• 동의생리병리학회지
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• 제20권1호
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• pp.163-170
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• 2006

Nardostachyos Rhizoma (N. Rhizoma) belonging to the family Valerianaceae has been anti-arrhythmic effect, and sedation to the central nerve and a smooth muscle. We reported that the water extract of N. Rhizoma induced apoptotic cell death and differentiation in human promyelocytic leukemia (HL-60) cells. Cytotoxicity of N. Rhizoma was detected only in HL-60 cells (IC50 is about 200 ${\mu}g/ml$). The cytotoxic activity of N. Rhizoma in HL-60 cells was increased in a dose-dependent manner. We used several measures of apoptosis to determine whether these processes were involved in N. Rhizoma-induced apoptotic cell death. The high-dose (200 ${\mu}g/ml$) treatment of N. Rhizoma to HL-60 cells showed cell shrinkage, cell membrane blobbing, apoptotic bodies, and the fragmentation of DNA, suggesting that these cells underwent apoptosis. Treatment of HL-60 cells with N. Rhizoma time-dependently induced activation of caspase-3, caspase-8, and caspase-9 and proteolytic cleavage of poly(ADP-ribose) polymerase. Also, we investigated the effect of N. Rhizoma on cellular differentiation and proliferation in HL-60 cells. Differentiation and proliferation of HL-60 cells was determined through expression of CD11b and CD14 surface antigens using flow cytometry and nitroblue tetrazolium (NBT) assay, and through analysis of cell cycle using propidium iodide assay, respectively. N. Rhizoma induced the differentiation of HL-60 at the low-dose (100 ${\mu}g/ml$) treatment, as shown by increased expression of differentiation surface antigen CD11b, but not CDl4 and increased reducing activity of NBT. When HL-60 cells were treated with N. Rhizoma at concentration of $50{\mu}g/ml\;and\;100{\mu}g/ml$, NBT-reducing activities induced approximately 1.5-fold and 20.0-fold as compared with the control. In contrast, HL-60 cells treated with the N. Rhizoma-ATRA combination showed markedly elevated levels of 26.3-fold at $50{\mu}g/ml$ N. Rhizoma-0.1 ${\mu}M$ ATRA combination and 27.5-fold at 50 ${\mu}g/ml$ N. Rhizoma-0.2 ${\mu}M$ ATRA combination than when treated with N. Rhizoma alone or ATRA alone. It may be that N. Rhizoma plays important roles in synergy with ATRA during differentiation of HL-60 cells. DNA flow-cytometry indicated that N. Rhizoma markedly induced a G1 phase arrest of HL-60 cells. N. Rhizoma-treated HL-60 cells increased the cell population in G1 phase from 32.71% to 42.26%, whereas cell population in G2/M and S phases decreased from 23.61% to 10.33% and from 37.78% to 33.98%, respectively. We examined the change in the $p21^{WAF1/Cip1}\;and\;p27^{Kip1}$ proteins, which are the CKIs related with the G1 phase arrest. The expression of the CDK inhibitor $p27^{Kip1},\;but\;not\;p21^{WAF1/Cip1}$ were markedly increased by N. Rhizoma. Taken together, these results demonstrated that N. Rhizoma induces apoptotic cell death through activation of caspase-3, and potently inhibits the proliferation of HL-60 cells via the G1 phase cell cycle arrest in association with $p27^{Kip1}$ and granulocytic differentiation induction .

• 동의생리병리학회지
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• 제21권1호
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• pp.136-144
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• 2007

Biotae Orientalis has been widely used for treatment of relaxion of smooth muscle, gastrointestinal hemorrhage and alopecia in Oriental Medicine. This experiment examined the effect of an extracts, obtained from the acetone and MeOH extracts of dried or fresh Biotae Orientalis, on hair growing activity of the C57BL/6N mice after topical application to skin. First, We examined on hair growth activity of extracts of Biotae Orientalis compare to control and 1% minoxidil groups. Second, We investigated on the number of hair follicle and mast cells after topical application of extracts of the Biotae Orientalis to skin for 16 day. Third, We investigated immunoreactive density of vascular endothelial growth factor(VEGF), protein kinase C-${\alpha}$(PKC-${\alpha}$) and stem(mast) cell factor(SCF) in skin of C57BL/6N mice by immunohistochemical methods. I fourth investigated changes of subpopulation of splenocytes and thymocytes in C57BL/6N mice for 16day using laser flow cytometry. The results were as follows : Hair growing effect of acetone and MeOH extracts of dried and fresh Biotae Orientalis was observed in 70%, 90% and 60% in hair removed skin area in 16 day respectively. Immunoreactive density of VEGF and PKC-${\alpha}$ in skin of experimental groups was weakly stained compare to control group in 10 day. Immunoreactive density of stem cell factor in skin of experimental group was heavily stained compare to control group in 10 day. Splenic TH/TC Iymphocytes of lived MeOH extracts group significantly increased compare to control group. TH cells in thymic T lymphocytes were increased compare to control group. These experiment suggest that acetone and MeOH extracts of Biotae Orientalis may be used for topical treatment of alopecia areata.

