국소마취제가 원발성 고혈압 흰쥐의 흉부대동맥 평활근에 미치는 영향

강성희,장태호,김병권,박진웅,김세환,백운이,홍정길 대한마취과학회 1992 Korean Journal of Anesthesiology Vol.25 No.2

The vascular actions of local anesthetics are important in determining the uptake and distribution of these agents from their site of injection as well as influencing their hemodynamic effects once absorbed. Because of the importance of the endothelium in determining of modulating the vascular response of a wide variety of agents, cumulative dose-dependent vasular effects of lidocaine, mepivacaine and bupivacaine on isolated rings of thoracic aorta in normotensive rats(NTR) and spontaneously hypertensive rats(SHR) were studied in the presence and absence of intact endothelium. The results were as follows ; The body weight of NTR and SHR averaged 274.71±55.80(N = 38) and 241.43±17.73gm(N = 18) and mean arterial pressure was 74.4l±3.60 and 129.34±2.89mmHg respectively. The mean absolute value of the contraction induced by 5×l0^(-6) M phenylephrine was 3.27±0.98(N = 18) and 2.3l±50.64gm(N = 18) with intact endothelium and 3.12±0.92 and 2.46±0.87 gm without intact endothelium in aortic rings of NTR and SHively. In the response to local anesthetics in preparation with resting tension(1.0 gm), lidocaine and mepivacaine in concentration of 10^(-3) to 1.25×10^(-2) M not produced dose dependent contraction in aortic ring with intact endothelium from NTR. but bupivacaine produced dose-dependent contraction in aortic rings with intact endothelium from NTR. In the aortic rings from NTR and SHR previously contracted with phenylephrine, lidocaine in contraction of 10^(-3) to 1.25×10^(-2) M caused dose related relaxation in aortic rings with or without endothelium but in concentration of 10^(-3) to 510^(-3) M, aortic rings with endothelium were more relaxed than those af without endothelium in NTR. In SHR, aortic rings without endothelium in concentration of 5×10^(-3) to 1.25×10 M were more significantly relaxed than those of with endothelium. In aortic rings from NTR previously contracted with phenylephrine, mepivacaine caused dose-related relaxation, which was more profound in SHR. In aortic rings with endothelium froR previously contracted with phenylephrine, bupivacaine in concentration of 10^(-3) to 1.5×10^(-3) M caused a relaxation and in concentration of 2.5×10^(-3) to 7.5×10^(-3) M and 1.25×10^(-3) M caused a relaxation again. But in the aortic rings without intact endothelium, bupivacaine caused dose-related relaxation. In the aortic rings without intact endothelium, bupivacaine caused dose-related relaxation in NTR. In the aortic rings from SHR previously contracted with phenylephrine, bupivacaine caused dose-related relaxation, which was more profound than those of NTR. The local anesthetics appear to exert their relaxant effect on endothelium independently and more profoundly in SHR.

Artemisinin attenuates platelet-derived growth factor BB-induced migration of vascular smooth muscle cells

Kang Pa Lee,Eun-Seok Park,Dae-Eun Kim,In-Sik Park,Jin Tack Kim,Heeok Hong 한국영양학회 2014 Nutrition Research and Practice Vol.8 No.5

