• Molecules and Cells
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• 제44권1호
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• pp.13-25
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• 2021

Apoptosis and compensatory proliferation, two intertwined cellular processes essential for both development and adult homeostasis, are often initiated by the mis-regulation of centrosomal proteins, damaged DNA, and defects in mitosis. Fly Anastral spindle 3 (Ana3) is a member of the pericentriolar matrix proteins and known as a key component of centriolar cohesion and basal body formation. We report here that ana3m19 is a suppressor of lethality induced by the overexpression of Sol narae (Sona), a metalloprotease in a disintegrin and metalloprotease with thrombospondin motif (ADAMTS) family. ana3m19 has a nonsense mutation that truncates the highly conserved carboxyl terminal region containing multiple Armadillo repeats. Lethality induced by Sona overexpression was completely rescued by knockdown of Ana3, and the small and malformed wing and hinge phenotype induced by the knockdown of Ana3 was also normalized by Sona overexpression, establishing a mutually positive genetic interaction between ana3 and sona. p35 inhibited apoptosis and rescued the small wing and hinge phenotype induced by knockdown of ana3. Furthermore, overexpression of Ana3 increased the survival rate of irradiated flies and reduced the number of dying cells, demonstrating that Ana3 actively promotes cell survival. Knockdown of Ana3 decreased the levels of both intra- and extracellular Sona in wing discs, while overexpression of Ana3 in S2 cells dramatically increased the levels of both cytoplasmic and exosomal Sona due to the stabilization of Sona in the lysosomal degradation pathway. We propose that one of the main functions of Ana3 is to stabilize Sona for cell survival and proliferation.

• Nutrition Research and Practice
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• 제5권5호
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• pp.375-380
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• 2011

This study investigated the effects of inorganic sulfur on metastasis in MDA-MB-231 human breast cancer cells. MDA-MB-231 cells were cultured in the absence or presence of various concentrations (12.5, 25, or 50 ${\mu}mol$/L) of inorganic sulfur. Cell motility, invasion, and the activity and mRNA expression of matrix metalloproteases (MMPs) were examined. Numbers of viable MDA-MB-231 cells did not differ by inorganic sulfur treatment from 0 to 50 ${\mu}mol$/L within 48 h. Inorganic sulfur significantly decreased cell motility and invasion in the MDA-MB-231 cells in a dose-dependent manner (P<0.05), as determined using a Boyden chamber assay and a Matrigel chamber. The activities of MMP-2 and MMP-9 were significantly reduced by inorganic sulfur in a dose-dependent manner (P<0.05). The inorganic sulfur also significantly inhibited MMP-2 and MMP-9 expression in the cells (P<0.05). These data suggest that inorganic sulfur can suppress cancer cell motility and invasion by inhibiting MMP-2 and MMP-9 activity and gene expression in MDA-MB-231 cells.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.168.3-169
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• 2003

The matrix metalloproteases (MMPs) play important roles in invasion, metastasis and angiogenesis in various cell types. Tissue inhibitor of metalloprotease (TIMP)-2, an endogenopus inhibitor of MMP-2, has been shown to inhibit invasion and metastasis. We have previously shown that MMP-2 is responsible for the H-ras-induced invasive and migrative phenotypes in MCF10A human breast epithelial cells. Here, we investigated the effect of TlMP-2 overexpression on invasion and migration in H-ras MCF10A cells. (omitted)

• 대한화장품학회지
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• 제30권2호
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• pp.241-246
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• 2004

발아 과정을 통한 식물 종자의 변화를 확인하고, 발아된 식물종자로부터 새로운 펩타이드를 개발하여 항노화 화장품 소재로서의 가능성을 확인하였다. 발아과정을 통한 식물 종자의 변화는 단백질 양 측정, 겔 투과 크로마토그램(GPC)과 SDS-PAGE를 통한 단백질 분자량 분포 변화, 아미노산 조성분석을 통해 발아 전과 비교하였다. 발아된 검은 쌀로부터 펩타이드를 생산하기 위해 단백질 분해효소를 처리하였으며, 생산된 발아 검은 쌀 펩타이드의 효과를 발아 검은 쌀 단백질과 비교하여 확인하였다. In vitro matrix-metalloprotease (MMP) 활성 저해효과는 형광 정량법을 이용하였으며, 인간 섬유아세포 활성화 효과를 확인하였고, 콜라겐 생합성 촉진효과와 자외선 조사에 의한 MMP 발현 저해효과는 효소면역분석법(ELISA)을 이용하여 확인하였다. 발아에 의해 검은 쌀의 단백질량, 단백질 분자량 분포 및 아미노산의 조성이 변화함을 확인하였으며, 검은 쌀의 발아와 단백질 분해효소중 파파인 처리를 병행함으로서, 상대적으로 낮은 분자량을 갖는 신규 펩타이드의 제조가 가능하였다. GPC를 이용하여 제조된 발아 검은 쌀 펩타이드의 분자량 분포를 확인한 결과, 대부분이 900dalton 이하의 분자량을 가지는 것을 확인하였으며, 또한, 이러한 발아 검은 쌀 펩다이드는 약 40% 정도의 인간 섬유아세포 활성화 효과를 가지며, 또한 농도 의존적으로 콜라겐 분해효소 활성 저해효과를 가지는 것을 확인하였다. 또한, 자외선 조사에 의해 유도되는 콜라겐 분해효소 발현을 약 50% 억제하며, 콜라겐 생합성을 약 20% 촉진하는 것을 확인할 수 있었다.

