COX2, which were upregulated by $TNF-{\alpha}$, and increased procollagen production, which was reduced by $TNF-{\alpha}$. Furthermore, TNFR2-SKE inhibited the UVB-induced proliferation of keratinocytes and melanocytes in the mouse skin and the infiltration of immune cells into inflamed tissues. These results suggest that TNFR2-SKE may possess the clinical potency to alleviate UV-induced photoaging in human skin.

• Journal of Microbiology and Biotechnology
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• 제31권6호
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• pp.794-802
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• 2021

In this study we investigated the role and mechanism of cardamonin on IL-1β induced injury in OA. CHON-001 cells were treated with cardamonin and IL-1β and transfected with silencing nuclear factor erythroid 2-related factor 2 (siNrf2). Cell viability was detected by Cell Counting Kit-8 assay and flow cytometer assay was utilized for cell apoptosis assessment. IL-6, IL-8, TNF-α and Nrf2 mRNA expression was tested by qRT-PCR. Western blot was employed to evaluate MMP-3, MMP-13, Collagen II, Nrf2, NQO-1, NLRP3, Caspase 1 and apoptosis-associated speck-like protein containing a caspase-1 recruitment domain (ASC) protein levels. In CHON-001 cells, IL-1β suppressed cell viability and Collagen II level while promoting cell apoptosis and expression of pro-inflammatory cytokines (IL-6, IL-8, TNF-α), MMPs (MMP-3, MMP-13), NQO-1, and NLRP3 inflammasome (NLRP3, Caspase 1 and ASC), with no significant influence on Nrf2. Cardamonin reversed the effect of IL-1β on cell viability, cell apoptosis, pro-inflammatory cytokines, MMPs, Collagen II, and NLRP3 inflammasome levels. In addition, cardamonin advanced Nrf2 and NQO-1 expression of CHON-001 cells. SiNrf2 reversed the function of cardamonin on IL-1β-induced cell apoptosis and expression of pro-inflammatory cytokines, Nrf2, NQO-1, and NLRP3 inflammasome in chondrocytes. Taken together Cardamonin inhibited IL-1β induced injury by inhibition of NLRP3 inflammasome via activating Nrf2/NQO1 signaling pathway in chondrocyte.

• Nutrition Research and Practice
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• 제15권4호
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• pp.444-455
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• 2021

BACKGROUND/OBJECTIVES: Adipocytes undergo angiogenesis to receive nutrients and oxygen needed for adipocyte' growth and differentiation. No study relating quercetin with angiogenesis in adipocytes exists. Therefore, this study investigated the role of quercetin on adipogenesis in 3T3-L1 cells, acting through matrix metalloproteinases (MMPs). MATERIALS/METHODS: After proliferating preadipocytes into adipocytes, various quercetin concentrations were added to adipocytes, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were performed to evaluate cell proliferation. Glycerol-3-phosphate dehydrogenase (GPDH) activity was investigated as an indicator of fat accumulation. The mRNA expressions of transcription factors related to adipocyte differentiation, CCAAT/enhancer-binding proteins (C/EBPs), peroxisomal proliferatoractivated receptors (PPAR)-γ, and adipocyte protein 2 (aP2), were investigated. The mRNA expressions of proteins related to angiogenesis, vascular endothelial growth factor (VEGF)-α, vascular endothelial growth factor receptor (VEGFR)-2, MMP-2, and MMP-9, were investigated. Enzyme activities and concentrations of MMP-2 and MMP-9 were also measured. RESULTS: Quercetin treatment suppressed fat accumulation and the expressions of adipocyte differentiation-related genes (C/EBPα, C/EBPβ, PPAR-γ, and aP2) in a concentration-dependent manner in 3T3-L1 cells. Quercetin treatments reduced the mRNA expressions of VEGF-α, VEGFR-2, MMP-2, and MMP-9 in 3T3-L1 cells. The activities and concentrations of MMP-2 and MMP-9 were also decreased significantly as the concentration of quercetin increased. CONCLUSIONS: The results confirm that quercetin inhibits adipose tissue differentiation and fat accumulation in 3T3-L1 cells, which could occur through inhibition of the angiogenesis process related to MMPs.

