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Chinese Medical Journal 2002年第4期115卷 494-497页

作者：尹航汪丽蕙霍勇彭旭夏春芳唐朝枢北京大学第一医院心脏科北京100034

OBJECTIVE: To study the effects of focal adhesion kinase (FAK) phosphorylation on smooth muscle cells (SMCs) adhesion and migration stimulated by fibronectin. METHODS: Adhesion and migration of cultured SMCs were stimulated by different concentrations of fibronectin (FN), FAK and its phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense oligodeoxynucleotides (ODNs) were transfected into SMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation. SMCs adhesion and migration were also measured by morphological enumeration and modified Boyden Chambers, respectively. RESULTS: FAK were expressed when SMCs adhesion and migration were successfully simulated by different concentrations of FN. FAK phosphorylation were detected only at 20 microg/ml FN or more. FAK antisense ODNs were transfected efficiently by cationic lipid and FAK phosphorylation was inhibited substantially. The SMCs migration rate in the 5 - 60 microg/ml FN groups was reduced by 17.89% - 27.67%. Cell migration stimulated by FN at 10, 20, 40 and 60 microg/ml were reduced by 23.26%, 21.63%, 19.31% and 17.88%, respectively (P

关键词： Animals Cell Adhesion Cell Movement Cells, Cultured DNA, Antisense Dose-Response Relationship, Drug Enzyme Activation Fibronectins Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases muscle, smooth, vascular Phosphorylation Protein-Tyrosine Kinase Rats Transfection

The role of membrane potential and calcium kinetic changes in the pathogenesis of vascular hyporeactivity during severe shock

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Chinese Medical Journal 2000年第1期113卷 59-64页

作者：赵克森刘杰金春华 Department of Pathophysiology First Military Medical University Guangzhou 510515 China.

OBJECTIVE: To determine the role of membrane potential and intracellular calcium kinetic changes in producing vascular hyporeactivity during severe hemorrhagic shock. METHODS: Rats were subjected to hemorrhagic shock (HS) for 2 hours. The spinotrapezius muscle was prepared for microscopy and the responses of arterioles in the muscle to norepinephrine (NE) were tested. The resting membrane potentials of isolated arterial strips were measured with a microelectrode. Membrane potential and intracellular Ca2+ ([Ca2+]i) changes in isolated arteriolar smooth muscle cells (ASMCs) were determined with fluorescent probes and a confocal microscopy. RESULTS: The arteriolar resting membrane potential was decreased from -36.7 +/- 6.3 mV in control to -29.2 +/- 5.3 mV concurrent with the increase of vasoreactivity to NE at 20 minutes after HS. At 120 minutes post-HS, the resting potential hyperpolarized to -51.9 +/- 9.1 mV, and NE stimulated [Ca2+]i increase was reduced to 50% of the control values during the appearance of arteriolar hyporeactivity, i.e. the NE threshold of the arteriolar response increased 15 fold 2 hours after the onset of hemorrhage as compared with normal animals. The state of vasoreactivity was closely related to the resting potential of vascular smooth muscle in hemorrhagic shock, with a correlation coefficient of 0.96. Treatment with glybenclamide, a selective blocker of ATP-sensitive K+ (KATP) channels, decreased the resting potential, increased NE-stimulated [Ca2+]i increase, and partially restored vasoreactivity in severe hemorrhagic shock. CONCLUSION: The results suggested that membrane hyperpolarization and the reduction of NE-stimulated [Ca2+]i increase in smooth muscle cells appeared to contribute to the vascular hyporeactivity in hemorrhagic shock. The mechanism is likely to involve in KATP channels.

关键词： Animals Calcium Glyburide Membrane Potentials muscle, smooth, vascular Norepinephrine Potassium Channels Rats Rats, Sprague-Dawley Shock, Hemorrhagic

Effect of arsenic trioxide on inhibition of restenosis after rabbit vascular injury and its mechanism

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Chinese Medical Journal 2002年第11期115卷 1608-1614,144-145页