• 생명과학회지
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• 제29권11호
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• pp.1192-1199
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• 2019

본 연구는 화장품 소재로서 우절 에탄올추출물의 가능성을 확인하기 위한 것이다. 이를 위해 우리는 연근의 마디부분을 70%에탄올로 추출하여 사용하였으며 미백효능 및 주름개선 효과에 대한 생물학적 활성 평가를 수행하였다. 시료의 미백활성을 알아보기 위해 멜라닌세포(B16F10 cells)의 MTT assay를 이용한 샘플의 독성평가와 멜라닌 생성에 영향을 주는 관련 단백질의 발현량을 확인하였다. 시료의 주름활성억제 평가를 위해 collagenase inhibition assay를 수행하였다. 또한 UVB로 유도된 섬유아세포(CCD-986sk cells)의 MTT assay를 이용한 샘플의 독성평가와 matrix metalloprotease (MMP-1) inhibition 및 procollagen synthesis를 평가하였다. 미백활성 평가를 알아보기 위한 western bolt 결과, 멜라닌 생성과 관련된 두 단백질 Tyrosinase related protein-1 (TRP-1, Tyrosinase related protein-2 TRP-2)의 합성이 농도의존적으로 감소됨을 나타내었다. 게다가 우절 에탄올추출물의 collagenase inhibiion 실험의 결과, $500{\mu}g/ml$의 농도에서 대조군인 EGCG와 효능이 비슷한 80% 이상의 효과를 나타내었다. Procollagen synthesis 결과 UVB 자극군에 의해 콜라겐 합성은 68.8%로 감소되었으며 시료의 $25{\mu}g/ml$의 농도에서 80.2%의 콜라겐 합성에 회복능을 보였다. MMP-1 inhibition 실험결과는 $25{\mu}g/ml$ 농도에서 20.2%의 저해능을 나타내었다. 실험 결과는 NRN의 미백 및 주름 억제 효과를 검증하고, 향후 안전한 천연 화장품 재료로 사용될 수 있음을 확인했다.

• 대한의생명과학회지
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• 제19권1호
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• pp.9-16
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• 2013

Elevated blood triglyceride (TG) levels correlate with development of atherosclerosis suggesting that TG may promote the development of this disease. During atherosclerosis, TG is taken up by tissue macrophages which result in dramatic changes in various secreted factors. One such factor is the family of matrix metalloproteases (MMP) which are involved in tissue remodeling during both physiological and pathological processes. In this study, we examined the MMP expression profile in PMA-differentiated THP-1 macrophages treated with TG. We found that TG-treated THP-1 macrophages showed decreased expression of MMP-3, MMP-7, MMP-8 and MMP-9 in a time- and dose-dependent manner. In contrast, expression of MMP-1, MMP-2, and MMP-10 remained relatively unchanged after TG treatment. In addition, we found that expression of select MMPs was affected by various inhibitors of signaling pathways. In particular, expression of MMP-3 was slightly recovered by cRAF and PLC signaling pathway inhibitors. These data suggests a possible role of MMPs in macrophages during TG-induced atherosclerosis.

• 생명과학회지
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• 제22권12호
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• pp.1600-1607
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• 2012

