• Nutritional Sciences
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• 제8권1호
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• pp.16-22
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• 2005

The present study examined the effects of cyclooxygenase-2 (COX-2) inhibitor celecoxib or soy-isoflavones in the presence of estrogen on apoptosis related gene expression, COX-2 and mapkinase in 48-week old female rats. Expressions of bel-2 and bax proteins, which are known to be involved in the regulation of apoptosis, were investigated in mammary glands and heart tissues. The elevated expression of bel-2 expression was observed in mammary glands of celecoxib supplemented rats as well as soy-isoflavones. The mammary glands bel-2/bax ratio was found to be higher in celecoxib or soy-isoflavones supplemented rats. However, in heart tissues, expression of bel-2 and bax was in the order of control, celecoxib and soy-isoflavones. The up-regulation of COX-2 was observed in celecoxib or soy-isoflavones in mammary glands. 'The similar trend was not displayed with the mapkinase expression. In heart tissues, the down-regulation of COX-2 as well as mapkinase was observed in celecoxib or soy-isoflavones supplemented rats. Soy-isoflavones and celecoxib both had a similar regulatory pattern of bel-2, bax and COX-2 in mammary glands, and in heart tissues, only COX-2 exhibited a similar down-regulatory properly. These findings revealed that in estrogen sufficient state, celecoxib and soy-isoflavones might not exhibit proapoptotic potential or COX-2 inhibition in normal mammary glands.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.104-104
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• 2001

Ras는 세포의 성장과 분화 등 여러 필수적인 세포기능에 없어서는 안될 중요한 역할을 담당하며 Ras가 mutation되면 암 등의 치명적인 결과를 초래한다. Ras 발현은 유방암에서 tumor aggressiveness의 지표로 간주되고 있으며 유방세포의 침습성과 연관이 있다고 알려져 있으므로 ras가 전이과정에 미치는 영향에 관한 연구는 중요한 의미를 갖는다. 본 연구의 선행연구결과, H-ras와 N-ras 모두 transformed phenotype을 나타내지만 H-ras 만이 암전이에 있어서 중요한 침윤성을 유도하는 것을 밝혔다. 이 결과는 MCF10A 세포에서 H-ras와 N-ras에 의한 신호전달경로가 각각 다른 생물학적 전이활성을 나타냄을 시사한다. 세포의 이동성은 침습성에 있어서 결정적인 역할을 하므로, 본 연구에서 H-ras와 N-ras로 형질전환된 MCF10A세포에서 이동성을 시험한 결과, 세포의 이동성이 N-ras가 아닌 H-ras MCF10A 세포에서만 크게 증가된다는 것을 보았다. 이는 침습성을 나타내는 H-ras가 세포의 이동성을 증가시키는데 작용한다는 것을 말한다. H-ras에 의해 유도된 침습성과 이동성에 대한 분자적 기전에 관하여 연구하기 위하여 H-ras MCF10A와 N-ras MCF10A 세포에서 Ras의 downstream effector들, 특히 mitogen-activated protein kinases(MAPKinases)들인 JNK1, ERK, p38의 활성화를 살펴본 결과 p38 MAPKinase가 H-ras MCF10A 세포에서 현저하게 활성화됨을 보았다. p38 MAPKinase 저해제인 SB203580를 처리하던지 dominant negative p38 (DN p38) transfectant로 p38을 불활성화시켰을 때 세포침습성 및 이동성이 저해되는 결과를 얻었다. SB203580 처리한 H-ras MCF10A 세포에서 전이에 관여하는 효소인 MMP-2 분비가 감소되었다. H-ras에 의해 유도된 침습성과 이동성은 DN JNK1 transfectant에서는 변화가 없었으나 DN MEK transfectants에서는 유의성있게 감소되었다. 이상의 결과를 종합하면, MCF10A 세포의 침윤성과 이동성에는 p38 MAPKinase 활성이 중심적인 역할을 하며, JNK 활성은 영향을 미치지 않고, ERK-1/2 활성은 충분하지는 않으나 필요하다는 것을 알 수 있었다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.107-107
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• 2001