作者：赵智深黄从新王晶江洪李建军王晞武汉大学人民医院心内科武汉430060

OBJECTIVE: To investigate the effect and mechanism of arsenic trioxide (As(2)O(3)) on the prevention of restenosis after vascular injury. METHODS: Apoptosis induction of As(2)O(3) on cultured rabbit vascular smooth muscle cells (VSMCs) in vitro was observed. Thirty-two New Zealand white rabbits were randomly divided into 2- and 4-wk study groups, and their controls. 10% As(2)O(3) at 2.5 mg x Kg(-1) x d(-1) or 0.9% sodium chloride was intraperitoneally infused for 3 days before left common carotid arteries were denudated with a balloon. After denudation 2- and 4-wk animals were sacrificed for morphometry and immunohistochemical studies on carotid arteries, and for histopathology on liver and kidney. RESULTS: It was shown via cellular morphology and DNA fragments in electrophoresis that promotion of As(2)O(3) on cultured vascular smooth muscle cell apoptosis was dependent upon its concentration and duration. Compared with the control animals, the mean vascular intimal proliferation areas were reduced in 2-wk study animals (P 0.05), while the mean vascular luminal areas were all enlarged in both study groups (all P

关键词： Animals Apoptosis Arsenicals DNA Female Flow Cytometry Male muscle, smooth, vascular Oxides Proto-Oncogene Proteins Proto-Oncogene Proteins c-bcl-2 Rabbits vascular Diseases bcl-2-Associated X Protein

Interleukin-1β,Tumor Necrosis Factor-α and Lipopolysaccharide Induce Expression of Monocyte Chemoattractant Protein-1 in Calf Aortic smooth muscle Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2000年第1期20卷 36-38页

作者：孟峰邓仲端倪娟 Department of Pathology School of Basic Medical Sciences Tongji Medical University Wuhan

Summary: To investigate whether interleukin-1β(IL-1β), tumor necrosis factor-α (TNF-α) and lipopolysaccharide (LPS) induce expression of monocyte chemoattractant protein-1 (MCP-1 ) mRNA and protein in calf aortic smooth muscle cells(SMCs), calf aortic SMCs were cultured by a substrate-attached explant method. The cultured SMCs were used between the third to the fifth passage. After the cells became confluent, the SMCs were exposed to 2 ng/ml IL-1β, 20ng/ml TNF-1α and 100 ng/ml LPS respectively, and the total RNA of SMCs which were incubated for 4 h at 37℃ were extracted from the cells by using guanidinium isothiocyanate method. The expres- ion of MCP-1 mRNA in SMCs was detected by using dot blotting analysis using a probe of γ-32 P- end-labelled 35-mer oligonucleotide. After a 24-h incubation, the media conditioned by the cul- tured SMCs were collected. The MCP-1 protein content in the conditioned media was determined by using sandwich ELISA. The results were as follows: Dot blotting analysis showed that the cul- tured SMCs could express MCP-1 mRNA. After a 4-h exposure to IL-1β, TNF-α and LPS, the MCP-1 mRNA expression in SMCs was increased (3.6-fold, 2. 3-fold and 1. 6-fold, respectively). ELISA showed that the levels of MCP-1 protein in the conditioned media were also increased (2.9- fold, 1.7-fold and 1.1-fold, respectively). The results suggest that calf aortic SMCs could ex- press MCP-1 mRNA and protein. IL-1β and TNF-α can induce strong expression of MCP-1 mRNA and protein, and the former is more effective than the latter.

关键词： interleukin-1β tumor necrosis factor α lipopolysaccaride monocyte chemoattractant protein 1 muscle, smooth, vascular

Mechanism of action of prostaglandin E_2 in the dog skeletal muscle circulation

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Chinese Medical Journal 1998年第10期 81-86页

作者： H. J. Ballard and F. I. Achike

Objective To determine whether prostaglandin E2 (PGE2) influences the dog skeletal muscle circulation by a direct action on the vascular smooth muscle or via pre- or post-synaptic modulation of sympathetic ***　In 18 anaesthetised dogs, a gracilis muscle was vascularly isolated and perfused at constant flow. Sympathetic vasoconstrictor tone on the muscles was reflexly controlled by alterations to the pressure at which the isolated carotid sinuses were perfused. The effects of PGE2 injection into the muscle were compared at low carotid sinus pressure, high carotid sinus pressure, and following denervation of the muscle, with or without noradrenaline ***　At all levels of sympathetic tone, PGE2 produced significantly more vasodilation than the saline vehicle. However, at a carotid sinus pressure of 46.0±2.3 mmHg (1 mmHg=0.133 kPa), PGE2 caused a decrease in femoral arterial perfusion pressure of 52.6±7.1 mmHg, which was significantly greater than the response at a carotid sinus pressure of 208.5±3.7 (33.6±4.2 mmHg decrease) or following denervation (25.6±3.7 mmHg decrease). In a separate group of denervated muscles, PGE2 caused a similar decrease in perfusion pressure in the presence or absence of a noradrenaline ***　PGE2 appears to cause vasodilation through two separate mechanisms: one mechanism involves presynaptic inhibition of sympathetic vasoconstrictor tone, whilst the other is independent of the sympathetic nervous system, and is therefore presumably a direct action on the vascular smooth muscle or endothelium. Under our experimental conditions, both mechanisms contributed equally to the vasodilation.

