• Journal of Applied Biological Chemistry
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• 제63권1호
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• pp.75-82
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• 2020

본 연구에서는 한국에서 육종한 신육성품종 그린볼 사과 껍질 추출물(GBE)이 피부 광노화 인자조절에 대한 억제효과를 확인하였다. 피부에서 광노화 인자 조절에 대한 억제 효과를 확인하기 위해 CCD986sk fibrobalst cell에 UVB로 광노화를 유도시킨 후 세포에 GBE를 처리하였다. 광노화 인자 조절 효과를 측정한 결과 GBE가 COL1A2, MMP-1, MMP-9, TIMP-1 단백질 발현량에서 UVB에 의해 자극된 MMP-1, MMP-9 단백질 합성을 억제하였으며, COL1A2, TIMP-1 단백질의 경우 발현량이 유의적으로 증가하는 것을 확인하였다. Photoaging-related factors인 COL1A2, MMP-1, MMP-9, HAS2, TGF-β, TIMP-1의 mRNA 발현량을 측정한 결과 GBE에 의해 MMP-1, MMP-9 단백질 발현을 효과적으로 억제하였으며, MMPs와 type procollagen 발현에 관여하는 조절인자인 TIMP-1과 TGF-β의 발현량이 증가함에 따라 COL1A2의 생성량이 증가하는 것을 확인할 수 있었다. 또한 피부를 구성하는 구조 단백질 중 하나인 hyaluronic acid 생성에 관여하는 HAS2의 발현량 증가도 확인하였다. 따라서 GBE는 광노화의 인자 억제 조절에 대한 우수한 효능을 가졌으며, 피부 노화를 예방하는 기능성 소재로서 활용이 가능할 것으로 판단되었다.

• 분석과학
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• 제24권2호
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• pp.135-141
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• 2011

세포외 기질의 주요 단백질 구성요소들을 가수분해시켜 재편성(turnover)과 재형성(remodeling)에 핵심 역할을 하는 효소가 matrix metalloproteinases (MMPs)이다. MMPs 중에서 MMP-2와 MMP-9는 catalytic domain이 fibronectin-like domain에 의해 hemopexin-like domain 부위와 떨어져 있는 점이 다른 MMP들과 다르다. MMP-9 억제제의 개발로 간암전이를 막을 수 있다고 보고되고 있다. Hovenia dulcis Thunberg에서 MMP-9의 활성을 저해하는 물질을 분리 정제하였고, 분리된 물질에 대한 MMP-9의 활성억제 여부를 확인하였다. Ethyl acetate (EA)에 의해 용출 분리된 두개의 화합물(화합물 A, 화합물 B)과 MeOH로 용출 분리된 한 개의 화합물(화합물 C)에서 MMP-9의 활성에 저해를 보였고, $^1H$와 $^{13}C$ NMR, GC-MS 그리고 IR로 이들의 구조를 분석하였다. 화합물 A는 hydroxyl기와 methoxyl기로 치환된 벤젠고리를 함유하는 화합물로 catechine 계열로 추정되었으며, 화합물 B와 C는 hydroxyl기와 methoxyl기로 치환된 벤젠 고리와 분자내 carbonyl기를 갖고 있는 nobiletin 계열의 화합물로 추정되었다. 그리고 화합물 A는 MMP-9 활성을 1.0%농도에서 76% 억제하였으며, 화합물 B는 동일한 농도에서 66% 억제하는 것으로 나타났다. 그리고 화합물 C는 1.0%농도에서 71% 억제하는 것으로 나타나 화합물 A가 MMP-9의 활성 저해능이 가장 좋은 것으로 나타났다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권5호
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• pp.402-407
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• 2006

Vascular endothelial cells release proteinases that degrade the extracellular matrix (ECM), thus enabling cell migration during angiogenesis and vasculogenesis. Sildenafil citrate stimulates the nitric oxide-cyclic guanosine monophosphate pathway through inhibition of phosphodiesterase type V (PDE5). In this report, we examined the mechanisms underlying sildenafil citrate-induced cell migration using cultured mouse aortic endothelial cells (MAECs). Sildenafil citrate induced migration and proteinase secretion by murine endothelial cells. Sildenafil citrate induced the secretion of matrix metalloproteinase-2 (MMP-2) and MMP-9, which is inhibited by $NF-{\kappa}B$ inhibitors. Sildenafil citrate also induced the secretion of plasmin, which is inhibited by PI 3'-kinase inhibitors. It is suggested that sildenafil citrate-induced migrating activity in endothelial cells may be accomplished by increased secretion of proteinases.

