• The Korean Journal of Physiology and Pharmacology
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• 제9권6호
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• pp.315-322
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• 2005

In this study, the authors investigated whether death of vascular smooth muscle cell (VSMC) had a pathological pertinence. Conditioned media obtained from rat aorta smooth muscle cell (SMC) that were induced death by expressing FADD in the absence of tetracycline (FADD-SMC) triggered death of normal SMC. DNA fragmentation and caspase-3 activation were observed in dying SMC by conditioned media. FADD-SMC showed transcriptional activation of tumor necrosis factor $(TNF)-{\alpha}$. Conditioned medium contained $TNF-{\alpha}$, indicating secretion of the cytokine from dying FADD-SMC. It was investigated if secreted $TNF-{\alpha}$ was functional. Conditioned medium activated ERK and p38 MAPK pathways and induced MMP-9 expression, whereas depletion of the cytokine with its soluble receptor (sTNFR) remarkably inhibited induction of MMP-9 by conditioned medium. These findings suggest that $TNF-{\alpha}$ in conditioned medium seems to be active. Then, contribution of $TNF-{\alpha}$ on death-inducing activity of conditioned medium was examined. Depletion of $TNF-{\alpha}$ with soluble $TNF-{\alpha}$ receptor decreased the death activity of conditioned medium by 35%, suggesting that $TNF-{\alpha}$ play a partial role in the death activity. Boiling of medium almost completely abolished the death-inducing activity, suggesting that other heat labile death inducing proteins existed in conditioned medium. Taken together, these results indicate that SMC undergoing death could contribute to inflammation by expressing inflammatory cytokines and pathological complications by inducing death of neighboring cells.

• 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.

• The Korean Journal of Physiology and Pharmacology
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• 제24권3호
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• pp.241-248
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• 2020

Luminespib (AUY922), a heat shock proteins 90 inhibitor, has anti-neoplastic and antitumor effects. However, it is not clear whether AUY922 affects events in vascular diseases. We investigated the effects of AUY922 on the platelet-derived growth factor (PDGF)-BB-stimulated proliferation and migration of vascular smooth muscle cells (VSMC). VSMC viability was detected using the XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) reagent. To detect the attenuating effects of AUY922 on PDGF-BB-induced VSMCs migration in vitro, we performed the Boyden chamber and scratch wound healing assays. To identify AUY922-mediated changes in the signaling pathway, the phosphorylation of protein kinase B (Akt) and extracellular signal-regulated kinase (ERK) 1/2 was analyzed by immunoblotting. The inhibitory effects of AUY922 on migration and proliferation ex vivo were tested using an aortic ring assay. AUY922 was not cytotoxic at concentrations up to 5 nM. PDGF-BB-induced VSMC proliferation, migration, and sprout outgrowth were significantly decreased by AUY922 in a dose-dependent manner. AUY922 significantly reduced the PDGF-BB-stimulated phosphorylation of Akt and ERK1/2. Furthermore, PD98059 (a selective ERK1/2 inhibitor) and LY294002 (a selective Akt inhibitor) decreased VSMC migration and proliferation by inhibiting phosphorylation of Akt and ERK1/2. Greater attenuation of PDGF-BB-induced cell viability and migration was observed upon treatment with PD98059 or LY294002 in combination with AUY922. AUY922 showed anti-proliferation and anti-migration effects towards PDGF-BB-induced VSMCs by regulating the phosphorylation of ERK1/2 and Akt. Thus, AUY922 is a candidate for the treatment of atherosclerosis and restenosis.

• The Korean Journal of Physiology and Pharmacology
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• 제18권1호
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• pp.15-23
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• 2014

Transient receptor potential melastatin 7 (TRPM7) is a member of the melastatin-related subfamily and contains a channel and a kinase domain. TRPM7 is known to be associated with cell proliferation, survival, and development. It is ubiquitously expressed, highly permeable to $Mg^{2+}$ and $Ca^{2+}$, and its channel activity is negatively regulated by free $Mg^{2+}$ and Mg-complexed nucleotides. Recent studies have investigated the relationships between TRPM7 and a number of diseases. TRPM7 regulates cell proliferation in several cancers, and is associated with ischemic cell death and vascular smooth muscle cell (VSMC) function. This review discusses the physiologic and pathophysiologic functions and significance of TRPM7 in several diseases.

