Objective： To test the influence of homocysteine on the production and activation of matrix metalloproteinase-2 （MMP-2） and tissue inhibitors of matrix metalloproteinase-2 （TIMP-2） and on cell migration of cultur...Objective： To test the influence of homocysteine on the production and activation of matrix metalloproteinase-2 （MMP-2） and tissue inhibitors of matrix metalloproteinase-2 （TIMP-2） and on cell migration of cultured rat vascular smooth muscle cells （VSMCs）. Also, to explore whether rosuvastatin can alter the abnormal secretion and activation of MMP-2 and TIMP-2 and migration of VSMCs induced by homocysteine. Methods： Rat VSMCs were incubated with different concentrations of homocysteine （50-5000 μmol/L）. Western blotting and gelatin zymography were used to investigate the expressions and activities of MMP-2 and TIMP-2 in VSMCs in culture medium when induced with homocysteine for 24, 48, and 72 h. Transwell chambers were employed to test the migratory ability of VSMCs when incubated with homocysteine for 48 h. Different concentrations of rosuvastatin （10^-9-10^-5 mol/L） were added when VSMCs were induced with 1 000 pmol/L homocysteine. The expressions and activities of MMP-2 and TIMP-2 were examined after incubating for 24, 48, and 72 h, and the migration of VSMCs was also examined after incubating for 48 h. Results： Homocysteine （50-1000 μmol/L） increased the production and activation of MMP-2 and expression of TIMP-2 in a dose-dependent manner. However, when incubated with 5000 pmol/L homocysteine, the expression of MMP-2 was up-regulated, but its activity was down-regulated. Increased homocysteine-induced production and ac- tivation of MMP-2 were reduced by rosuvastatin in a dose-dependent manner whereas secretion of TIMP-2 was not significantly altered by rosuvastatin. Homocysteine （50-5000 μmol/L） stimulated the migration of VSMCs in a dose-dependent manner, but this effect was eliminated by rosuvastatin. Conclusions： Homocysteine （50-1000 μmol/L） significantly increased the production and activation of MMP-2, the expression of TIMP-2, and the migration of VSMCs in a dose-dependent manner. Additional extracellular rosuvastatin can decrease the excessive expression an展开更多
目的:(1)观察补肾活血通络法对动脉粥样硬化易损斑块的稳定作用及其稳定斑块的可能机制;(2)探讨血清和斑块局部受激活调节正常T细胞表达和分泌因子(Regulated on Activated Normal T cell Expressed and Secreted factor,RANTES)水平与...目的:(1)观察补肾活血通络法对动脉粥样硬化易损斑块的稳定作用及其稳定斑块的可能机制;(2)探讨血清和斑块局部受激活调节正常T细胞表达和分泌因子(Regulated on Activated Normal T cell Expressed and Secreted factor,RANTES)水平与血管平滑肌细胞(vascular smooth muscle cells,VSMCs)凋亡程度的相关性。方法:将健康雄性新西兰兔34只,随机分成空白对照组(K组)和造模组(B组)。空白对照组8只,给予普通饲料喂养18周。造模组(B组)26只,给予高脂饲料喂养,实验开始第1周一次性注射牛血清白蛋白,实验第10周,随机抽取2只兔做病理检测证实模型组已形成粥样斑块。其余B组兔随机分为模型对照组、辛伐他汀组、健脑软脉方组。辛伐他汀组、健脑软脉方组分别予以辛伐他汀、健脑软脉方水煎剂干预,模型对照组予以生理盐水灌胃,持续8周,实验结束,于处死动物前24 h和48 h 2次给予药物触发易损斑块(中国斑点蝰蛇毒和组胺)。空白对照组(K组)按上述给药时间和部位给予同等剂量无菌生理盐水注射。实验终点处死实验兔,开胸迅速分离主动脉全长,检测各项指标。结果:模型对照组的主动脉病变程度、血脂水平、血清RANTES水平、斑块内RANTES m RAN和蛋白表达水平、VSMCs凋亡率与健脑软脉方组、辛伐他汀组、空白对照组相比有显著差异(P<0.05),健脑软脉方组和辛伐他汀组与空白对照组相比有显著差异(P<0.05),健脑软脉方组和辛伐他汀组之间无显著差异(P>0.05)。结论:本研究表明补肾活血通络法可能通过调节血脂水平,抑制炎症反应,抑制平滑肌细胞凋亡,从而抑制模型兔动脉粥样硬化斑块的不稳定性进展。展开更多