BACKGROUND/OBJECTIVES: Artemisinin (AT), an active compound in Arternisia annua, is well known as an anti-malaria drug. It is also known to have several effects including anti-oxidant, anti-inflammation, and anti-cancer activities. To date, the effect of AT on vascular disorders has not been studied. In this study, we investigated the effects of AT on the migration and proliferation of vascular smooth muscle cells (VSMC) stimulated by platelet-derived growth factor BB (PDGF-BB). MATERIALS/METHODS: Aortic smooth muscle cells were isolated from Sprague-Dawley rats. PDGF-BB stimulated VSMC migration was measured by the scratch wound healing assay and the Boyden chamber assay. Cell viability was determined by using an EZ-Cytox Cell Viability Assay Kit. The production of reactive oxygen species (ROS) in PDGF-BB stimulated VSMC was measured through H2DCF-DA staining. We also determined the expression levels of signal proteins relevant to ROS, including measures of extracellular signal-regulated kinase (ERK) 1/2 measured by western blot analysis and matrix metalloproteinase (MMP) 9 measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: AT (10 μM and 30 μM) significantly reduced the proliferation and migration of PDGF-BB stimulated VSMC in a dose-dependent manner. The production of ROS, normally induced by PDGF-BB, is reduced by treatment with AT at both concentrations. PDGF-BB stimulated VSMC treated with AT (10 μM and 30 μM) have reduced phosphorylation of ERK1/2 and inhibited MMP9 expression compared to untreated PDGF-BB stimulated VSMC. CONCLUSIONS: We suggest, based on these results, that AT may exert an anti-atherosclerotic effect on PDGF-BB stimulated VSMCs by inhibiting their proliferation and migration through down-regulation of ERK1/2 and MMP9 phosphorylation.

HSP90 inhibitor, AUY922, debilitates intrinsic and acquired lapatinib-resistant HER2-positive gastric cancer cells

( Kang-seo Park ),( Yong Sang Hong ),( Junyoung Choi ),( Shinkyo Yoon ),( Jihoon Kang ),( Deokhoon Kim ),( Kang-pa Lee ),( Hyeon-su Im ),( Chang Hoon Lee ),( Seyoung Seo ),( Sang-we Kim ),( Dae Ho Lee 생화학분자생물학회(구 한국생화학분자생물학회) 2018 BMB Reports Vol.51 No.12

Human epidermal growth factor receptor 2 (HER2) inhibitors, such as trastuzumab and lapatinib are used to treat HER2-positive breast and gastric cancers. However, as with other targeted therapies, intrinsic or acquired resistance to HER2 inhibitors presents unresolved therapeutic problems for HER2-positive gastric cancer. The present study describes investigations with AUY922, a heat shock protein 90 (HSP90) inhibitor, in primary lapatinib-resistant (ESO26 and OE33) and lapatinib-sensitive gastric cancer cells (OE19, N87, and SNU-216) harboring HER2 amplification/over-expression. In order to investigate whether AUY922 could overcome intrinsic and acquired resistance to HER2 inhibitors in HER2-positive gastric cancer, we generated lapatinib-resistant gastric cancer cell lines (OE19/LR and N87/LR) by continuous exposure to lapatinib in vitro. We found that activation of HER2 and protein kinase B (AKT) were key factors in inducing intrinsic and acquired lapatinib-resistant gastric cancer cell lines, and that AUY922 effectively suppressed activation of both HER2 and AKT in acquired lapatinib-resistant gastric cancer cell lines. In conclusion, AUY922 showed a synergistic anti-cancer effect with lapatinib and sensitized gastric cancer cells with intrinsic resistance to lapatinib. Dual inhibition of the HSP90 and HER2 signaling pathways could represent a potent therapeutic strategy to treat HER2-positive gastric cancer with intrinsic and acquired resistance to lapatinib. [BMB Reports 2018; 51(12): 660-665]

DJ-1 is involved in epigenetic control of sphingosine-1-phosphate receptor expression in vascular neointima formation

Lee, Kang Pa,Baek, Suji,Jung, Seung Hyo,Cui, Long,Lee, Donghyen,Lee, Dong-Youb,Choi, Wahn Soo,Chung, Hyun Woo,Lee, Byeong Han,Kim, Bokyung,Won, Kyung Jong Springer-Verlag 2018 Pfl ugers Arch Vol.470 No.7

Anti-neuroinflammatory effects of ethanolic extract of black chokeberry (Aronia melanocapa L.) in lipopolysaccharide-stimulated BV2 cells and ICR mice

Kang Pa Lee,Nan Hee Choi,Hyun-Soo Kim,Sanghyun Ahn,In-Sik Park,Dea Won Lee 한국영양학회 2018 Nutrition Research and Practice Vol.12 No.1