• Biomolecules & Therapeutics
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• 제19권2호
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• pp.187-194
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• 2011

Atherosclerosis and post-angiography restenosis are associated with intimal thickening and concomitant vascular smooth muscle cell (VSMC) proliferation. Obovatol, a major biphenolic component isolated from the Magnolia obovata leaf, is known to have anti-inflammatory and anti-tumor activities. The goal of the present study was to enhance the inhibitory effects of obovatol to improve its potential as a preventive or therapeutic agent in atherosclerosis and restenosis. Platelet-derived growth factor (PDGF)-BB-induced proliferation of rat aortic smooth muscle cells (RASMCs) was examined in the presence or absence of a newly synthesized obovatol derivative, OD78. The observed anti-proliferative effect of OD78 was further investigated by cell counting and [$^3H$]-thymidine incorporation assays. Treatment with 1-4 ${\mu}M$ OD78 dose-dependently inhibited the proliferation and DNA synthesis of 25 ng/ml PDGF-BB-stimulated RASMCs. Accordingly, OD78 blocked PDGF-BB-induced progression from the $G_0/G_1$ to S phase of the cell cycle in synchronized cells. OD78 decreased the expression levels of CDK4, cyclin E, and cyclin D1 proteins, as well as the phosphorylation of retinoblastoma protein and proliferating cell nuclear antigen; however, it did not change the CDK2 expression level. In addition, OD78 inhibited downregulation of the cyclin-dependent kinase inhibitor (CKI) $p27^{kip1}$. However, OD78 did not affect the CKI $p21^{cip1}$ or phosphorylation of early PDGF signaling pathway. These results suggest that OD78 may inhibit PDGF-BB-induced RASMC proliferation by perturbing cell cycle progression, potentially through $p27^{kip1}$ pathway activation. Consequently, OD78 may be developed as a potential anti-proliferative agent for the treatment of atherosclerosis and angioplasty restenosis.

• The Korean Journal of Physiology and Pharmacology
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• 제17권3호
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• pp.203-208
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• 2013

As the abnormal proliferation of vascular smooth muscle cells (VSMCs) plays a critical role in the development of atherosclerosis and vascular restenosis, a candidate drug with antiproliferative properties is needed. We investigated the antiproliferative action and underlying mechanism of a newly synthesized naphthoquinone derivative, 5,8-dimethoxy-2-nonylamino-naphthalene-1,4-dione (2-nonylamino-DMNQ), using VSMCs treated with platelet-derived growth factor (PDGF). 2-Nonylamino-DMNQ inhibited proliferation and cell number of VSMCs induced by PDGF, but not epidermal growth factor (EGF), in a concentration-dependent manner without any cytotoxicity. This derivative suppressed PDGF-induced $[^3H]$-thymidine incorporation, cell cycle progression from $G_0/G_1$ to S phase, and the phosphorylation of phosphor-retinoblastoma protein (pRb) as well as the expression of cyclin E/D, cyclin-dependent kinase (CDK) 2/4, and proliferating cell nuclear antigen (PCNA). Importantly, 2-nonylamino-DMNQ inhibited the phosphorylation of PDGF receptor${\beta}$(PDGF-$R{\beta}$) enhanced by PDGF at $Tyr^{579}$, $Tyr^{716}$, $Tyr^{751}$, and $Tyr^{1021}$ residues. Subsequently, 2-nonylamino-DMNQ inhibited PDGF-induced phosphorylation of STAT3, ERK1/2, Akt, and $PLC{\gamma}1$. Therefore, our results indicate that 2-nonylamino-DMNQ inhibits PDGF-induced VSMC proliferation by blocking PDGF-$R{\beta}$ autophosphorylation, and subsequently PDGF-$R{\beta}$-mediated downstream signaling pathways.

