Summary： Although previous reports showed dmg-eluting stent （DES） could effectively inhibit neointima formation, in-stent restenosis （ISR） remains an important obstacle. The purpose of this study was to investiga...Summary： Although previous reports showed dmg-eluting stent （DES） could effectively inhibit neointima formation, in-stent restenosis （ISR） remains an important obstacle. The purpose of this study was to investigate different effects of paclitaxel on proliferation and cell cycle regulators between vascular smooth muscle cells （VSMCs） and vascular endothelial cells （VECs） of rats in vitro. The cultured VSMCs and VECs of rats from the same tissues were examined by using immunohistochemistry, flow cytometry and Western blotting in control and paclitaxel-treated groups. The results showed paclitaxel could effectively inhibit proliferation of VSMCs and VECs. However, as compared with VECs, prolif- eration of VSMCs in paclitaxel-treated group decreased less rapidly. The percentage of cells in G0-G1 and G2-M phases was reduced, and that in S phase increased after treatment for 72 h. The expression of cyclin D1 and B1, p27 and PCNA in VSMCs of paclitaxel-treated group was up-regulated, but that of p21 down-regulated as compared with VECs. It is concluded that there are significant differences in the expression of cell cycle regulators and proliferation rate between paclitaxel-treated VSMCs and paclitaxel-treated VECs, suggesting that the G1 S checkpoint regulated by paclitaxel may play a critical role in the development of complications of DES, which provides new strategies for treatments of ISR.展开更多
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 stim...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展开更多
Background Batroxobin (BX),a serine protease used in defibrinogenation and thrombolysis,also has an effect on c-fos gene and growth factor. This study attempted to determine the effects of BX on the proliferation of v...Background Batroxobin (BX),a serine protease used in defibrinogenation and thrombolysis,also has an effect on c-fos gene and growth factor. This study attempted to determine the effects of BX on the proliferation of vascular smooth muscle cells (VSMCs) and calcium metabolism. Methods VSMCs were treated with BX at concentrations of 0.1,0.3,or 1.0 mmol/L and cell numbers were determined at 0,24,48,and 72 hours. Intracellular calcium concentration ([Ca 2+ ]_i) was measured using direct fluorescence methods. Results BX was found to suppress proliferation of VSMCs in a dose-dependent fashion with inhibition rates of 18% and 31% by 48 and 72 hours,respectively. In addition,BX decreases basal [Ca 2+ ]_i significantly. The basal level in untreated cells was 162.7±33.8 nmol/L,and decreased to 131.5±27.7 nmol/L,128.3±28.5 nmol/L,and 125.6±34.3 nmol/L with the three concentrations of BX,respectively. Noradrenaline (NE)-induced [Ca 2+ ]_i stimulation was also attenuated by BX (0.1 mmol/L BX,20%±8% inhibition; 0.3 mmol/L BX,54%±11% inhibition; 1.0 mmol/L BX,62%±15% inhibition). The ability of NE to stimulate [Ca 2+ ]_i was attenuated in cultures in Ca 2+ -free medium,as was the ability of BX to blunt NE-induced stimulation. Conclusion These findings demonstrate that BX can effectively inhibit proliferation of VSMCs,probably by blocking the release and uptake of Ca 2+ ,thus influencing [Ca 2+ ]_i.展开更多
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 trea...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.展开更多
