• 한국임상수의학회지
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• 제26권2호
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• pp.144-149
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• 2009

퇴행성관절염(Osteoarthritis,OA)은 관절 부위의 퇴행성 변화가 특징이며, 이를 진단하기 위해서는 연골세포나 활액에서 유래된 표지인자가 일반적으로 사용된다. 이번 연구에서는 개를 이용하여 실험적으로 퇴행성관절염을 유도하고, 활액과 연골세포에서 단백질 분해 효소인 matrix metalloproteinase(MMPs)와 MMPs의 활성을 억제시키는 것으로 알려진 tissue inhibitors of metalloproteinases(TIMPs)의 발현 정도를 조사하였다. 20마리의 비글견이 퇴행성관절염 모델로 사용되었으며 MMP-2 와 -9은 Western blot 분석에 의해서, TIMP-2의 농도는 ELISA(enzyme-linked immunosorbent assays)에 의해 결정하였다. 퇴행성관절염 유도 4주 후에 연골에서 분리한 연골세포에서 MMP-2의 발현은 증가되었지만 MMP-9의 발현은 감소되었다. 그러나, 퇴행성관절염을 유도한 개의 활액에서는 MMP-2와 -9의 발현이 모두 증가하는 것을 보였다. TIMP-2의 농도는 퇴행성관절염을 유도한 연골에서 분리한 연골세포에서는 높았지만, 활액에서는 낮은 농도를 보였다. 이러한 결과는 MMP-9가 퇴행성관절염 시 연골 조직의 변성에 따른 연골세포의 손상에 의해 MMP-2보다 더 활액으로 방출된다는 것을 보여주며, 활액 내 TIMP-2의 감소에 따른 MMPs의 활성이 퇴행성관절염을 더욱 악화시키는 것을 제안해준다. 결국 MMPs의 활성은, 특히 MMP-9, 개의 퇴행성관절염의 조기 진단과 치료를 위한 표지인자로서 사용할 수 있을 것으로 사료된다.

• Clinical and Experimental Pediatrics
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• 제53권4호
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• pp.510-518
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• 2010

목 적: 가와사끼병은 급성 전신성 미세혈관염으로서, 소아의 후천성 심질환의 가장 흔한 원인이다. 가와사끼병의 관상동맥병변에 대한 조직병리학적 소견은 미만성 혈관염에 의한 관상동맥벽의 파괴를 시사하는데, matrix metalloproteinase (MMP)와 그 내인성 억제인자인 tissue inhibitor of metalloproteinases (TIMP)가 이러한 과정에 주요한 역할을 할 것으로 생각되고 있으며, 그 중에도 MMP-9이 최근 가장 주목을 받고 있다. 따라서 본 연구에서는, 한국의 가와사끼병 환아들에서 MMP-9과 그 억제인자들인 TIMP-1, TIMP-2가 갖는 임상적 중요성을 평가하고자 하였다. 방 법: 가와사끼병 환아 47명을 연구대상으로 하였고, 급성열성 세균감염 환아 14명을 대조군으로 하였다. 혈중 MMP-9, TIMP-1, TIMP-2의 농도를 kit를 통한 효소 면역법을 이용하여 측정하였고, 각 임상 단계 및 관상동맥병변의 유무에 따라 비교분석하였다. 결 과: 급성기에 MMP-9과 TIMP-1은 가와사끼군에서 대조군에 비해 유의하게 높았고, TIMP-2는 오히려 낮았다($P$<0.05). 급성기에 증가한 MMP-9은 아급성기와 회복기에 유의하게 감소하였다($P$<0.05). 관상동맥 확장군과 비확장군의 비교에서는, 확장군에서 급성기 MMP-9과 TIMP-1, MMP-9/TIMP-2가 낮았으며, 아급성기로 가면서 통계적으로 유의하게 증가하였다가 다시 회복기로 가면서 감소하는 양상을 보였다($P$<0.05). MMP-9은 급성기와 아급성기에 TIMP-1과 통계적으로 유의한 양의 상관관계를 보였고, TIMP-2와는 아급성기와 회복기에 음의 상관관계를 보였다($P$<0.05). 결 론: 이러한 결과들은 MMP-9과 TIMP-1의 증가, 그리고 MMP-9과 TIMP-2의 불균형이 가와사끼병, 특히 그 관상동맥병변의 병태생리에 핵심적인 역할을 한다는 사실을 시사한다 하겠다. 더 종합적이고 심도 깊은 연구를 위해서 좀 더 많은 대상자를 포함하는 큰 규모의 연구들이 향후 이루어져야 할 것이다.