• Journal of Chest Surgery
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• 제40권4호
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• pp.264-272
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• 2007

배경: 혈관내피 성장인자(vascular endothelial growth factor, VEGF)는 혈관평활근세포(vascular smooth muscle cell)의 증식과 이동을 촉진함으로써 혈관신생에 중요한 역할을 한다. 당뇨병은 VEGF의 발현과 연관되어 정상 혈당상태에서 보다 세포의 증식을 더욱 촉진시킨다. 당뇨병쥐에서 VEGF 수용체의 선택적 차단이 손상된 혈관에서 신내막 형성과 혈관평활근세포의 이동에 미치는 영향에 대해 알아보고자 했다. 대상 뜻 방법: 당뇨병 쥐의 경동맥 풍선손상 모델에서 위약을 투여하거나, 혈관내피 성장 인자 수용체-1(VEGFR-1)에 선택적으로 작용하는 항-Flt-1 펩타이드(anti-Flt-1 peptide; Gly-Asn-Gln-Trp-Phe-Ile)를 풍선손상 2일 전부터 0.5mg/kg의 용량으로 2주간 매일 투여한 군으로 나누어 Hematoxylin-Eosin 염색을 하여 신내막의 형성정도와 혈관내강의 협착정도를 비교하였으며, proliferative cell nuclear antigen (PCNA)에 대한 면역조직화학염색법을 시행하여 세포의 증식정도를 관찰하였다. 혈관평활근세포를 고혈당환경에서 배양하고 transwell assay를 시행하여 혈관평활근세포의 이동 정도를 측정하였다. 고혈당 환경에서 자라고 있는 혈관평활근세포에 50ng/mL의 VEGF를 단독 또는 3ug/mL의 항-Flt-1 펩타이드와 함께 처리하고 일정시간이 지난 후 matrigel filter를 통과한 세포를 세어 아무런 처치를 받지 않은 세포가 이동한 정도와 비교하였다. 또한, 혈관평활근세포에 세포이동 정도 측정 시와 같은 처리를 한 후, RNA를 분리하고 reverse transcription-polymerase chain reaction (RT-PCR)을 시행하여 기질금속단백분해효소(matrix metalloprotenase, MMP)의 발현 정도를 관찰하였다. 결과: 신내막의 면적은 위약 투여 쥐는 $0.24{\pm}0.03 mm^2$이었으나, 항-Flt-1 펩타이드의 처리에 의해 $0.15{\pm}0.04 mm^2$로 유의하게 감소하였으며(p<0.01), 신내막 형성에 따른 내강의 협착 정도도 위약 투여 쥐는 $61.85{\pm}5.11%$, 항-Flt-1 펩타이드 투여 쥐는 $36.03{\pm}3.78%$로 항-Flt-1 펩타이드 투여에 의하여 유의하게 감소하였다(p<0.01). 신내막의 전체 세포수에 대한 PCNA(+)인 세포를 백분율로 구하였으며, 위약 투여 쥐와 항- Flt-1 펩타이드 투여 쥐에서 각각 $52.82{\pm}4.20%,\;38.11{\pm}6.89%$로 나타나 항-Flt-1 펩타이드를 투여한 쥐에서 PCNA(+)인 세포가 유의하게 적음을 보이고 있다(p<0.05). 혈관평활근세포의 이동 정도 측정에서는 항-Flt-1 펩타이드 처리에 의하여 VEGF에 의한 혈관평활근 세포의 이동이 유의하게 감소하였다(p<0.01). 또한, 항-Flt-1 펩타이드 처리에 의하여 VEGF에 의한 MMP-3와 MMP-9 mRNA의 발현 증가가 억제되었다. 결론: 항-Flt-1 펩타이드는 당뇨병쥐의 경동맥손상모델에서 신내막 형성을 억제하였으며, 고혈당 환경에서 배양된 혈관평활근세포의 이동과, MMP-3와 MMP-9의 활성을 억제하였다.