• 한국해양바이오학회지
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• 제10권2호
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• pp.83-90
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• 2018

Ultraviolet (UV) radiation causes inflammation and matrix metalloproteinase (MMP) over-expression and extra cellular matrix depletion, leading to skin photo-aging such as wrinkle formation, dryness, and sagging. In this study, we demonstrated that pretreatment with the hexane extract of microalgae protects UVB mediated cell damages. The results of clinical study showed that Microalgal Oil treated group reduced wrinkle and improve elasticity. All these results suggest Microalgal Oil may be useful as new photo-aging cosmetics for protection against UV induced activity.

• Asian Pacific Journal of Cancer Prevention
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• 제14권10호
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• pp.6025-6030
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• 2013

Background: Matrix metalloproteinase-2 (MMP-2) is an enzyme with proteolytic activity against matrix proteins, particularly basement membrane constituents. A single nucleotide polymorphism (SNP) at -1306, which disrupts a Sp1-type promoter site (CCACC box), results in strikingly lower promoter activity with the T allele. In the present study, we investigated whether this MMP-2 genetic polymorphism might be associated with susceptibility to colorectal cancer (CRC) in the Saudi population. We also analyzed MMP-2 gene expression level sin CRC patients and 4 different cancer cell lines. Materials and Methods: TaqMan allele discrimination assays and DNA sequencing techniques were used to investigate the $C^{-1306}T$ SNP in the MMP-2 gene of Saudi colorectal cancer patients and controls. The MMP-2 gene expression level was also determined in 12 colon cancer tissue samples collected from unrelated patients and histologically normal tissues distant from tumor margins. Results and Conclusions: The MMP-2 $C^{-1306}T$ SNP in the promoter region was associated with CRC in our Saudi population and the MMP-2 gene expression level was found to be 10 times higher in CRC patients. The MMP-2 $C^{-1306}T$ SNP is significantly associated with CRC in the Saudi population and this finding suggested that MMP-2 variants might help predict CRC progression risk among Saudis. We propose that analysis of this gene polymorphism could assist in identification of patient subgroups at risk of a poor disease outcome.

• 대한치과교정학회지
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• 제28권6호
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• pp.915-926
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• 1998

Cytoskeleton은 세포핵과 세포외 기질을 연결하고 있어서 기질에 가해지는 물리적 힘에 의해 cytoskeletal change가 유도되고 이에 의해 세포의 개조활성이 영향을 받는다고 생각되어 왔다. 본 연구는 골모세포 활성에 대한cytoskeletal change의 역할을 규명하기 위한 것으로서, 신생 백서로부터 조골세포양 세포를 분리, 배양하고 네가지 농도의 cytochalasin B(CB) 또는 colchicine(COL)을 3시간 처리하였다. 다시 배양액을 교환하고 24시간 동안 배양하여 prostaglandin $E_2\;(PGE_2)$, interleukin-6(IL-6), tumor necrosis factor-$\alpha$(TNF-$\alpha$) 및 matrix metalloproteinase-1 (MMP-1) 생산을 측정하고 통계적으로 비교하였으며 cytoskeletal protein actin 변화를 관찰하기위하여 면역형광염색하고 형광현미경으로 관찰하여 다음과 같은 결과를 얻었다: 1. CB 처리군에서 $PGE_2$ 생산이 증가되는 경향을 보였고 COL 처리군에서는 약물농도에 비례하여 증가하였다. 2. IL-6 생산은 CB농도 1.0 ${\mu}g/ml$일때를 제외하고 증가되었다. 3. TNF-$\alpha$도 CB 농도가 1.0 ${\mu}g/ml$ 일때를 제외하고 증가하였다. 4. MMP-1 생산은 CB 처리군에서 감소하는 경향을 보이고 COL 처리군에서는 변화되지 않았다. 5. CB처리군에서는 cytoskeletal actin stress fibers가 사라지고 세포모양이 둥글어지는 경향을 보였다. 이상의 결과로 미루어 보아 cytoskeletal rearrangement는 골모세포유사세포의 활성, 특히 $PGE_2$, IL-6, 및 TNF-$\alpha$같은 paracrine/autocrine factor의 생산과 관련있는 것으로 보인다.

• Restorative Dentistry and Endodontics
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• 제30권2호
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• pp.138-144
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• 2005

Enterococcus faecalis의 추출물을 성인의 말초혈액으로부터 얻은 인간의 백혈구를 이용해 Matrix Metalloproteinase-8 (MMP-8)의 생산에 미치는 영향에 대해서 알아보았다. Enterococcus faecalis를 배양한 뒤 초음파 분쇄를 하여 추출물을 얻어냈다. 이중 일부는 적정한 농도의 Ca(OH)_2$를 처리하였다. 이후 E. faecalis 추출물군, E. faecalis 추출물에 $Ca(OH)_2$ 를 처리한 군으로 분리하여 다형핵백혈구를 자극하여 MMP-8의 생산량을 측정하였다. MMP-8는 처리하지 않은 PMN group에서도 나타났고, PMN을 E. coli LPS로 자극시킨 경우 증가하였다 (p < 0.05) SEF group 들에서는 MMP-8의 생성량이 SEF 의 농도가 증가함에 따라 감소하였다 (p < 0.05). 따라서 Enterococcus faecalis는 PMN의 MMP-8 생산 억제 효과를 가진다고 볼 수 있다. $Ca(OH)_2$로 처리한 SEF 군의 경우 각기 다른 SEF 농도에서도 모두 MMP-8 생성량이 아무 처리도 하지 않은 PMN 군보다 많았다 (p < 0.05) . 하지만 각 군들 사이에 통계적 차이는 없었다 (p > 0.05).