To explore the possibility of hydrogen sulfide (H 2S) as a messenger molecule in cardiovascular system, the authors discovered that H 2S (5×10 -5 -5×10 -4 mol·L -1 )exerted an effect on inhibiting endot...To explore the possibility of hydrogen sulfide (H 2S) as a messenger molecule in cardiovascular system, the authors discovered that H 2S (5×10 -5 -5×10 -4 mol·L -1 )exerted an effect on inhibiting endothelin 1 induced proliferation of cultured vascular smooth muscle cells (VSMCs) of rats in vitro . The 3H TdR incorporation decreased by 16.8%～37.4% in H 2S treated VSMCs as compared with the controls ( P <0.01). The inhibitory effect was found to be associated with reduced activity of MAPK. The authors also observed that endogenous H 2S levels markedly increased in vessels of rats with either endotoxic shock or septic shock [H 2S level (pmol·min -1 ·mg -1 ):tail artery (16.18±2.06) vs (8.12±0.55);mesenteric artery (10.17±1.11) vs (6.19 ±0.55);pulmonary artery(11.38±1.24) vs (5.27±0.51); aorta(6.21±0.48) vs (4.10± 0.28), P < 0.01 ]. The above findings suggested that H 2S might play an important role in the regulation of cardiovascular pathophysiologic events.展开更多
目的:研究盐酸吗啡对肠道平滑肌运动的影响及其作用机制。方法:制备家兔离体小肠,记录小肠平滑肌发展张力、静止张力、收缩频率,观察盐酸吗啡对肠肌运动的影响。并进一步测定不同浓度的盐酸吗啡灌胃前后小鼠小肠推进运动的变化。结果:5 ...目的:研究盐酸吗啡对肠道平滑肌运动的影响及其作用机制。方法:制备家兔离体小肠,记录小肠平滑肌发展张力、静止张力、收缩频率,观察盐酸吗啡对肠肌运动的影响。并进一步测定不同浓度的盐酸吗啡灌胃前后小鼠小肠推进运动的变化。结果:5 m g/L、10 m g/L、30 m g/L的盐酸吗啡作用于离体肠肌,对家兔小肠发展张力均有明显的抑制作用(P<0.05)。纳洛酮可削弱盐酸吗啡对肠肌发展张力的抑制作用,吗啡对肠肌发展张力的抑制作用从78.7%±13.4%至87.8%±11.2%(P<0.05)。阿托品可阻断盐酸吗啡对肠肌发展张力的抑制作用,吗啡对肠肌发展张力的抑制百分比从77.2%±12.1%至103.7%±12.8%(P<0.05)。而酚妥拉明则增强盐酸吗啡对肠肌发展张力的抑制作用,吗啡对肠肌发展张力的抑制百分比从79.2%±11.8%至69.8%±15.3%(P<0.05)。用不同浓度(75、150、300 m g/L)的盐酸吗啡灌胃后小鼠小肠推进均减慢,推进率分别为54.9%±15.5%、47.7%±14.3%、37.1%±5.8%,与生理盐水灌胃组比较有显著性意义(P<0.05)。结论:盐酸吗啡在离体与在体水平对肠肌运动均有抑制作用,其机制可能与阿片受体、胆碱能受体、肾上腺素能受体有关。展开更多
基金supported by grants from National Natural Science Foundation of China(No.81030021)National Basic Research Program of China(No.2011CB504403)
文摘Summary： Although previous reports showed dmg-eluting stent （DES） could effectively inhibit neointima formation, in-stent restenosis （ISR） remains an important obstacle. The purpose of this study was to investigate different effects of paclitaxel on proliferation and cell cycle regulators between vascular smooth muscle cells （VSMCs） and vascular endothelial cells （VECs） of rats in vitro. The cultured VSMCs and VECs of rats from the same tissues were examined by using immunohistochemistry, flow cytometry and Western blotting in control and paclitaxel-treated groups. The results showed paclitaxel could effectively inhibit proliferation of VSMCs and VECs. However, as compared with VECs, prolif- eration of VSMCs in paclitaxel-treated group decreased less rapidly. The percentage of cells in G0-G1 and G2-M phases was reduced, and that in S phase increased after treatment for 72 h. The expression of cyclin D1 and B1, p27 and PCNA in VSMCs of paclitaxel-treated group was up-regulated, but that of p21 down-regulated as compared with VECs. It is concluded that there are significant differences in the expression of cell cycle regulators and proliferation rate between paclitaxel-treated VSMCs and paclitaxel-treated VECs, suggesting that the G1 S checkpoint regulated by paclitaxel may play a critical role in the development of complications of DES, which provides new strategies for treatments of ISR.
