Hypoxic pulmonary vasoconstriction and vascular contractility in monocrotaline-induced pulmonary arterial hypertensive rats

김혜진,유해영 대한약리학회 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.6

Pulmonary arterial hypertension (PAH) is a progressive disease characterized by vascular remodeling of pulmonary arteries (PAs) and increased vascular resistance in the lung. Monocrotaline (MCT), a toxic alkaloid, is widely used for developing rat models of PAH caused by injury to pulmonary endothelial cells; however, characteristics of vascular functions in MCT-induced PAH vary and are not fully understood. Here, we investigated hypoxic pulmonary vasoconstriction (HPV) responses and effects of various vasoconstrictors with isolated/perfused lungs of MCT-induced PAH (PAH-MCT) rats. Using hematoxylin and eosin staining, we confirmed vascular remodeling (i.e., medial thickening of PA) and right ventricle hypertrophy in PAH-MCT rats. The basal pulmonary arterial pressure (PAP) and PAP increase by a raised flow rate (40 mL/min) were higher in the PAH-MCT than in the control rats. In addition, both high K+ (40 mM KCl)- and angiotensin II-induced PAP increases were higher in the PAH-MCT than in the control rats. Surprisingly, application of a nitric oxide synthase inhibitor, L-NG-Nitroarginine methyl ester (L-NAME), induced a marked PAP increase in the PAH-MCT rats, suggesting that endothelial functions were recovered in the three-week PAH-MCT rats. In addition, the medial thickening of the PA was similar to that in chronic hypoxia-induced PAH (PAH-CH) rats. However, the HPV response (i.e., PAP increased by acute hypoxia) was not affected in the MCT rats, whereas HPV disappeared in the PAH-CH rats. These results showed that vascular contractility and HPV remain robust in the MCT-induced PAH rat model with vascular remodeling.

Hypoxic pulmonary vasoconstriction and vascular contractility in monocrotaline-induced pulmonary arterial hypertensive rats

Hae Jin Kim,Hae Young Yoo 대한생리학회-대한약리학회 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.6

Pulmonary arterial hypertension (PAH) is a progressive disease characterized by vascular remodeling of pulmonary arteries (PAs) and increased vascular resistance in the lung. Monocrotaline (MCT), a toxic alkaloid, is widely used for developing rat models of PAH caused by injury to pulmonary endothelial cells; however, characteristics of vascular functions in MCT-induced PAH vary and are not fully understood. Here, we investigated hypoxic pulmonary vasoconstriction (HPV) responses and effects of various vasoconstrictors with isolated/perfused lungs of MCT-induced PAH (PAH-MCT) rats. Using hematoxylin and eosin staining, we confirmed vascular remodeling (i.e., medial thickening of PA) and right ventricle hypertrophy in PAH-MCT rats. The basal pulmonary arterial pressure (PAP) and PAP increase by a raised flow rate (40 mL/min) were higher in the PAH-MCT than in the control rats. In addition, both high K⁺ (40 mM KCl)- and angiotensin II-induced PAP increases were higher in the PAH-MCT than in the control rats. Surprisingly, application of a nitric oxide synthase inhibitor, L-N^G-Nitroarginine methyl ester (L-NAME), induced a marked PAP increase in the PAH-MCT rats, suggesting that endothelial functions were recovered in the three-week PAH-MCT rats. In addition, the medial thickening of the PA was similar to that in chronic hypoxia-induced PAH (PAH-CH) rats. However, the HPV response (i.e., PAP increased by acute hypoxia) was not affected in the MCT rats, whereas HPV disappeared in the PAH-CH rats. These results showed that vascular contractility and HPV remain robust in the MCT-induced PAH rat model with vascular remodeling.