• The Korean Journal of Physiology and Pharmacology
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• 제20권2호
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• pp.221-228
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• 2016

We investigated whether prunetin affects the proteolytic activity, secretion, and gene expression of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as in vivo production of MMP-3 in the rat knee joint to evaluate the potential chondroprotective effect of prunetin. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcriptionpolymerase chain reaction (RT-PCR) was used to measure interleukin-$1{\beta}$ (IL-$1{\beta}$)-induced expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), and ADAMTS-5. In rabbit articular chondrocytes, the effects of prunetin on IL-$1{\beta}$-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of prunetin on MMP-3 protein production was also examined in vivo. The results were as follows: (1) prunetin inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5; (2) prunetin inhibited the secretion and proteolytic activity of MMP-3; (3) prunetin suppressed the production of MMP-3 protein in vivo. These results suggest that prunetin can regulate the gene expression, secretion, and proteolytic activity of MMP-3, by directly acting on articular chondrocytes.

• Clinical and Experimental Pediatrics
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• 제56권3호
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• pp.116-124
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• 2013

Purpose: Tumor necrosis factor (TNF)-${\alpha}$ is thought to contribute to pulmonary hypertension. We aimed to investigate the effect of infliximab (TNF-${\alpha}$ antagonist) treatment on pathologic findings and gene expression in a monocrotaline-induced pulmonary hypertension rat model. Methods: Six-week-old male Sprague-Dawley rats were allocated to 3 groups: control (C), single subcutaneous injection of normal saline (0.1 mL/kg); monocrotaline (M), single subcutaneous injection of monocrotaline (60 mg/kg); and monocrotaline + infliximab (M+I), single subcutaneous injection of monocrotaline plus single subcutaneous injection of infliximab (5 mg/kg). The rats were sacrificed after 1, 5, 7, 14, or 28 days. We examined changes in pathology and gene expression levels of TNF-${\alpha}$, endothelin-1 (ET-1), endothelin receptor A (ERA), endothelial nitric oxide synthase (eNOS), matrix metalloproteinase (MMP) 2, and tissue inhibitor of matrix metalloproteinase (TIMP). Results: The increase in medial wall thickness of the pulmonary arteriole in the M+I group was significantly lower than that in the M group on day 7 after infliximab treatment (P<0.05). The number of intraacinar muscular arteries in the M+I group was lower than that in the M group on days 14 and 28 (P<0.05). Expression levels of TNF-${\alpha}$, ET-1, ERA, and MMP2 were significantly lower in the M+I group than in the M group on day 5, whereas eNOS and TIMP expressions were late in the M group (day 28). Conclusion: Infliximab administration induced early changes in pathological findings and expression levels of TNF-${\alpha}$, and MMP2 in a monocrotaline-induced pulmonary hypertension rat model.

• 대한본초학회지
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• 제23권4호
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• pp.113-120
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• 2008

Objectives: This study was evaluated to elucidate the inhibitory potential of Imyosan(IMS) and its components, Phellodendri Cortex(PC: Phellodendron amurense Rupr., Hwangbaek in Korean) and Atractylodis Rhizoma(AR: Atratylodes lancea D.C., Changchool in Korean), on human aortic smooth muscle cells(HASMC) migration and production of matrix metalloproteinase (MMP)-2 and MMP-9 by TNF-${\alpha}$ treatment. Methods: Cytotoxic activity of IMS and its components on HASMC was using 5-(3-caroboxy meth-oxyphenyl)-2H-tetra-zolium inner salt(MTS) assay. Effect of IMS, PC and AR on TNF-${\alpha}$-induced HASMC migration underside of matrigel filter was stained with hematoxylin-eosin. And total number of cells that migrated to the underside of the filter was counted. MMP-2 and MMP-9 activity was evaluated by gelatin zymography assay. Results: The matrigel migration assay showed that IMS effectively inhibited the TNF-${\alpha}$-induced migration of HASMC. Moreover, IMS significantly inhibited MMP-9 activity. Our present study demonstrates that IMS and its components inhibit TNF-${\alpha}$-induced HASMC migration and MMP-9 activity. The inhibitory effect of IMS extract is more potent than that of its component herb extracts. Conclusions: These results provide evidence that IMS has multiple effects in the inhibition of HASMC migration and may offer a therapeutic approach to block HASMC migration.