During the last decade, enormous progress has been made on the biological significance of apoptosis. Since ras is among the most central molecule in signaling, we asked if ras regulates apoptotic pathway. We have previously shown that H-ras, but not N-ras, induces an invasiveness and motility in human breast epithelial cells (MCF10A), while both H-ras and N-ras induce transformed phenotype. In this study, we wished to seek a chemopreventive agent that effectively induces apoptosis in H-ras-activated cells. Here we show that capsaicin, the major pungent phytochemical in red pepper, induces caspase 3-involved apoptosis selectively in H-ras activated MCF10A cells while the parental MCF10A cells are not effected. In order to study the molecular mechanisms for the increased sensitivity of H-ras MCF10A cells to capsaicin-induced apoptosis, activation of ras downstream signaling molecules, mitogen-activated protein kinases (MAPKinases), upon capsaicin treatment was investigated. Phosphorylated forms of JNK1 and p38 MAPKinase were prominently increased whereas activated ERK-1/2 was decreased by capsaicin in ras-activated cells. The parental cells did not respond to capsaicin, suggesting that capsaicin selectively induces apoptosis through modulating activities of ras downstream signaling molecules in H-ras-activated cells. Studies using chemical inhibitors (CPT-cAMP, SB203580 and PD98059) and dominant negative constructs of JNKl, p38 and MEK show that activation of JNK1 and p38 MAPKinase, but not ERK-1/2, is critical for ras-mediated apoptosis by capsaicin.

• 대한수의학회지
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• 제46권4호
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• pp.315-322
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• 2006

Ethanol abuse is associated with liver injury, neurotoxicity, modulation of immune responses, and increased risk for cancer, whereas moderate ethanol consumption exerts protective effects against liver injury. However, the underlying signal transduction mechanisms of insulin-like growth factors (IGFs) which play an important regulatory role in various metabolism mechanisms are not well understood. We investigated the effects of ethanol-induced p42/44 activity on IGF-I secretion, IGF-I receptor and IGFBP-1 secretion using radioimmunoassay and western blotting in primary cultured rat hepatocytes. The p42/44 activity, IGF-I secretion and IGF-I receptor activity significantly accelerated compared to control at 10 and 30 min after 200 mM ethanol treatment, but then it became suppressed at 180 min. In contrast, IGFBP-1 secretion was inhibited compared to control at 30 min after 200 mM ethanol treatment, but increased at 180 min. The IGF-I secretion, IGF-I receptor and p42/44 activity at 30 min after 200 mM ethanol treatment accelerated with increasing ethanol concentration but IGFBP-1 secretion inhibited (p<0.05). The increased IGF-I secretion, inhibited IGFBP-1 secretion and IGF-IR activity by ethanol-induced temporal p42/44 activity at 30 min after ethanol treatment was blocked by treatment with PD98059. Alcohol dehydrogenase (ADH) inhibitor, 4-methylpyramazole blocked the changes of IGF-I secretion, IGFBP-1 secretion, and IGF-IR activity by ethanol-induced p42/44 activity at 30 and 180 min. Taken together, these results suggest that ethanol is involved in the modulation of IGF-I and IGFBP-1 secretion and IGF-IR activity by p42/44 activity in primary cultured rat hepatocytes. In addition, changing of p42/44 activity by ethanol was caused with ADH.

• 동의생리병리학회지
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• 제23권2호
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• pp.480-487
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• 2009

Atherosclerosis, slow progressing inflammatory lesion in arteries, is one of the major causes of cardiovascular diseases. As mortality due to cardiovascular disease keeps increasing in Korea, researches on pathological mechanism of atherosclerosis may be beneficial in fighting against cardiovascular diseases. It is known that migration and MMP-9 secretion of Vascular Smooth Muscle Cell(VSMC) play a significant part in pathogenesis of atherosclerosis, although detailed mechanism of entire process is not clarified. We investigated whether the seeds of Trichosanthes kirilowii maxim (TS), inhibit migration and MMP-9 production of HASMC(human aortic SMC), which were induced by TNF-${\alpha}$ treatment. Migration assay showed that TS inhibited the migration of HASMC induced by TNF-${\alpha}$, in dose dependent manner. Also by Zymography MMP-9 production of HASMC was found to be reduced by TS, both in time and in dose dependent manner. Western blotting results suggest TS suppress activity of MAPkinases.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.82-83
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• 2003