• Food Science and Biotechnology
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• 제17권2호
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• pp.247-250
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• 2008

The $CHCl_3$, EtOAc, and n-BuOH fractions showed a marked inhibition of 5% fatal bovine serum (FBS)-induced cell proliferation. The $IC_{50}$ values of the chloroform fractions from leaf, stem, and root as well as the n-BuOH and EtOAc fraction from root on cell proliferation were $1.2{\pm}0.4$, $17.2{\pm}6.4$, $81.8{\pm}33.2$, $40.8{\pm}8.0$, and $237.1{\pm}85.6\;{\mu}g/mL$, respectively. On the other hand, the EtOAc fractions, and the $CHCl_3$ fraction significantly inhibited collagen-, arachidonic acid-, U46619-, and thrombin-induced platelet aggregations. The $IC_{50}$ values of EtOAc fraction from leaf, and the $CHCl_3$ and EtOAc fraction from stem were $214.1{\pm}12.2$, $134.3{\pm}2.5$, and $42.6{\pm}7.0\;{\mu}g/mL$ with collagen, $312.4{\pm}7.5$, $158.9{\pm}1.7$, and $82.2{\pm}2.7\;{\mu}g/mL$ with arachidonic acid, $31.1{\pm}2.4$, $48.7{\pm}0.3$, and $29.7{\pm}1.1\;{\mu}g/mL$ with U46619, and $36.7{\pm}2.4$, $69.1{\pm}11.3$, and $34.2{\pm}0.1\;{\mu}g/mL$ with thrombin, respectively. Taken together, these data provide new evidence that fractions from Allium victorialis var. platyphyllum (AVP) are able to inhibit vascular smooth muscle cell (VSMC) proliferation and platelet aggregation, which may be a novel resource for the development of anti-atherothrombotic agents.

• 생명과학회지
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• 제28권12호
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• pp.1516-1522
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• 2018

죽상동맥경화는 대동맥의 만성염증에 의해 주로 발병되는 폐쇄동맥질환이다. 혈관평활근세포의 증식 및 이동은 죽상동맥경화 발병의 주된 병리적 반응이다. 본 연구에서는 죽상동맥경화 발병기전을 유도하는 표적 염증반응 물질의 탐색 및 이들에 의한 신호전달 기전을 연구하였다. 혈관평활근세포의 증식 및 이동은 prostaglandin $F_{2{\alpha}}$ ($PGF_{2{\alpha}}$)에 의해 의미 있게 증가하였으나 tumor necrosis factor ${\alpha}$ ($TNF{\alpha}$)에 의해서는 증가하지 않았다. Prostacyclin $I_2$ ($PGI_2$)는 혈관평활근세포의 증식은 촉진시켰으나 이동은 오히려 억제하였다. prostaglandin $D_2$ ($PGD_2$) 및 prostaglandin $E_2$ ($PGE_2$)는 혈관평활근세포의 증식을 촉진시켰으나 이동에는 영향을 미치지 않았다. $PGF_{2{\alpha}}$는 용량 의존적으로 혈관평활근세포의 증식 및 이동을 촉진시켰고 EC50는 약 $0.1{\mu}M$로 관찰되었다. 혈관평활근세포에서 phospholipase $C-{\beta}3$ ($PLC-{\beta}3$) 아형의 발현은 매우 높았으나 $PLC-{\beta}1$, $PLC-{\beta}2$, 및 $PLC-{\beta}4$의 발현은 관찰되지 않았다. U73122 처리를 통해 PLC의 활성을 억제하면 $PGF_{2{\alpha}}$에 의한 혈관평활근세포의 이동이 억제되었다. 또한 $PLC-{\beta}3$의 발현을 억제하면 $PGF_{2{\alpha}}$에 의한 혈관평활근세포의 증식 및 이동이 억제되었다. 이러한 결과들을 바탕으로 $PGF_{2{\alpha}}$ 는 혈관평활근세포의 증식 및 이동에 중요한 역할을 수행하고, 여기에는 $PLC-{\beta}3$가 필수적인 역할을 담당하고 있음을 제안한다.