• 한국수산과학회지
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• 제55권6호
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• pp.867-874
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• 2022

Osteoarthritis (OA) is an inflammatory disease due to wear caused by the continuous use of cartilage. Although many drugs for treating OA are being studied, they have side effects, such as digestive disorders and cardiovascular diseases. Glucosamine, a drug derived from natural products, is known to be less effective. Therefore, the marine organism, Sargassum fulvellum, was studied to determine whether it contains substances with a chondroprotective effect on the inflammatory response of chondrocytes induced by interleukin-1β (IL-1β). A 30% ethanol extract of S. fulvellum (SF30%EtOH) has therapeutic and few side effects. We first confirmed the presence of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS), which is expressed during inflammatory reactions. We then examined the expression of collagen type II, which is the main component of the extracellular matrix and cartilage. Finally, the expression of extracellular matrix degrading enzymes, MMPs and ADAMTS-4 and -5, was confirmed. The results showed that SF30%EtOH reduced the expression levels of NO, iNOS, MMPs, and ADAMT-4 and -5, and increased the expression level of collagen type II in chondrocytes induced with IL-1β. Therefore, SF30%EtOH has a chondroprotective effect against inflammation, indicating its potential use for the prevention and treatment of OA.

• International Journal of Industrial Entomology and Biomaterials
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• 제46권2호
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• pp.60-66
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• 2023

Osteoarthritis (OA) is one of the most prevalent degenerative joint diseases and is more common in older and obese individuals. Silkworm male pupae exerts tonic effects by increasing testosterone secretion and the forced swimming time and muscle ratio increased in mice consuming silkworm pupae, which may be beneficial to the older population. Therefore, it will be beneficial to investigate the effects of silkworm pupal extracts (SPE) on OA. To confirm this effect, we prepared SPE in different solvents, and their ability to attenuate matrix metalloproteinases (MMPs) and inflammatory mediators (interleukin-6 [IL-6], interleukin-8 [IL-8] and tumor necrosis factor-α [TNF-α]) were evaluated in an interleukin-1β (IL-1β)-induced SW1353 human chondrosarcoma cell line. 70% ethanolic SPE outperformed the other solvents, reducing MMP-1 and MMP-3 expression by up to 53% and 13%, respectively. Further experiments were performed using 70% ethanolic SPE from three distinct pupation stages in males and females. SPE treatment alleviated MMP-1 expression (43.9-47.4%) regardless of pupation stage and sex. Among the inflammatory mediators, 70% ethanolic SPE alleviated IL-6 and TNF-α levels, and the concentrations thereof were lowest in the early-stage male SPE-treated group (43.15% and 56.74%, respectively). In conclusion, 70% ethanolic SPE may prevent IL-1β-induced osteoarthritis by inhibiting MMPs and inflammatory cytokines. Therefore, SPE is a potential therapeutic agent for the treatment of OA.

• 한국동물생명공학회지
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• 제38권4호
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• pp.254-262
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• 2023

Background: Water deer and sika deer, which breed in the wild environment, are known to have similar reproductive physiology mechanisms. Therefore, this study aimed to analyze the differences in uterine development between water deer and sika deer during estrus. Methods: MMPs and uterine development-related factors were analyzed and morphological differences were compared in the uterus of sika deer captured near Russia near Korea and water deer captured in the wild in Korea. Results: In terms of morphological differences in the uterus, the glands that form villus within the endometrium of the water deer were newly developed, and the formation of small glands was high, but the villus and glands of the sika deer were expanded, and the stroma zone in the myometrium was higher than that of the water deer. Development has increased. Additionally, the expression of PAPP-A and VEGF factors was increased in the endometrium of water deer than in sika deer, but the actions of MMPs were increased in sika deer. Conclusions: As a result of this study, there is a significant difference in the development of glands in the endometrium of water deer and sika deer during estrus, and it is believed that there is a significant difference in the development of the uterus due to the physiological effects of estrus between water deer and sika deer. Additionally, it is believed that there will be differences in the timing at which pregnancy can be decided.