BACKGROUND/OBJECTIVES: One of the mechanisms considered to be prevalent in the development of Alzheimer’s disease (AD) is hyper-stimulation of microglia. Black chokeberry (Aronia melanocapa L.) is widely used to treat diabetes and atherosclerosis, and is known to exert anti-oxidant and anti-inflammatory effects; however, its neuroprotective effects have not been elucidated thus far. MATERIALS/METHODS: We undertook to assess the anti-inflammatory effect of the ethanolic extract of black chokeberry friut (BCE) in BV2 cells, and evaluate its neuroprotective effect in the lipopolysaccharide (LPS)-induced mouse model of AD. RESULTS: Following stimulation of BV2 cells by LPS, exposure to BCE significantly reduced the generation of nitric oxide as well as mRNA levels of numerous inflammatory factors such as inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), interleukin 1 beta (IL-1β), and tumor necrosis factor alpha (TNF-α). In addition, AD was induced in a mouse model by intraperitoneal injection of LPS (250 μg/kg), subsequent to which we investigated the neuroprotective effects of BCE (50 mg/kg) on brain damage. We observed that BCE significantly reduced tissue damage in the hippocampus by downregulating iNOS, COX-2, and TNF-α levels. We further identified the quinic acids in BCE using liquid chromatography-mass spectrometry (LCMS). Furthermore, we confirmed the neuroprotective effect of BCE and quinic acid on amyloid beta-induced cell death in rat hippocampal primary neurons. CONCLUSIONS: Our findings suggest that black chokeberry has protective effects against the development of AD.

Cytoprotective effect of rhamnetin on miconazole-induced H9c2 cell damage

Kang Pa Lee,Jai-Eun Kim,Won-Hwan Park 한국영양학회 2015 Nutrition Research and Practice Vol.9 No.6

BACKGROUND/OBJECTIVES: Reactive oxygen species (ROS) formation is closely related to miconazole-induced heart dysfunction. Although rhamnetin has antioxidant effects, it remained unknown whether it can protect against miconazole-induced cardiomyocyte apoptosis. Thus, we investigated the effects of rhamnetin on miconazole-stimulated H9c2 cell apoptosis. MATERIALS/METHODS: Cell morphology was observed by inverted microscope and cell viability was determined using a WelCount™ cell proliferation assay kit. Miconazole-induced ROS production was evaluated by fluorescence-activated cell sorting with 6-carboxy-2 ʹ,7ʹ-dichlorofluoroscein diacetate (H₂DCF-DA) stain. Immunoblot analysis was used to determine apurinic/apyrimidinic endonuclease 1 (APE/Ref-1) and cleaved cysteine-aspartic protease (caspase) 3 expression. NADPH oxidase levels were measured using real-time polymerase chain reaction. RESULTS: Miconazole (3 and 10 μM) induced abnormal morphological changes and cell death in H9c2 cells. Rhamnetin enhanced the viability of miconazole (3 μM)-treated cells in a dose-dependent manner. Rhamnetin (1 and 3 μM) treatment downregulated cleaved caspase 3 and upregulated APE/Ref-1 expression in miconazole-stimulated cells. Additionally, rhamnetin significantly reduced ROS generation. CONCLUSIONS: Our data suggest that rhamnetin may have cytoprotective effects in miconazole-stimulated H9c2 cardiomyocytes via ROS inhibition. This effect most likely occurs through the upregulation of APE/Ref-1 and attenuation of hydrogen peroxide levels.

The effect of yacon (Samallanthus sonchifolius) ethanol extract on cell proliferation and migration of C6 glioma cells stimulated with fetal bovine serum

Kang Pa Lee,Nan Hee Choi,Jin Teak Kim,In-Sik Park 한국영양학회 2015 Nutrition Research and Practice Vol.9 No.3