文摘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
基金ThisstudywassupportedbytheNationalNaturalScienceFoundationofChina (No 3 0 0 0 0 163 )
文摘Background Batroxobin (BX),a serine protease used in defibrinogenation and thrombolysis,also has an effect on c-fos gene and growth factor. This study attempted to determine the effects of BX on the proliferation of vascular smooth muscle cells (VSMCs) and calcium metabolism. Methods VSMCs were treated with BX at concentrations of 0.1,0.3,or 1.0 mmol/L and cell numbers were determined at 0,24,48,and 72 hours. Intracellular calcium concentration ([Ca 2+ ]_i) was measured using direct fluorescence methods. Results BX was found to suppress proliferation of VSMCs in a dose-dependent fashion with inhibition rates of 18% and 31% by 48 and 72 hours,respectively. In addition,BX decreases basal [Ca 2+ ]_i significantly. The basal level in untreated cells was 162.7±33.8 nmol/L,and decreased to 131.5±27.7 nmol/L,128.3±28.5 nmol/L,and 125.6±34.3 nmol/L with the three concentrations of BX,respectively. Noradrenaline (NE)-induced [Ca 2+ ]_i stimulation was also attenuated by BX (0.1 mmol/L BX,20%±8% inhibition; 0.3 mmol/L BX,54%±11% inhibition; 1.0 mmol/L BX,62%±15% inhibition). The ability of NE to stimulate [Ca 2+ ]_i was attenuated in cultures in Ca 2+ -free medium,as was the ability of BX to blunt NE-induced stimulation. Conclusion These findings demonstrate that BX can effectively inhibit proliferation of VSMCs,probably by blocking the release and uptake of Ca 2+ ,thus influencing [Ca 2+ ]_i.
文摘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.
文摘To explore the possibility of hydrogen sulfide (H 2S) as a messenger molecule in cardiovascular system, the authors discovered that H 2S (5×10 -5 -5×10 -4 mol·L -1 )exerted an effect on inhibiting endothelin 1 induced proliferation of cultured vascular smooth muscle cells (VSMCs) of rats in vitro . The 3H TdR incorporation decreased by 16.8%～37.4% in H 2S treated VSMCs as compared with the controls ( P <0.01). The inhibitory effect was found to be associated with reduced activity of MAPK. The authors also observed that endogenous H 2S levels markedly increased in vessels of rats with either endotoxic shock or septic shock [H 2S level (pmol·min -1 ·mg -1 ):tail artery (16.18±2.06) vs (8.12±0.55);mesenteric artery (10.17±1.11) vs (6.19 ±0.55);pulmonary artery(11.38±1.24) vs (5.27±0.51); aorta(6.21±0.48) vs (4.10± 0.28), P < 0.01 ]. The above findings suggested that H 2S might play an important role in the regulation of cardiovascular pathophysiologic events.
文摘目的:研究盐酸吗啡对肠道平滑肌运动的影响及其作用机制。方法:制备家兔离体小肠,记录小肠平滑肌发展张力、静止张力、收缩频率,观察盐酸吗啡对肠肌运动的影响。并进一步测定不同浓度的盐酸吗啡灌胃前后小鼠小肠推进运动的变化。结果:5 m g/L、10 m g/L、30 m g/L的盐酸吗啡作用于离体肠肌,对家兔小肠发展张力均有明显的抑制作用(P<0.05)。纳洛酮可削弱盐酸吗啡对肠肌发展张力的抑制作用,吗啡对肠肌发展张力的抑制作用从78.7%±13.4%至87.8%±11.2%(P<0.05)。阿托品可阻断盐酸吗啡对肠肌发展张力的抑制作用,吗啡对肠肌发展张力的抑制百分比从77.2%±12.1%至103.7%±12.8%(P<0.05)。而酚妥拉明则增强盐酸吗啡对肠肌发展张力的抑制作用,吗啡对肠肌发展张力的抑制百分比从79.2%±11.8%至69.8%±15.3%(P<0.05)。用不同浓度(75、150、300 m g/L)的盐酸吗啡灌胃后小鼠小肠推进均减慢,推进率分别为54.9%±15.5%、47.7%±14.3%、37.1%±5.8%,与生理盐水灌胃组比较有显著性意义(P<0.05)。结论:盐酸吗啡在离体与在体水平对肠肌运动均有抑制作用,其机制可能与阿片受体、胆碱能受体、肾上腺素能受体有关。