• Nutrition Research and Practice
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• 제18권5호
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• pp.587-601
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• 2024

BACKGROUND/OBJECTIVES: UV radiation is a major factor contributing to DNA damage in skin cells, including stem cells and mesenchymal stem cells, leading to the depletion of these crucial cells. This study examined whether a mixture of Indian gooseberry and barley sprout (IB) could inhibit UVB irradiation and 3-isobutyl-1-methylxanthine (IBMX)-induced photoaging and oxidative stress in the skin using HaCaT, Hs27, and B16F10 cells. MATERIALS/METHODS: The moisturizing-related factors, the collagen synthesis-related c-Jun N-terminal kinase (JNK)/c-Fos/c-Jun/matrix metalloproteinases (MMPs) pathway, and the melanogenesis-related cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/cAMP-responsive binding protein (CREB)/melanocyte inducing transcription factor (MITF)/tyrosinase-related protein (TRP)/tyrosinase activation pathways were analyzed in vitro by an enzyme-linked immunosorbent assay, real-time polymerase chain reaction, and Western blot analysis. RESULTS: The IB complex increased the hyaluronic acid and sphingomyelin levels and the collagenase inhibitory activity, enhanced hydration-related factors, including collagen, hyaluronic acid synthase (HAS), elastin, long chain base subunit 1 (LCB1) (serine palmitoyltransferase; SPT), and delta 4-desaturase sphingolipid 1 (DEGS1), modulated the inflammatory cytokines levels, antioxidant enzyme activities and the NF-κB/MMPs/cyclooxygenase-2 (COX-2) pathway in UVB-irradiated HaCaT cells, and inhibited wrinkle formation by down-regulation of the JNK/c-Fos/c-Jun/MMP pathway and up-regulation of the transforming growth factor-𝛽 receptor I (TGF𝛽R1)/small mothers against decapentaplegic homolog (Smad3)/procollagen type I pathway in UVB-irradiated Hs27 cells. Moreover, the IB complex prevented melanin production by down-regulating the PKA/CREB/MITF/TRP-1/TRP-2 pathway in IBMX-induced B16F10 cells. CONCLUSION: These findings suggest that the IB complex has the potential to serve as a safeguard, shielding the skin from UVB radiation-induced photo-damage.

• Restorative Dentistry and Endodontics
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• 제33권2호
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• pp.148-161
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• 2008

본 연구는 상아질 혼성층의 교원섬유를 가수분해하는 효소인 MMPs (Matrix metalloproteinses)의 억제제로 알려진 chlorhexidine (CHX)을 적용 후 결합강도를 측정하였으며, 이를 각각 열순환 처리 후 결합강도를 측정하였다. 또한 주사전자현미경으로 접착계면에서의 파괴 양상을 비교 분석하였다. 우식이 없는 발거한 32개의 제3대구치의 교합면 상아질을 노출시키고 GI그룹에서는 dentin conditioner를 처리 후 2% chlorhexidine을 적용시키고, 산부식 접착제 그룹에서는 인산 산부식을 시행하고 2% chlorhexidine을 적용 후 3단계 산부식형 상아질 접착제 (Scotchbond Multipurpose, SM), 2단계 산부식형 상아질 접착제 (Single Bond, SB)를 도포하고, 자가부식 접착제 그룹에서는 2% chlorhexidine 적용 후 자가부식 상아질 접착제 (Clearfil Tri-S, TS)를 도포한다. 이후 복합 레진 (Z-250)과 GI (Fuji-II LC)를 충전한 시편을 $1\;mm^2$의 단면을 갖는 beam으로 제작하여 열순환 하지 않거나, 10,000회 열순환 ($5\;{\sim}\;55^{\circ}C$)하였다. Universal testing machine (EZ-test; Shimadzu, Japan)에서 cross head speed 1 mm/min로 인장력을 가하여, 미세인장결합강도를 측정하였다. 실험 결과는 유의수준 0.05 level에서 two-way ANOVA를 이용하여 통계분석하였다. 그 후 파절된 시편의 파괴 양상을 현미경 (SEM)으로 관찰하여 다음과 같은 결론을 얻었다; 1. 2% CHX을 적용한 모든 실험군에서 상아질과의 미세인장결합강도가 증가하였고, 열순환은 상아질과의 미세인장결합강도를 감소시켰다 (P > 0.05). 2. CHX 적용 후 열순환 한 군은 CHX을 적용하지 않고 열순환한 군에 비하여 상아질과의 미세인장결합강도가 높았으며, 특히 GI와 TS군에서 유의한 차이를 나타내었다 (P < 0.05). 3. 파괴 양상 분석 결과, 혼성층에서의 접착성 파괴를 보이며, CHX을 적용하면 혼성층 기저부에서 상부로 파괴 부위가 옮겨가는 양상을 나타내었다. 이상의 연구 결과를 토대로, MMPs 억제제인 2% CHX은 글래스 아이오노머 시멘트와 상아질 접착제의 초기 미세인장결합강도에는 영향을 미치지 않으며, CHX 적용이 접착내구성을 유지하는데 도움이 되었다.