BACKGROUND/OBJECTIVES: Yacon (Samallanthus sonchifolius), a common edible plant grown throughout the world, is well known for its antidiabetic properties. It is also known to have several other pharmacological properties including anti-inflammatory, anti-oxidant, anti-allergic, and anti-cancer effects. To date, the effect of yacon on gliomas has not been studied. In this study, we investigated the effects of yacon on the migration and proliferation of C6 glioma cells stimulated by fetal bovine serum (FBS). MATERIALS/METHODS: Cell growth and proliferation were determined by evaluating cell viability using an EZ-Cytox Cell Viability Assay Kit. FBS-induced migration of C6 glioma cells was evaluated by performing the scratch wound healing assay and the Boyden chamber assay. We also used western blot analysis to determine the expression levels of extracellular signal-regulated kinase 1/2 (ERK1/2), a major regulator of migration and proliferation of glioma cells. Matrix metallopeptidase (MMP) 9 and TIMP-1 levels were measured by performing reverse transcription PCR. RESULTS: Yacon (300 μg/mL) reduced both the FBS-induced proliferation of C6 glioma cells and the dose-dependent migration of the FBS-stimulated C6 cells. FBS-stimulated C6 glioma cells treated with yacon (200 and 300 μg/mL) showed reduced phosphorylation of ERK1/2 and inhibition of MMP 9 expression compared to those shown by the untreated FBS-stimulated C6 cells. In contrast, yacon (200 and 300 μg/mL) induced TIMP-1 expression. CONCLUSIONS: On the basis of these results, we suggest that yacon may exert an anti-cancer effect on FBS-stimulated C6 glioma cells by inhibiting their proliferation and migration. The most likely mechanism for this is down-regulation of ERK1/2 and MMP9 and up-regulation of TIMP-1 expression levels.

The anti-inflammatory effect of Indonesian Areca catechu leaf extract in vitro and in vivo

Kang Pa Lee,Giftania Wardani Sudjarwo,Ji-Su Kim,Septrianto Dirgantara,Won Jai Maeng,Heeok Hong 한국영양학회 2014 Nutrition Research and Practice Vol.8 No.3

BACKGROUND/OBJECTIVES: Overproduction of nitric oxide (NO) by the inducible nitric oxide synthase (iNOS) enzyme can cause inflammation. Cyclooxygenase-2 (COX-2) is also involved in the inflammatory response through regulation of nuclear factor-kappa B (NF-κB). Areca catechu is one of the known fruit plants of the Palmaceae family. It has been used for a long time as a source of herbal medicine in Indonesia. In this study, we explored the effect of Indonesian Areca catechu leaf ethanol extract (ACE) in lipopolysaccharide (LPS)-induced inflammation and carrageenan-induced paw edema models. Recently, this natural extract has been in the spotlight because of its efficacy and limited or no toxic side effects. However, the mechanism underlying its anti-inflammatory effect remains to be elucidated. MATERIALS/METHODS: We measured NO production by using the Griess reagent, and determined the expression levels of inflammation-related proteins, such as iNOS, COX2, and NF-κB, by western blot. To confirm the effect of ACE in vivo, we used the carrageenan-induced paw edema model. RESULTS: Compared to untreated cells, LPS-stimulated RAW 264.7 cells treated with ACE showed reduced NO generation and reduced iNOS and COX-2 expression. We found that the acute inflammatory response was significantly reduced by ACE in the carrageenan-induced paw edema model. CONCLUSION: Taken together, these results suggest that ACE can inhibit inflammation and modulate NO generation via downregulation of iNOS levels and NF-κB signaling in vitro and in vivo. ACE may have a potential medical benefit as an anti-inflammation agent.

Angiotensin II facilitates neointimal formation by increasing vascular smooth muscle cell migration: Involvement of APE/Ref-1-mediated overexpression of sphingosine-1-phosphate receptor 1

Lee, Dong-Youb,Won, Kyung-Jong,Lee, Kang Pa,Jung, Seung Hyo,Baek, Suji,Chung, Hyun Woo,Choi, Wahn Soo,Lee, Hwan Myung,Lee, Byeong Han,Jeon, Byeong Hwa,Kim, Bokyung Elsevier 2018 Toxicology and applied pharmacology Vol.347 No.-