Plants have evolved differently from animals having mobile activities. Thus, plants should have developed unique defense mechanisms against biotic/abiotic stresses to which plants are differently exposed, according to seasons. Most organisms have an conserved signaling network using mitogen-activated protein kinase (MAPK) cascade(s). The phenomenon implied that they are functionally very important in all organisms. In fact, they constitute one of the major components of signaling pathways involved in regulating a wide range of cellular activities from growth and development to cell death. Recently, complete MAPK cascade was first characterized in Arabidopsis from the receptor kinase (FLS2) through fellowing MEKKI -MKK4/MKK5-MPK3/MPK6-WRKY22/MRKY29 pathway. Whereas, MAPK cascade signaling pathway in monocot plant including rice (0ryza sativa L.), the most important of all food crops and an established monocot plant research model, MAPKinase kinase kinases (MAPKKK) of rice are the first upstream component of the MAPK cascade, but MAPKKK has been first identified and characterized in our lab and designated as, OsEDRl based on its homology with the Arabidopsis EDRI. The Arabidopsis EDRl was regarded as a negative regulator of defense response and the role of rice OsEDRl was analyzed. Transcriptional regulation of OsEDRl was detected under various stresses and immunoblotting analysis is going on to detect the level of OsEDRl protein in the mutants showing unique phenotype. We also introduced the constitutively active and the dominant negative forms of the OsEDRl for characterizing biological function.

• 생명과학회지
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• 제26권1호
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• pp.34-41
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• 2016

학명이 Aronia melanocarpa인 블랙초크베리는 항산화, 항염증, 항암 효능이 뛰어난 것으로 보고되고 있다. 본 연구에서는 블랙초크베리에 대한 collagenase inhibition effects와 산화적 스트레스에 유도된 matrix metalloproteinase(MMP), MAPkinase 그리고 AP-1의 발현 그리고/또는 인산화와 같은 분자생물학적 메카니즘을 조사하였다. Collagenase inhibition 효과는 블랙초크베리 에틸아세테이트 분획물(AE)이 500 μg/ml의 농도에서 77.2% 이상의 저해효능을 나타내었고 이는 대조군인 Epigallocatechin gallate의 결과(500 μg/ml에서 83.9%)와 비교해서 유의할 만한 결과였다. Reactive oxygen species (ROS) assay는 AE에서 가장 농도의존적으로 ROS 생성이 감소되었고, 75 μg/ml의 농도에서 약 70%로 가장 낮은 활성산소가 생성되었다. MTT assay 결과, H₂O₂에 유도된 CCRF 세포에 AE를 처치하였을 때 농도의존적으로 세포 생존율이 증가하였다. 그리고 특히, AE는 H₂O₂에 유도된 CCRF 세포에서의 MMPs (MMP-1, -3 그리고 -9), MAPK (ERK, JNK 그리고 p38) 그리고 AP-1 (c-Fos와 c-Jun)의 발현과 인산화를 억제하였고, pro-collagen type I의 발현은 증가시켰다. 따라서 블랙초크베리 에틸아세테이트 분획물은 주름억제 및 콜라겐 생성의 효능이 있으며 기능성 식품 및 화장품 소재 개발 산업에서의 응용이 가능할 것으로 기대된다.

• 동의생리병리학회지
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• 제33권3호
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• pp.175-180
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• 2019

Torilis Japonica (TJ) has been used as an anti-allergy, antifungal, and antibacterial agent. Recent studies have reported that it also shows anti-cancer effects. It is report that TJ inhibits melanin synthesis in melanocyte in the skin. However, the effect and mechanism of TJ extract (TJE) on Ultraviolet (UV)B-induced photoaging are unknown. In this study, we investigated the preventive effects of TJE on matrix metalloproteinase (MMP)-1 and MMP-3 expressions and the underlying molecular mechanism in UVB-irradiated primary human dermal fibroblasts (HDFs). The effect of TJE on HDF cell viability was determined using the XTT assay and cell counting. MMP-1 and MMP-3 expressions levels were measured by western blotting and real-time PCR analysis. Activations of mitogen-activated protein kinase (MAPKinase), nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$), and activator protein-1(AP-1) were measured by western blotting. Our results showed that TJE effectively reduced UVB-induced MMP-1 and MMP-3 protein and mRNA levels. Moreover, TJE significantly blocked the UVB-induced activation of MAPK (p38 and JNK) and transcription factors ($NF-{\kappa}B$ and AP-1), but not ERK. Taken together, our results suggest that the TJE inhibits UVB-induced MMP expressions in HDFs and its may be a potential agent for the prevention and treatment of skin photoaging.