Hypoxic pulmonary vasoconstriction and vascular contractility in monocrotaline-induced pulmonary arterial hypertensive rats

Kim, Hae Jin,Yoo, Hae Young The Korean Society of Pharmacology 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.6

Pulmonary arterial hypertension (PAH) is a progressive disease characterized by vascular remodeling of pulmonary arteries (PAs) and increased vascular resistance in the lung. Monocrotaline (MCT), a toxic alkaloid, is widely used for developing rat models of PAH caused by injury to pulmonary endothelial cells; however, characteristics of vascular functions in MCT-induced PAH vary and are not fully understood. Here, we investigated hypoxic pulmonary vasoconstriction (HPV) responses and effects of various vasoconstrictors with isolated/perfused lungs of MCT-induced PAH (PAH-MCT) rats. Using hematoxylin and eosin staining, we confirmed vascular remodeling (i.e., medial thickening of PA) and right ventricle hypertrophy in PAH-MCT rats. The basal pulmonary arterial pressure (PAP) and PAP increase by a raised flow rate (40 mL/min) were higher in the PAH-MCT than in the control rats. In addition, both high $K^+$ (40 mM KCl)- and angiotensin II-induced PAP increases were higher in the PAH-MCT than in the control rats. Surprisingly, application of a nitric oxide synthase inhibitor, L-$N^G$-Nitroarginine methyl ester (L-NAME), induced a marked PAP increase in the PAH-MCT rats, suggesting that endothelial functions were recovered in the three-week PAH-MCT rats. In addition, the medial thickening of the PA was similar to that in chronic hypoxia-induced PAH (PAH-CH) rats. However, the HPV response (i.e., PAP increased by acute hypoxia) was not affected in the MCT rats, whereas HPV disappeared in the PAH-CH rats. These results showed that vascular contractility and HPV remain robust in the MCT-induced PAH rat model with vascular remodeling.

The Role of 4-hydroxynonenal for Activation of Vascular Smooth Muscle Cell

이지영 대한시과학회 2019 대한시과학회지 Vol.21 No.2

Purpose : Retinal diseases (RD), including proliferative diabetic retinopathy, are among the most important eye diseases leading to vision loss in industrialized countries. In this review, the importance of 4-hydroxynonal (HNE) in association with vascular smooth muscle cell (VSMC) activation leading to vascular remodeling, especially for oxidative stress- related retinal degeneration, is illustrated. Methods : RD is characterized by abnormal angiogenesis associated with an increase in cell proliferation and apoptosis. Angiogenesis in proliferative diabetic retinopathy is a complex multistep phenomenon consisting of the sprouting and the growth of new capillary blood vessels starting from the preexisting ones. Therefore, we investigated the action of HNE in VSMC activation causing vascular remodeling. The important role of HNE as a mediator of oxidative stress-related activation of VSMCs will be also discussed. Results : HNE increased the nitro oxide generation in VSMC. HNE seems to cause cellular structural perturbations and various oxidative stress-related degenerative processes, including vascular dysfunction. HNE also, increased the protein kinase B (Akt) phosphorylation. Oxidative stress by HNE could be mediated an induction of Akt phosphorylation and cell proliferation. Conclusion : Oxidative stress induced by HNE may play a critical role in the pathogenesis of retinal atherosclerosis and vascular disturbance, as angiogenesis. The apparent ability of HNE for activation of VSMC may significantly contribution the promotion and progression of vascular remodeling in oxidative-stress related retinal degeneration.

인간 기관지 평활근 세포에서 Dexamethasone의 PDGF와 TGF-β에 의해 증강된 Vascular Endothelial Growth Factor 생성 억제 효과

유명환 ( Myung Hwan Yoo ),심정연 ( Jung Yeon Shim ),서보길 ( Bo Gil Seo ),김덕수 ( Deok Soo Kim ),심재원 ( Jae Won Shim ),정혜림 ( Hye Lim Jung ),박문수 ( Moon Soo Park ) 대한소아알레르기호흡기학회(구 대한소아알레르기 및 호흡기학회) 2006 소아알레르기 및 호흡기학회지 Vol.16 No.2