Matrix metalloproteinase (MMP)는 세포외골격의 주요 조절효소로, 배아발생, 혈관생성, 상처치료 및 조직 재생과정에 중요한 인자로 알려져 있다. MMP의 조절 이상은 비정상적 세포외골격 분해로 인해 암 전이와 같은 질병을 일으킨다. 따라서, MMP의 발현과 활성은 엄격하게 조절되고 있다. 최근, 초파리 Mmp1이 소화기관에서 강하게 발현되며, 장줄기세포의 비정상적인 활성을 억제하여 장의 항상성 유지에 중요함을 밝혔다. 하지만, 장조직에서 Mmp1의 발현 조절 기전은 아직 밝혀지지 않았다. 본 연구에서는, 장조직에서 Mmp1의 발현이 장 발생과 항상성 유지에 중요한 Caudal homeobox 유전자에 의해 조절되는지를 연구하였다. GAL4/UAS 조절계를 이용하여 장조직 특이적으로 Caudal의 발현을 감소시켰을 때, Mmp1의 발현이 감소함을 확인하였으며, Caudal을 과발현 시켰을 때, Mmp1의 발현이 증가함을 in vitro와 in vivo 실험 모두에서 확인하였다. 또한, Mmp1 promoter에 Caudal 전사인자 결합 부위가 존재하며, 이 부위가 Mmp1 발현에 중요한 역할을 함을 확인하였다. 이상의 본 연구는, 정상적 혹은 암화 과정에서 Mmp1이 Caudal의 표적 유전자일 수 있음을 의미한다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.90-90
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• 2018

Kochiae Fructus is a medicinal plant used as medicine of the urinary organs, skin disease and inflammation. In this study, anti-inflammatory activity of Kochiae Fructus extract and its possible mechanisms of action were examined. The anti-inflammatory activity was investigated by inhibition of lipopolysaccharide (LPS) induced nitric oxide (NO), pro-inflammatory cytokine production and matrix metalloprotease-9 (MMP-9) in murine macrophage-like cell line Raw 264.7 cells. The measurement of the induced pro-inflammatory cytokine levels were carried out by ELISA. The phosphorylation of ERK1/2, JNK and MAPK and the nuclear expression of nuclear factor $NF-{\kappa}B$ p65 were investigated by Wesern blot analysis. The extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of $NF-{\kappa}B$ p65 in activated cells. As a result, we suggest that the extract of Kochiae Fructus decreased the production of pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6), nitric oxide, MMP-9 in LPS-induced Raw264.7 cells. Kochiae Fructus has possibility to be used as therapeutic benefits against inflammatory diseases.

• Journal of Korean Neurosurgical Society
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• 제53권6호
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• pp.323-330
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• 2013

Objective : To develop a simple, reproducible model of disc degeneration in rabbits through percutaneous annular puncture and to confirm the degree of degeneration over time. Methods : Fifteen New Zealand white rabbits (4 to 5 months old and weighing approximately 3 to 3.5 kg each) underwent annular puncture of the L2-L3, L3-L4, and L4-L5 discs. Rabbits were sacrificed at 4, 8, or 20 weeks after puncture. For a longitudinal study to assess changes in disc height over time, serial X-rays were performed at 0, 2, 4, 8, and 20 weeks for rabbits in the 20-week group. Upon sacrifice, the whole spinal column and discs were extracted and analyzed with magnetic resonance imaging (MRI), real time reverse transcriptase-polymerase chain reaction, and histological staining. Results : The X-rays showed a slow, progressive decrease in disc height over time. Significant disc space narrowing compared to preoperative disc height was observed during the time period (p<0.001). The MRI grade, aggrecan, and matrix metalloprotease-13 mRNA expression and hematoxylin and eosin/safranin O/anti-collagen II staining were consistently indicative of degeneration, supporting the results of the X-ray data. Conclusion : Percutaneous annular puncture resulted in slow, reproducible disc degeneration that was confirmed by radiology, biochemistry, and histology. This in vivo model can be used to study and evaluate the safety and efficacy of biologic treatments for degenerative disc disease.

• BMB Reports
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• 제56권8호
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• pp.439-444
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• 2023

Emphysema is a chronic obstructive lung disease characterized by inflammation and enlargement of the air spaces. Regorafenib, a potential senomorphic drug, exhibited a therapeutic effect in porcine pancreatic elastase (PPE)-induced emphysema in mice. In the current study we examined the preventive role of regorafenib in development of emphysema. Lung function tests and morphometry showed that oral administration of regorafenib (5 mg/kg/day) for seven days after instillation of PPE resulted in attenuation of emphysema. Mechanistically, regorafenib reduced the recruitment of inflammatory cells, particularly macrophages and neutrophils, in bronchoalveolar lavage fluid. In agreement with these findings, measurements using a cytokine array and ELISA showed that expression of inflammatory mediators including interleukin (IL)-1β, IL-6, and CXCL1/KC, and tissue inhibitor of matrix metalloprotease-1 (TIMP-1), was downregulated. The results of immunohistochemical analysis confirmed that expression of IL-6, CXCL1/KC, and TIMP-1 was reduced in the lung parenchyma. Collectively, the results support the preventive role of regorafenib in development of emphysema in mice and provide mechanistic insights into prevention strategies.