• 동의생리병리학회지
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• 제27권4호
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• pp.400-408
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• 2013

This study aimed to investigate the relaxation effects and its underlying mechanisms of Rubus coreanus(RC) extract in contracted rabbit corpus cavernous tissues by phenylephrine(PE) $1{\mu}M$. In order to define the relaxation effects of RC, rabbit corpus cavernous tissues were prepared in $2{\times}2{\times}6mm$ sized strip. The dose-dependent relaxation responses of RC at 0.01-3.0 $mg/m{\ell}$ in contracted strips induced by PE were measured and also observed after endothelial denudation. To analyze the underlying mechanisms of RC-induced relaxation, indomethacin(IM), tetraethylammonium chloride(TEA), $N{\omega}$-nitro-L-arginine (L-NNA), methylene blue(MB) were treated before RC extract infused into precontracted strips induced by PE. To study the effect of RC extract on influx of extracellular $Ca^{2+}$ in corpus cavernous strips, calcium chloride(Ca) 1 mM infused into precontracted strips induced by PE after pretreatment of RC extract in $Ca^{2+}$-free krebs-ringer solution. To investigate cytotoxic activity and nitric oxide(NO) concentration of RC extract on human umbilical vein endothelial cell(HUVEC), cell viability on HUVEC was measured by MTT assay, and NO concentration was measured by Griess reagent system. The cavernous strips were significantly relaxed by RC extract at 1.0 $mg/m{\ell}$, 3.0 $mg/m{\ell}$ and the relaxation responses to RC were inhibited significantly by endothelial disruption. The pretreatment of IM, TEA didn't affect RC extract-induced endothelium-dependent relaxation, but the pretreatment of L-NNA, MB reduced RC extract-induced endothelium-dependent relaxation. When $Ca^{2+}$ was supplied the cavernous strips which were precontracted by PE in a $Ca^{2+}$-free krebs-ringer solution, contraction of strips was increased, but pretreatment of RC inhibited contractile response to $Ca^{2+}$. When RC extract was applicated on HUVEC, NO concentration was increased. Our findings show that RC extract exerts a relaxing effect on corpus cavernosum in part by suppressing influx of extracellular $Ca^{2+}$ through activating the NO-cGMP system.

• Journal of Chest Surgery
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• 제44권6호
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• pp.406-412
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• 2011

Background: Development of thoracic aortic aneurysms and aortic dissections (TAAD) is attributed to unbearable wall tension superimposed on defective aortic wall integrity and impaired aortic repair mechanisms. Central to this repair mechanisms are well-balanced and adequately functional cellular components of the aortic wall, including endothelial cells, smooth muscle cells (SMCs), inflammatory cells, and adventitial fibroblasts. Adventitial fibroblasts naturally produce aortic extracellular matrix (ECM), and, when aortic wall is injured, they can be transformed into SMCs, which in turn are involved in aortic remodeling. We postulated the hypothesis that adventitial fibroblasts in patients with TAAD may have defects in ECM production and SMC transformation. Materials and Methods: Adventitial fibroblasts were procured from the adventitial layer of fresh aortic tissues of patients with TAAD (Group I) and of multi-organ donors (Group II), and 4-passage cell culture was performed prior to the experiment. To assess ECM production, cells were treated with TNF-${\alpha}$ (50 pM) and the expression of MMP-2/MMP-3 was analyzed using western blot technique. To assess SMC transformation capacity, cells were treated with TGF-${\beta}1$ and expression of SM ${\alpha}$-actin, SM-MHC, Ki-67 and SM calponin was evaluated using western blot technique. Fibroblasts were then treated with TGF-${\beta}1$ (10 pM) for up to 10 days with TGF-${\beta}1$ supplementation every 2 days, and the proportion of transformed SMC in the cell line was measured using immunofluorescence assay for fibroblast surface antigen every 2 days. Results: MMP-3 expression was significantly lower in group I than in group II. TGF-${\beta}1$-stimulated adventitial fibroblasts in group I expressed less SM ${\alpha}$-actin, SM-MHC, and Ki-67 than in group II. SM-calponin expression was not different between the two groups. Presence of fibroblast was observed on immunofluorescence assay after more than 6 days of TGF-${\beta}1$ treatment in group I, while most fibroblasts were transformed to SMC within 4 days in group II. Conclusion: ECM production and SMC transformation are compromised in adventitial fibroblasts from patients with TAAD. This result suggests that functional restoration of adventitial fibroblasts could well be a novel approach for the prevention and treatment of TAAD.