목 적 : 인간 기관지 평활근 세포는 천식에서 기관지 운동의 톤을 조절할 뿐만 아니라 기도의 면역조절과 기도개형에서 중요한 역할을 담당하고 있다. VEGF는 다양한 기능을 가지고 있는 사이토카인으로 천식양 표현형을 유도하며 염증, 부종, 혈관 개형, 점액질 화생(mucus metaplasia), 상피세포하 섬유화, 기도 항원 감작 그리고 항원유도 Th2 염증 반응을 일으킨다. 또한 TGF-β는 기도개형에서 인간 기관지 평활근세포의 성장을 조절하는 중요한 사이토카인으로 알려져 있으나 천식 기도개형에서의 역할은 명확하지 않다. 본 연구에서는 배양된 기관지 평활 근 세포에서 dexamethasone이 PDGF와 TGF-β의 자극에 의한 VEGF의 분비에 어떠한 영향을 미치는지 알아보고자 하였다. 방 법 : 10% FCS-DMEM 배지에서 배양된 사람 기관지 평활근세포를 48시간 동안 무혈청 배지에서 성장을 정지시킨 후 dexamethasone (10(-5), 10(-6) M)와 TGF-β(10 ng/mL)로 전처치하였다. 16시간 후 PDGF(20 ng/mL)로 자극하여 24시간 뒤 배양 상층액을 추출 후 VEGF의 ELISA 측정 전까지 영하 80도에 보관하였다. 결 과 : 1)PDGF 자극 후 VEGF의 분비는 대조군에 비하여 의미있는 상승을 보였다(803.14±155.15 pg/mL vs. 376.53±59.41 pg/mL, P<0.01). 2) TGF-β 전처치 후 PDGF로 자극하였을 경우에 TGF-β로 전처치하지 않았을 경우보다 의미있는 VEGF의 상승을 보였다(1,318.96±184.77 pg/mL vs. 803.14±155.15 pg/mL, P< 0.01). 3) Dexamethasone(10(-5), 10(-6) M)은 PDGF로 자극했을 경우와(208.76±78.56 pg/mL vs. 803.14±155.15 pg/mL, 303.56±39.29 pg/mL vs. 803.14±155.15 pg/mL respectively, P< 0.01), TGF-β로 전처치 후 PDGF로 자극했을 경우 모두에서 VEGF의 생성을 억제했다(751.91±209.61 pg/mL vs. 1,318.96±184.77 pg/mL, 204.5±78.56 pg/mL vs. 1,318.96±184.77 pg/ mL respectively, P<0.01). 결 론 : PDGF와 TGF-β는 VEGF 분비를 증가시킴으로써 기도개형에 관여하는 중요한 사이토카인으로 생각되며 글루코코르티코이드는 기관지 평활근세포에서 VEGF의 생성을 조절함으로써 천식 기도개형에 특징 중의 하나인 신생혈관 형성을 억제할 수 있다고 사료된다. Purpose : Human bronchial smooth muscle cell(HBSMC) plays an important role in the remodeling of the airways in asthma. Vascular endothelial growth factor(VEGF) is a multifunctional cytokine, which induces edema, angiogenesis, vascular remodeling, mucus metaplasia, subepithelial fibrosis, and antigen-induced Th2 inflammation. Transforming growth factor-beta(TGF-β) is a growth modulator of HBSMC and an important cytokine in airway remodeling. We investigated the effect of dexamethasone on the release of VEGF from HBSMC stimulated with platelet-derived growth factor(PDGF) and TGF-β. Methods : HBSMC cultured in 10 percent FCS-DMEM media was growth-arrested in serum-deprived medium for 48 hours. Dexamethasone and TGF-β were added and incubated for 16 hours before stimulation with PDGF. After 24 hours of stimulation, culture medium was harvested and stored at -80℃ until ELISA for VEGF was performed. Results : The release of VEGF was significantly increased after stimulation with PDGF (P<0.01). The production of VEGF pretreated with TGF-β before stimulation with PDGF was higher than those without TGF-β pretreatment(P<0.01). Dexamethasone suppressed the release of VEGF in HBSMC stimulated with PDGF(P<0.01), TGF-β and PDGF(P<0.01). Conclusion : PDGF and TGF-β may be one of the key mediators in inducing airway remodeling and glucocorticoid, and can be used as useful therapies to prevent airway vascular remodeling by modulating the VEGF on airway smooth muscle cells. [Pediatr Allergy Respir Dis(Korea) 2006;16:142-149]