• Biomolecules & Therapeutics
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• 제22권5호
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• pp.414-419
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• 2014

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that regulate cell-matrix composition and are also involved in processing various bioactive molecules such as cell-surface receptors, chemokines, and cytokines. Our group recently reported that MMP-3, -8, and -9 are upregulated during microglial activation and play a role as proinflammatory mediators (Lee et al., 2010, 2014). In particular, we demonstrated that MMP-8 has tumor necrosis factor alpha (TNF-${\alpha}$)-converting enzyme (TACE) activity by cleaving the prodomain of TNF-${\alpha}$ and that inhibition of MMP-8 inhibits TACE activity. The present study was undertaken to compare the effect of MMP-8 inhibitor (M8I) with those of inhibitors of other MMPs, such as MMP-3 (NNGH) or MMP-9 (M9I), in their regulation of TNF-${\alpha}$ activity. We found that the MMP inhibitors suppressed TNF-${\alpha}$ secretion from lipopolysaccharide (LPS)-stimulated BV2 microglial cells in an order of efficacy: M8I>NNGH>M9I. In addition, MMP inhibitors suppressed the activity of recombinant TACE protein in the same efficacy order as that of TNF-${\alpha}$ inhibition (M8I>NNGH>M9I), proving a direct correlation between TACE activity and TNF-${\alpha}$ secretion. A subsequent pro-TNF-${\alpha}$ cleavage assay revealed that both MMP-3 and MMP-9 cleave a prodomain of TNF-${\alpha}$, suggesting that MMP-3 and MMP-9 also have TACE activity. However, the number and position of cleavage sites varied between MMP-3, -8, and -9. Collectively, the concurrent inhibition of MMP and TACE by NNGH, M8I, or M9I may contribute to their strong anti-inflammatory and neuroprotective effects.

• Tuberculosis and Respiratory Diseases
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• 제65권1호
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• pp.7-14
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• 2008

연구배경: 잔여 흉막비후는 결핵성 흉막염 치료 후 흔히 나타날 수 있는 합병증 중의 하나이며, 이로 인해 호흡기능에 지장을 주는 경우가 있다. 이에 결핵성 흉막염 진단 시 흉수 내의 metalloproteinase (MMP)s와 tissue inhibitor of metalloproteinase (TIMP)s의 농도가 치료 후 잔여 흉막비후가 지속되는 지 예측할 수 있는 인자가 될 수 있는 지 알아보고자 하였다. 방 법: 2004년 1월부터 2005년 6월 사이에 흉수가 발견되어 입원한 환자를 대상으로 전향적 연구를 시행하였다. 진단 시 흉수의 분석을 통해 결핵성 흉막염, 부폐렴성 흉수, 악성 흉수, 여출액군으로 나누고, 환자의 혈청과 흉수에서 ELISA 방법을 이용하여 MMP-1, -2, -8, -9와 TIMP-1, -2를 측정하였다. 결핵성 흉막염의 경우 흉부엑스선검사로 항결핵제 치료 종결 시점과 마지막 추적 관찰시점에 잔여 흉막비후의 두께를 측정하여 잔여 흉막비후가 있는 군과 없는 군으로 나누었다. 결 과: 흉수가 발견되어 입원한 환자 중 제외 기준에 해당하는 환자를 제외하고 총 39명의 환자가 대상이 되었다. 이 중 결핵성 흉막염은 23명, 부폐렴성 흉수 7명, 악성 흉수 7명, 여출액 2명이었다. 결핵성 흉막염 환자 23명 중 본원에서 항결핵제 치료를 종료한 환자는 17명이었으며, 이 중 잔여 흉막비후가 없는 군은 10명(59%)이었으며, 잔여 흉막비후가 있는 군은 7명(41%)이었다. 잔여 흉막비후가 있는 군은 흉수 TIMP-1 ($41,405.9{\pm}9,737.3ng/mL$)이 잔여 흉막비후가 없는 군($29,134.9{\pm}8,801.8$)보다 의미있게 높았다(p=0.032). 치료 종료 후 평균 $8{\pm}5$개월의 추적관찰이 가능한 13명의 환자들에서, 마지막으로 촬영한 흉부 후전위 촬영에서 잔여 흉막비후가 없는 군은 11명(85%)이었고, 잔여 흉막비후가 있는 군은 2명(15%)이었다. 잔여 흉막비후가 있는 군은 흉수 TIMP-2 ($34.4{\pm}6.5ng/mL$)가 잔여 흉막비후가 없는 군($44.4{\pm}15.5$)보다 의미있게 낮았다(p=0.038). 결 론: 결핵성 흉막염의 잔여 흉막비후의 발생에 TIMP-1과 TIMP-2이 관여 될 수도 있을 것으로 추정된다.