• 대한물리치료과학회지
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• 제13권2호
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• pp.99-119
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• 2006

Endothelin (ET) is a 21 amino acid peptide with multifunctional effects on the vasculature as well as a variety of other cell types such as respiratory, gastrointestinal, urogenital, endocrine, central nervous systems, and others. Endothelin has emerged as a modulator by autocrine and paracrine actions for many cellular activities, including vasoconstriction, cell proliferation, hormone production, neurotransmitter and/or neuromodulator. The endothelin family consists of three closely related peptides, ET-1, ET-2, and ET-3 derived from separate genes, such as chromosome 6, 1, and 20, respectively. ET-1 is the predominant isoform produced in the cardiovascular system and about which most is known. Endothelin receptors are seven-transmembrane GTP-binding protein-coupled receptors, which are classified into endothelin-A (ETA) and endothelin-B (ETB) receptors. Interestingly, recent evidence is accumulating to suggest that ET -1 may contribute to a variety of pain states such as allodynia and hyperalgesia in animals and humans. Therefore, in this review the biological characteristics and contraction-related mechanism of endothelin-1 in mammalian cells will be summarized. Especially, we focus on multifunctional roles for ET-1 in noxious stimulation-induced pain for the study of pain specialized physical therapy.

• 폴리머
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• 제37권2호
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• pp.127-134
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• 2013

혈관내피세포는 혈관 안쪽을 덮고 있는 편평한 세포층으로, 혈관의 기능과 혈관평활근세포의 증식을 조절한다. 폴리락타이드글리콜라이드 공중합체(PLGA)는 물성이 좋고 분해속도를 조절하기 좋은 생분해성 합성고분자이며, 여러 형태로 제조하기 쉽다. 누에에서 얻은 실크 피브로인은 18가지 아미노산으로 구성되어 있고 세포의 부착과 세포 기능 유지에 중요하며 화장품, 의료분야 등 다양한 분야에서 응용되고 있다. 본 연구에서는 용매 증발법을 이용하여 0, 10, 20, 40 및 80 wt%의 실크를 이용하여 실크/PLGA 하이브리드 필름을 만들었으며, MTT, SEM, ELISA, 면역세포화학염색법을 실시하였다. 실크/PLGA 하이브리드 필름에서 실크 함량에 따른 인간 대동맥 내피세포의 부착과 증식을 측정한 결과, 40 wt%의 실크/PLGA 하이브리드 필름에서 세포의 부착과 증식이 가장 높았으며, 이런 결과들은 실크가 세포의 증식에 좋은 영향을 미치고 실크/PLGA 하이브리드 필름의 표면이 인간 대동맥 내피세포의 성장에 알맞은 환경이라는 것을 확인할 수 있었다.

• 동의생리병리학회지
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• 제20권1호
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• pp.174-180
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• 2006