• 한국식품영양과학회지
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• 제42권4호
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• pp.608-614
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• 2013

본 실험에서는 안전하고 우수한 효과를 지닌 천연항산화제의 광노화에 의한 피부주름 형성을 억제하는 천연물질을 탐색하기 위한 연구의 일환으로 CTL80의 식이투여와 로션 도포가 광노화를 유발한 동물모델에서 주름 형성에 미치는 영향을 살펴보았다. 광노화 유발 동물모델을 만들기 위해 hairless mice에 10주 동안 UVB 조사를 주 3회 실시하였다. 형태학적 변화를 관찰한 결과 UVB를 조사한 군은 각질이 증가하고 주름 능선의 두께가 굵고 주름이 깊게 형성한 반면, 양성대조군인 비타민 C와 CTL80 처리한 군에서 주름 형성이 감소하였으며, 특히 2% CTL80 로션 도포군이 가장 주름 형성을 억제하였다. 피부조직의 H&E 염색을 통한 조직학적인 변화에서는 UVB 조사로 두꺼워진 각질층과 표피층을 관찰하였으며 진피층의 교원섬유가 감소되었음이 관찰되었다. CTL80은 각질층과 표피층의 증가를 감소시켰으며 진피층의 교원섬유를 증가시켰음을 관찰하였다. 형태학적 변화와 같이 2% CTL80 로션 도포군의 효과가 가장 양호하게 나타났다. 항산화 상태에 미치는 영향을 살펴보기 위해 동물 혈청의 SOD와 GSH-Px의 활성을 측정한 결과, 비타민C와 CTL80처리 군에서 UVB만 조사한 군에 비하여 활성이 유의적으로 높아졌음을 살펴볼 수 있었다. 항산화 활성 또한 CTL80에 농도 의존적이지는 않았으나 식이 투여군에 비하여 로션 도포군에서 높은 활성을 보였다. 교원질 합성을 억제하고 분해를 촉진시키는 MMPs(MMP-1, MMP-3, MMP-9)의 발현을 실시간 정량 PCR로 측정하여 발현 변화를 살펴보았다. 그 결과 식이 투여군과 로션 도포군 모두 UVB 조사군에서는 발현이 현저히 높아졌으나 비타민 C와 CTL80 처리 군에서 유의적으로 낮아졌음을 살펴볼 수 있었다. 특히 CTL80 로션 도포군에서 MMP-9의 발현이 확연히 낮아졌음을 관찰할 수 있었다. 이러한 결과를 통하여 CTL80은 높은 항산화 활성을 지녔으며 이러한 항산화능을 통하여 UVB로 광노화를 유발시킨 동물모델에서 주름 생성 억제에 긍정적인 영향을 미쳤고 주름 억제 효능을 지니는 기능성 소재로써 개발 가능성을 확인하였다.