<P><B>Abstract</B></P> <P>Angiotensin II (Ang II) is implicated in the development of cardiovascular disorders including hypertension and atherosclerosis. However, the role of Ang II in the interaction between apurinic/apyrimidinic endonuclease/redox factor-1 (APE/Ref-1) and sphingosine-1-phosphate (S1P) signals in relation to vascular disorders remains to be clarified. This study aimed to determine whether APE/Ref-1 plays a role in epigenetic regulation of the S1P receptor (S1PR) in response to Ang II in vascular smooth muscle cell (VSMC) migration and vascular neointima formation. Ang II augmented the expression of S1PR1 in aortic smooth muscle cells of Sprague Dawley rats (RASMCs), which was attenuated by Ang II receptor (AT) 1 inhibitors, antioxidants, and APE/Ref-1 knockdown with small interference RNA. Ang II stimulation produced H<SUB>2</SUB>O<SUB>2</SUB>, and exogenous H<SUB>2</SUB>O<SUB>2</SUB> elevated S1PR1 expression in RASMCs. Moreover, Ang II caused translocation of cytoplasmic APE/Ref-1 into the nucleus in RASMCs. H3 histone acetylation and APE/Ref-1 binding at the S1PR1 promoter were increased in RASMCs treated with Ang II. In addition, Ang II induced migration in RASMCs, which was suppressed by AT1 and S1PR1 inhibitors. The expression of S1PR1, and colocalization of APE/Ref-1 and acetylated histone H3 in vascular neointima, were greater in Ang II-infused rats compared with a control group. These findings demonstrate that Ang II stimulates the epigenetic regulation of S1PR1 expression via H<SUB>2</SUB>O<SUB>2</SUB>-mediated APE/Ref-1 translocation, which may consequently be involved in Ang II-induced VSMC migration and vascular neointima formation. Therefore, APE/Ref-1-mediated overexpression of S1PR1 may be implicated in the vascular dysfunction evoked by Ang II.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Ang II increased S1PR1 expression and H<SUB>2</SUB>O<SUB>2</SUB> generation in VSMCs. </LI> <LI> H<SUB>2</SUB>O<SUB>2</SUB> elevated S1PR1 expression in VSMCs. </LI> <LI> Ang II epigenetically enhanced S1PR1 expression via APE/Ref-1 translocation by H<SUB>2</SUB>O<SUB>2</SUB>. </LI> <LI> These events may be linked to Ang II-increased VSMC migration and vascular neointima. </LI> </UL> </P>

Epigenetic regulation of <i>Kcna3</i>-encoding Kv1.3 potassium channel by cereblon contributes to regulation of CD4⁺ T-cell activation

Kang, Jung-Ah,Park, Sang-Heon,Jeong, Sang Phil,Han, Min-Hee,Lee, Cho-Rong,Lee, Kwang Min,Kim, Namhee,Song, Mi-Ryoung,Choi, Murim,Ye, Michael,Jung, Guhung,Lee, Won-Woo,Eom, Soo Hyun,Park, Chul-Seung,Pa National Academy of Sciences 2016 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF Vol.113 No.31

<P>The role of cereblon (CRBN) in T cells is not well understood. We generated mice with a deletion in Crbn and found cereblon to be an important antagonist of T-cell activation. In mice lacking CRBN, CD4(+) T cells show increased activation and IL-2 production on T-cell receptor stimulation, ultimately resulting in increased potassium flux and calcium-mediated signaling. CRBN restricts T-cell activation via epigenetic modification of Kcna3, which encodes the Kv1.3 potassium channel required for robust calcium influx in T cells. CRBN binds directly to conserved DNA elements adjacent to Kcna3 via a previously uncharacterized DNA-binding motif. Consequently, in the absence of CRBN, the expression of Kv1.3 is derepressed, resulting in increased Kv1.3 expression, potassium flux, and CD4(+) T-cell hyperactivation. In addition, experimental autoimmune encephalomyelitis in T-cell-specific Crbn-deficient mice was exacerbated by increased T-cell activation via Kv1.3. Thus, CRBN limits CD4(+) T-cell activation via epigenetic regulation of Kv1.3 expression.</P>