• 약학회지
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• 제48권3호
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• pp.189-196
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• 2004

Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) playa key role in tumor invasion and metastasis. As an inhibitor of MMP-2, TIMP-2 is known to block both the invasive and metastatic behavior of cancer cells, and decrease tumor growth activity. We performed this study to investigate the effects of TIMP-2 over-expression induced by retroviral mediated gene transfer in vitro and in vivo. The human colon cancer cell line SW480 was transfected with the retroviral vector encoding TIMP-2. The effects of TIMP-2 over-expression were analyzed by invasion assay and gelatinase activity test in colon cancer cells and tumorigencity in nude mice. In evaluation of the transfection efficiency of the retroviral vector encoding TIMP-2 in colon cancer cells, we confirmed up-regulation of TIMP-2 expression dependent on the time of cell culture. In addition, inhibition of MMP-2 expression in SW480/TIMP-2 was shown by gelatin zymography. In the in vitro invasion assay SW480/TIMP-2 inhibited the invasiveness on matrigel coated with collagen. To determine whether TIMP-2 can modulate in vivo tumorigenicity and metastasis, SW480/TIMP-2 cells were injected subcutaneously in nude mice. The tumor mass formation of SW480/TIMP-2 cells in nude mice was markedly decreased compared to nontransfected cancer cells. These results showed that colon cancer cells transfected with the retroviral vector encoding TIMP-2 inhibits the invasiveness in vitro and tumorigenicity in vivo.

• The Korean Journal of Physiology and Pharmacology
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• 제14권4호
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• pp.199-204
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• 2010

The potential therapeutic action of shikonin in an experimental model of rheumatoid arthritis (RA) was investigated. As a RA animal model, DBA/1J mice were immunized two times with type II collagen. After the second collagen immunization, mice were orally administered shikonin (2 mg/kg) once a day for 35 days, and the incidence, clinical score, bone mineral density (BMD), bone mineral content (BMC) and joint histopathology were evaluated. BMD in the proximal regions of the tibia largely increased in the shikonin treatment group compared with the control group. We also examined the effect of shikonin on inflammatory cytokines and cartilage protection. Shikonin treatment significantly reduced the incidence and severity of collagen-induced arthritis (CIA), markedly abrogating joint swelling and cartilage destruction. Shikonin also significantly inhibited the production of matrix metalloproteinase (MMP)-1 and up-regulated tissue inhibitors of metalloproteinase (TIMP)-1 in mice with CIA. In conclusion, shikonin exerted therapeutic effects through regulation of MMP/TIMP; these results suggest that shikonin is an outstanding candidate as a cartilage protective medicine for RA.

• Journal of Ginseng Research
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• 제36권1호
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• pp.61-67
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• 2012

In the present study, effects of sun ginseng (SG) on the collagen synthesis and the proliferation of dermal fibroblast were investigated. Collagen synthesis was measured by assaying procollagen type I C-peptide production. In addition, the level of matrix metalloproteinase (MMP)-1 was assessed by western blot analysis. SG suppressed the MMP-1 protein level in a dose-dependent manner. In contrast, SG dose-dependently increased tissue inhibitors of MMP (TIMP)-1 production in fibroblasts. SG increased type I collagen production directly and/or indirectly by reducing MMP-1 and stimulating TIMP-1 production in human dermal fibroblasts. SG dose-dependently induced fibroblast proliferation and this, in turn, can trigger more collagen production. These results suggest that SG may be a potential pharmacological agent with anti-aging properties in cultured human skin fibroblast.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.379-384
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• 2004