Osteopontin plays a key role in vascular smooth muscle cell proliferation via EGFR-mediated activation of AP-1 and C/EBPβ pathways

Lee, S.J.,Baek, S.E.,Jang, M.A.,Kim, C.D. Academic Press 2016 PHARMACOLOGICAL RESEARCH Vol.108 No.-

Osteopontin (OPN) is known as an active player in the progression of vascular remodeling diseases, however, the precise role in the proliferation of vascular smooth muscle cells (VSMC) is unclear. Thus, this study investigated the role of OPN in VSMC proliferation induced by 4-hydroxynonenal (HNE), and identified the intracellular signaling pathways involved in 4-HNE-induced OPN production. In VSMC primary cultured from rat thoracic aorta as well as in VSMC in the media of aorta, HNE enhanced OPN expression in concentration-dependent manners. Both the proliferation of cultured VSMC and PCNA positive cells in aortic tissues were also increased by HNE, which were attenuated in OPN-deficient cells and aortic tissues isolated from OPN-deficient mice, indicating a pivotal role of OPN in HNE-induced VSMC proliferation. In the promoter assay, HNE increased OPN promoter activity, which was attenuated when the regions harboring AP-1 and C/EBPβ binding sites were mutated. The increased bindings of AP-1 and C/EBPβ to the OPN promoter were also demonstrated by ChIP analysis. In addition, the increases in both OPN expression and the activities of AP-1 and C/EBPβ by HNE were attenuated by AG1478, an EGFR antagonist. Based on these results, it was suggested that HNE induced OPN expression in VSMC via signaling pathways involving AP-1 and C/EBPβ, leading to increases in VSMC proliferation and subsequent vascular remodeling.

HMGB1 increases RAGE expression in vascular smooth muscle cells via ERK and p-38 MAPK-dependent pathways

장은정,김희정,백승은,전은영,김지원,김주연,김치대 대한약리학회 2022 The Korean Journal of Physiology & Pharmacology Vol.26 No.5

The increased expression of receptors for advanced glycation endproduct (RAGE) is known as a key player in the progression of vascular remodeling. However, the precise signal pathways regulating RAGE expression in vascular smooth muscle cells (VSMCs) in the injured vasculatures are unclear. Given the importance of mitogen-activated protein kinase (MAPK) signaling in cell proliferation, we investigated the importance of MAPK signaling in high-mobility group box 1 (HMGB1)- induced RAGE expression in VSMCs. In HMGB1 (100 ng/ml)-stimulated human VSMCs, the expression of RAGE mRNA and protein was increased in association with an increase in AGE-induced VSMC proliferation. The HMGB1-induced RAGE expression was attenuated in cells pretreated with inhibitors for ERK (PD98059, 10 μM) and p38 MAPK (SB203580, 10 μM) as well as in cells deficient in ERK and p38 MAPK using siRNAs, but not in cells deficient of JNK signaling. In cells stimulated with HMGB1, the phosphorylation of ERK, JNK, and p38 MAPK was increased. This increase in ERK and p38 MAPK phosphorylation was inhibited by p38 MAPK and ERK inhibitors, respectively, but not by JNK inhibitor. Moreover, AGE-induced VSMC proliferation in HMGB1-stimulated cells was attenuated in cells treated with ERK and p38 MAPK inhibitors. Taken together, our results indicate that ERK and p38 MAPK signaling are involved in RAGE expression in HMGB1-stimulated VSMCs. Thus, the ERK/p38 MAPKRAGE signaling axis in VSMCs was suggested as a potential therapeutic target for vascular remodeling in the injured vasculatures.