We examine the effect of Geiji-Bokryung-Hwan(GBH) on erectile function in a rat model of hypercholesterolemic erectile dysfunction. GBH, a drug preparation consisting of five herbs of Cinnamomi Ramulus (Geiji), Poria Cocos (Bokryun), Mountan Cortex Radicis (Mokdanpi), Paeoniae Radix (Jakyak), and Persicae Semen (Doin) is a traditional Korean herbal medicine that is widely used in the treatment of atherosclerosis-related disorders. In this study, 3-month-old Sprague-Dawley rats were used. The 6 rats control animals were fed a normal diet and the other 18 rats were fed 1% cholesterol diet for 3 months. After 1 months, GBH was added to the drinking water of the treatment group of 12 rats but not the cholesterol only group of 6 rats. Of the 12 rats 6 received 30 mg/kg per day (group 1) and 6 received 60 mg/kg per day (group 2) of GBH. At 3 months erectile function was evaluated with cavernous nerve electrostimulation in all animals. Penile tissues were collected for electron microscopy, and to perform Western blot for endothelial nitric oxide synthase (eNOS), neuronal nitric oxide synthase (nNOS), basic fibroblast growth factor (bFGF) and caveolin-1. Systemic arterial pressure was not significantly different between the animals that were fed the 1% cholesterol diet and the controls. Conversely erectile function was not impaired in the herbal medicine treated rats. Electron microscopy showed many caveolae with fingerlike processes in the cavernous smooth muscle and endothelial cell membranes in control and treated rats but not in the cholesterol only group of rats. Western blot showed differences among groups in protein expression for eNOS, nNOS, caveolin-1 and bFGF protein expression in penile tissue. Increased eNOS and nNOS protein expressions dy high cholesterol diet were significantly decreased in group 1 and group 2. Interestingly, caveolin-1 and bFGF protein expression was significantly higher in groups 1 and 2 than in the cholesterol only and control groups.

• Nutrition Research and Practice
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• 제13권4호
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• pp.302-309
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• 2019

BACKGOURND/OBJECTIVES: Vascular inflammation is an important feature in the atherosclerotic process. Recent studies report that leaves and branches of Carpinus turczaninowii (C. turczaninowii) have antioxidant capacity and exert anti-inflammatory effects. However, no study has reported the regulatory effect of C. turczaninowii extract on the arterial inflammatory response. This study therefore investigated modulation of the arterial inflammatory response after exposure to C. turczaninowii extract, using human aortic vascular smooth muscle cells (HAoSMCs). MATERIALS/METHODS: Scavenging activity of free radicals, total phenolic content (TPC), cell viability, mRNA expressions, and secreted levels of cytokines were measured in LPS-stimulated (10 ng/mL) HAoSMCs treated with the C. turczaninowii extract. RESULTS: C. turczaninowii extract contains high amounts of TPC ($225.6{\pm}21.0mg$ of gallic acid equivalents/g of the extract), as well as exerts time-and dose-dependent increases in strongly scavenged free radicals (average $14.8{\pm}1.97{\mu}g/mL$ $IC_{50}$ at 40 min). Cell viabilities after exposure to the extracts (1 and $10{\mu}g/mL$) were similar to the viability of non-treated cells. Cytokine mRNA expressions were significantly suppressed by the extracts (1 and $10{\mu}g/mL$) at 6 hours (h) after exposure. Interleukin-6 secretion was dose-dependently suppressed 2 h after incubation with the extract, at $1-10{\mu}g/mL$ in non-stimulated cells, and at 5 and $10{\mu}g/mL$ in LPS-stimulated cells. Similar patterns were also observed at 24 h after incubation with the extract (at $1-10{\mu}g/mL$ in non-stimulated cells, and at $10{\mu}g/mL$ in the LPS-stimulated cells). Soluble intracellular vascular adhesion molecules (sICAM-1) secreted from non-stimulated cells and LPS-stimulated cells were similarly suppressed in a dose-dependent manner after 24 h exposure to the extracts, but not after 2 h. In addition, sICAM-1 concentration after 24 h treatment was positively related to IL-6 levels after 2 h and 24 h exposure (r = 0.418, P = 0.003, and r = 0.524, P < 0.001, respectively). CONCLUSIONS: This study demonstrates that C. turczaninowii modulates the arterial inflammatory response, and indicates the potential to be applied as a therapeutic use for atherosclerosis.