Fibroblast growth factor-2 (FGF-2) is known to modulate numerous cellular functions in various cell types, including cell proliferation, differentiation, survival, adhesion, migration, and motility, and also in processes such as wound healing, angiogenesis, and vasculogenesis. FGF-2 regulates the expression of several molecules thought to mediate critical steps during angiogenesis. This study examines the mechanisms underlying FGF-2-induced cell migration, using human umbilical vein endothelial cells (HUVECs). FGF-2 induced the nondirectional and directional migration of endothelial cells, which are inhibited by MMPs and plasmin inhibitors, and induced the secretion of matrix metalloproteinase-3 (MMP3) and MMP-9, but not MMP-l and MMP-2. FGF-2 also induced the secretion of the tissue inhibitor of metalloproteinase-l (TIMP-I), but not of TIMP- 2. Also, the pan-PKC inhibitor inhibited FGF-2-induced MMP-9 secretion. It is, therefore, suggested that FGF-2 induces the migration of cultured endothelial cells by means of increased MMPs and plasmin secretion. Furthermore, FGF-2 may increase MMP-9 secretion by activating the PKC pathway.

• Molecules and Cells
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• 제25권1호
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• pp.105-111
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• 2008

Radiotherapy is an important treatment for many malignant tumors, but there are recent reports that radiation may increase the malignancy of cancer cells by stimulating expression of type IV collagenases. In this study, we examined changes in matrix metalloproteinase (MMP) inhibitors, such as the tissue inhibitors of metalloproteinase (TIMP)-1, TIMP-2 and RECK, in response to irradiation in Panc-1 pancreatic cancer cells. Irradiation increased RECK protein levels but not mRNA levels, whereas no significant changes were found in TIMP-1 and TIMP-2. The enhanced RECK protein levels were associated with an increase in MMP inhibitory activity. However, irradiation slightly but reproducibly increased the invasiveness of the Panc-1 cells. Like irradiation, treatment of Panc-1 cells with transforming growth factor $(TGF)-{\beta}1$ led to a 2-fold increase in RECK protein levels. Transient transfection with Smad3 also increased RECK protein levels, but transfection with Smad7 markedly reduced them. Stable expression of Smad7 and treatment with SB431542, an inhibitor of $TGF-{\beta}$ receptor I kinase, abolished $TGF-{\beta}1$- and radiation-mediated effects on RECK. Furthermore, irradiation increased levels of phosphorylated Smad3. We conclude that radiation post-transciptionally enhances RECK protein levels in Panc-1 cells, at least in part, via $TGF-{\beta}$ signaling, and that irradiation increases Panc-1 invasiveness via a mechanism that may not be linked to MMP-2 activity.

• Asian Pacific Journal of Cancer Prevention
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• 제15권14호
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• pp.5747-5751
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• 2014

Background: Coptisine, an isoquinoline alkaloid extracted from Coptidis rhizoma, has many biological activities such as antidiabetic, antimicrobial and antiviral actions. However, whether coptisine exerts anti-cancer metastasis effects remains unknown. Materials and Methods: Effects of coptisine on highly metastatic human breast cancer cell MDA-MB-231 proliferation were evaluated by trypan blue assay and on cell adhesion, migration and invasion by gelatin adhesion, wound-healing and matrigel invasion chamber assays, respectively. Expression of two matrix metalloproteinases (MMPs), MMP-9, MMP-2 and their specific inhibitors tissue inhibitor of metalloproteinase 1 (TIMP-1) and tissue inhibitor of metalloproteinase 2 (TIMP-2) were analyzed by RT-PCR. Results: Coptisine obviously inhibited adhesion to an ECM-coated substrate, wound healing migration, and invasion through the matrigel in MDA-MB-231 breast cancer cells. RT-PCR revealed that coptisine reduced the expression of the ECM degradation-associated gene MMP-9 at the mRNA level, and the expression of TIMP-1 was upregulated in MDA-MB-231 cells, while the expression of MMP-2 and its specific inhibitor TIMP-2 was not affected. Conclusions: Taken together, our data showed that coptisine suppressed adhesion, migration and invasion of MDA-MB-231 breast cancer cells in vitro, the down-regulation of MMP-9 in combination with the increase of TIMP-1 possibly contributing to the anti-metastatic function. Coptisine might be a potential drug candidate for breast cancer therapy.