关键词： Animals Carotid Sinus Dinoprostone Dogs Female Male muscle, Skeletal muscle, smooth, vascular Regional Blood Flow Research Support, Non-U.S. Gov't Vasodilator Agents

Evidence of apoptotic smooth muscle cells in proliferative intima of injured arteries

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Chinese Medical Journal 2000年第1期113卷 10-13页

作者：黄峻尹航冷静姚玉宇姚蓉芬彭韬李晶阁 Department of Cardiology First Affiliated Hospital of Nanjing Medical University Nanjing 210029 China.

OBJECTIVE: To investigate the occurrence and extent of apoptosis in the course of restenosis. METHODS: The experimental models of vessel narrowness and intima thickness were established in minipigs' iliac arteries by balloon injury and specimens were retrieved on the 1st, 3rd, 6th, 12th and 30th days for dynamic observation. Apoptotic smooth muscle cells (SMCs) were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). RESULTS: Apoptotic SMCs occurred only in the thickened intima 12 days after injury accompanied with the proliferative SMCs, the percentage of apoptosis was 1.94% +/- 0.42% on the 12th day and 1.36% +/- 0.31% on the 30th day respectively. The low frequency of apoptosis compared with the proliferative SMCs was a feature in the restenotic pathology. CONCLUSIONS: Apoptosis participates in the pathogenetic process of intimal thickening and its level was low compared with proliferation. The findings suggest that attempts to modulate apoptosis after vessel injury constitute a theoretical approach to the prevention of restenosis.

关键词： Apoptosis Animals Arterial Occlusive Diseases Cell Division Iliac Artery muscle, smooth, vascular Research Support, Non-U.S. Gov't Swine Swine, Miniature

Endothelin-1, an important mitogen of smooth muscle cells of spontaneously hypertensive rats

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Chinese Medical Journal 2002年第5期115卷 750-752,156页

作者：刘革修王华欧大明黄红林廖端芳南华大学药物药理研究所衡阳421001

OBJECTIVE: To study the features of vascular smooth muscle cell (VSMC) proliferation induced by endothelin-1 (ET-1). METHODS: VSMCs of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats were cultured and treated with ET-1. Basic fibroblast growth factor (bFGF) gene expression was measured using both Northern blot and an enzyme-linked immunoassay. RESULTS: ET-1 resulted in an increase in bFGF transcripts at 8 - 24 h; bFGF levels were significantly higher in VSMCs treated with ET-1 than in those not treated. However, VSMCs growth responses in SHR and WKY were different. smooth muscle cells of SHR were hyper-responsive to ET-1. Maximal bFGF mRNA levels were elevated 3.5-fold at 4 h of stimulation in WKY and 8-fold at 8h in SHR4. Moreover, the proliferation of VSMCs induced by ET-1 was inhibited by antisense phosphorothioate oligodeoxynucleotides (10 micromol/L AS-bFGF) but not sense bFGF oligomers at the same concentrations, being reduced by 80% in SHR and 40% in WKY vs control, respectively. Furthermore, the effect of AS-bFGF oligomers on SHR SMC proliferation is significantly greater than on WKY SMC proliferation. CONCLUSION: ET-1 may be required for exaggerated vascular growth responses in SHR and bFGF may be involved.

关键词： Animals Cell Division Cells, Cultured Comparative Study DNA, Antisense Dose-Response Relationship, Drug Endothelin-1 Fibroblast Growth Factor 2 Gene Expression Regulation muscle, smooth, vascular Oligonucleotides RNA, Messenger Rats Rats, Inbred SHR Rats, Inbred WKY Research Support, Non-U.S. Gov't Time Factors