• Clinical and Experimental Pediatrics
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• 제52권8호
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• pp.944-952
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• 2009

목 적 : Epithelial to mesenchymal transition (EMT)은 태생기에 있어 필수 불가결한 발달과정일 뿐 아니라, 신 섬유화에 있어서도 중요한 역할을 하며, nestin은 고전적인 줄기세포 표지자로 신 세뇨관 간질 손상에 있어 새로운 표지자로 밝혀지고 있다. 신생 백서 신장에서 Angiotensin (Ang) II가 EMT에 미치는 영향을 알아보고자, 안지오텐신 전환 효소 억제제를 투여한 신생 백서의 신장에서 EMT 표지자 및 nestin의 발현 양상을 조사하였다. 방 법 : 7일 동안 신생 백서에게 enalapril (30 mg/kg/d) 또는 vehicle을 투여하였으며, ${\alpha}-smooth$ muscle actin (SMA), E-cadherin, vimentin 및 nestin에 대한 면역 조직 화학 염색을 시행하였다. 결 과 : enalapril 투여군에서 대조군에 비해 신 피질 및 수질 모두에서 ${\alpha}-SMA$ 발현이 증가하였으며, 이는 확장된 세뇨관 상피 세포에서 뚜렷하였다(P<0.05). E-cadherin 발현은 enalapril 투여군의 신 피질 및 수질의 세뇨관 상피 세포에서 확연히 감소하였다(P<0.05). vimentin 및 nestin 발현은 신 피질에서는 양군간의 차이가 없었으나, 신 수질에서는 enalapril 투여군에서 세뇨관 간질 세포에서 발현이 의미있게 증가하였다(P<0.05). 결 론 : 신생 백서 신장에서 Ang II 억제는 ${\alpha}-SMA$ 발현을 증가시키고, E-cadherin 발현을 감소시킴으로써 발달하는 신장의 EMT를 증가시켰다. Enalapril 투여는 또한 신 수질에서 vimentin과 nestin의 발현을 증가시켰으며, 이는 신생 백서 신장에서의 Ang II 억제로 인한 EMT 과정 중 신 수질의 변화가 더욱 뚜렷한 것을 시사하며, Ang II 억제가 EMT 표지자들의 발현을 다르게 변화시키는 것으로 사료된다.

• Journal of Periodontal and Implant Science
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• 제37권sup2호
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• pp.447-463
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• 2007

치주인대세포는 시험관적 실험에서 광물화 결절형성을 유도할 수 있으므로 광물화 결절형성에 관여하는 유전자들을 특이하게 발현할 것으로 여겨진다. 이에 본 실험은 cDNA microarray를 이용한 동시 유전자분석을 시행하여 치주인대세포의 분화에 의한 광물화 결절형성시 나타나는 유전자의 특징적 발현 양상을 알아보고자 하였다. 교정치료를 목적으로 경북대학교병원에 내원한 환자의 제일소구치를 발치하여 통상적 방법으로 치주인대세포를 분리, 배양하였고, 3세대의 치주인대세포를 사용하여 실험을 시행하였다. 치주인대세포를 100mm 배양접시에 넣고 배양하여 매 2일 마다 배지를 교환해 주고, 10% FBS만을 투여한 대조군으로, ascorbic acid $(50\;{\mu}g/ml)$, ${\beta}-glycerophosphate$ (10 mM) 및 100 nM dexamethasone을 투여한 군을 실험군으로 하였다. 배양된 치주인대세포에 ascorbic acid, ${\beta}-glycerophosphate$, 그리고 dexamethasone을 투여한 실험군에서 21일째 광물화된 결정을 관찰할 수 있었으나 대조군에서는 관찰할 수 없었다. 3063개의 유전자를 분석한 결과 35개 유전자가 대조군에 비해 2배이상 발현이 증가하였고, 38개 유전자는 2배이상 발현이 감소하였다. 형태학적 검사에서 보여준 바와 같이 광물화 형성과정시 관여하는 JGF-2과 IGFBP2와 같은 유전자가 실험군에서 증가하였으며, 세포골격과 세포외기질 형성에 관여하는 proteogycan 1, fibulin-5, keratin 5, ${\beta}-actin$, ${\alpha}-smooth$ muscle actin, capping protein 등도 발현이 실험군에서 증가하였다. 한편 periostin and S100 calcium-binding protein A4는 대조군에서 오히려 높게 나타나므로 이는 배양된 치주인대세포가 그 자체의 표현형을 유지하고 있음을 보여 주고 있다. 그 외 apoptosis를 유발시키는데 관여하는 Dkk-1와 Nip3는 실험군에서 높게 발현되었고, apoptosis를 억제시키는데 관여하는 Btf와 TAX1BP1는 오히려 낮게 발현됨을 알 수 있으므로 이는 실험군에서 치주인대세포가 골아세포로의 분화되었음을 나타낸다.