The effects of combined treatment of losartan and ramipril on hypertension and related complications

박현수,이도형,한주희,Jung Sang-Hyuk,Lee Miji,장근우,명창선 한국약제학회 2020 Journal of Pharmaceutical Investigation Vol.50 No.6

Purpose Polypharmacy has been an important strategy for the management of hypertension. The aim of this study was to investigate the effect of combined treatment of losartan with ramipril on hypertension, myocardial infarction and vascular remodeling in disease animal models. Methods In 8-telemetered spontaneous hypertensive rats (SHRs) orally treated with various doses of drugs or vehicle by cross-over protocol, systolic blood pressure (SBP), mean arterial pressure (MAP) and heart rate (HR) were measured. The infarct size was measured in rats subjected to 30 min occlusion of the left anterior descending coronary artery (LAD) followed by 150 min reperfusion in myocardial ischemia/reperfusion (MI/R) rats injected drugs or vehicle once through femoral vein 15 min before inducing ischemia. The neointima formation and DNA synthesis in intima and media layer of the blood vessels were measured in cuff-induced vascular injury mice orally treated with drugs for 2 weeks. Results As compared with single treatment, the combination therapy of both drugs significantly decreased SBP and MAP without significant reflex tachycardia. The combined treatment of both drugs significantly increased the expression of endothelial nitric oxide synthase (eNOS) in coronary arterial tissues including area at occlusion. The combination therapy significantly decreased the thickness of vascular media layer and the number of bromodeoxyuridine (BrdU)-positive cells in both intima and media. Conclusion The present data suggest that the combination therapy with fixed doses of losartan and ramipril may exert greater BP-lowering effects with protection against myocardial infarction and vascular remodeling.

The Effect of Interleukin-4 and Amphiregulin on the Proliferation of Human Airway Smooth Muscle Cells and Cytokine Release

심정연,박상욱,김덕수,심재원,정혜림,박문수 대한의학회 2008 Journal of Korean medical science Vol.23 No.5

Airway smooth muscle (ASM) hyperplasia and angiogenesis are important features associated with airway remodeling. We investigated the effect of IL-4 and amphiregulin, an epidermal growth factor family member, on the proliferation of human ASM cells and on the release of vascular endothelial growth factor (VEGF) and monocyte chemotactic protein (MCP)-1 from human ASM cells. Human ASM cells were growth-arrested for 48 hr and incubated with platelet-derived growth factor (PDGF)- BB, interleukin (IL)-4, amphiregulin, and VEGF to evaluate cell proliferation. The cells were treated with PDGF, IL-4 and amphiregulin to evaluate the release of VEGF, MCP-1. IL-4 suppressed unstimulated and PDGF-stimulated ASM cell proliferation. Amphiregulin stimulated ASM cell proliferation in a dose-dependent manner. VEGF did not have any influence on ASM cell proliferation. IL-4 stimulated VEGF secretion by the ASM cells in a dose-dependent manner and showed added stimulatory effects when co-incubated with PDGF. Amphiregulin did not promote VEGF secretion. IL-4 and amphiregulin showed no stimulatory effects on MCP-1 secretion. The results of this study showed that IL-4 had bifunctional effects on airway remodeling, one was the suppression of the proliferation of the ASM cells and the other was the promotion of VEGF release by the ASM cells, and amphiregulin can promote human ASM cell proliferation.

Targeting abnormal airway vascularity as a therapeutical strategy in asthma

PARK, Hee Sun,KIM, Sun Young,KIM, So Ri,LEE, Yong Chul Blackwell Publishing Asia 2010 RESPIROLOGY Vol.15 No.3

<P>ABSTRACT</P><P>Asthma is a chronic inflammatory disease of airways, characterized by airway hyperresponsiveness and airflow limitation with acute bronchoconstriction, swelling of the airway wall, chronic mucus plug formation and airway wall remodelling. Functional and structural changes in the vasculature of asthmatic airways have been documented, and the signalling mechanisms are complex and have recently attracted much attention. The vascular changes may affect inflammatory cell recruitment, airway hyperresponsiveness and the regulation of airway calibre, and further, the level of disease control. Many critical factors are involved in the pathophysiological regulation of vascular changes in bronchial asthma, and the actions of these factors must be very carefully orchestrated. By better understanding the complicated actions of each factor, we may be able to advance further in asthma treatment.</P>