目的观察模拟失重2周后,大鼠脑动脉血管平滑肌细胞(vascular smooth muscle cells,VSMCs)L型电压依赖性钙离子通道(L-type voltage dependent calcium channel,CaL)功能的改变,以及钙通道激动剂BayK8644对通道电流的影响。方法以尾部悬...目的观察模拟失重2周后,大鼠脑动脉血管平滑肌细胞(vascular smooth muscle cells,VSMCs)L型电压依赖性钙离子通道(L-type voltage dependent calcium channel,CaL)功能的改变,以及钙通道激动剂BayK8644对通道电流的影响。方法以尾部悬吊大鼠模型模拟失重的影响。采用全细胞膜片钳记录模式,以Ba2+作为载流子,记录2周模拟失重后大鼠脑动脉VSMCs的CaL电流及钙通道激动剂Bay K 8644对其的影响,并测定相应的稳态激活与失活曲线及有关参数。结果与对照组相比,模拟失重2周后悬吊组大鼠已出现了典型的模拟失重效应,悬吊组大鼠脑动脉VSMCs的CaL的电流密度显著增加(P<0.05),且对钙通道激动剂Bay K 8644更敏感(P<0.05)。此外,与对照组相比,悬吊组大鼠脑动脉VSMCs的膜电容与接入电阻、CaL稳态失活曲线和稳态激活曲线等通道动力学特征无显著性改变。结论模拟失重2周可引起大鼠脑动脉平滑肌细胞CaL通道功能增强,这可能是模拟失重导致大鼠脑动脉血管收缩反应性增强的因素之一。展开更多
基金Project supported by the Health Ministry Scientific Research Fund of China (No. WKJ2011-2-018)the Zhejiang Provincial Natural Science Foundation of China (No. Y2100535)+3 种基金the Key Social Development Project of Zhejiang Province (No. 2010A23010)the Science and Technology Projects of Shaoxing (No. 2011A23011)the Science and Technology Plan Project of Zhejiang Province (No. 2012C33040)the Zhejiang Provincial Program for the Cultivation of High-Level Innovative Health Talents, China
文摘Objective： To test the influence of homocysteine on the production and activation of matrix metalloproteinase-2 （MMP-2） and tissue inhibitors of matrix metalloproteinase-2 （TIMP-2） and on cell migration of cultured rat vascular smooth muscle cells （VSMCs）. Also, to explore whether rosuvastatin can alter the abnormal secretion and activation of MMP-2 and TIMP-2 and migration of VSMCs induced by homocysteine. Methods： Rat VSMCs were incubated with different concentrations of homocysteine （50-5000 μmol/L）. Western blotting and gelatin zymography were used to investigate the expressions and activities of MMP-2 and TIMP-2 in VSMCs in culture medium when induced with homocysteine for 24, 48, and 72 h. Transwell chambers were employed to test the migratory ability of VSMCs when incubated with homocysteine for 48 h. Different concentrations of rosuvastatin （10^-9-10^-5 mol/L） were added when VSMCs were induced with 1 000 pmol/L homocysteine. The expressions and activities of MMP-2 and TIMP-2 were examined after incubating for 24, 48, and 72 h, and the migration of VSMCs was also examined after incubating for 48 h. Results： Homocysteine （50-1000 μmol/L） increased the production and activation of MMP-2 and expression of TIMP-2 in a dose-dependent manner. However, when incubated with 5000 pmol/L homocysteine, the expression of MMP-2 was up-regulated, but its activity was down-regulated. Increased homocysteine-induced production and ac- tivation of MMP-2 were reduced by rosuvastatin in a dose-dependent manner whereas secretion of TIMP-2 was not significantly altered by rosuvastatin. Homocysteine （50-5000 μmol/L） stimulated the migration of VSMCs in a dose-dependent manner, but this effect was eliminated by rosuvastatin. Conclusions： Homocysteine （50-1000 μmol/L） significantly increased the production and activation of MMP-2, the expression of TIMP-2, and the migration of VSMCs in a dose-dependent manner. Additional extracellular rosuvastatin can decrease the excessive expression an
文摘目的:(1)观察补肾活血通络法对动脉粥样硬化易损斑块的稳定作用及其稳定斑块的可能机制;(2)探讨血清和斑块局部受激活调节正常T细胞表达和分泌因子(Regulated on Activated Normal T cell Expressed and Secreted factor,RANTES)水平与血管平滑肌细胞(vascular smooth muscle cells,VSMCs)凋亡程度的相关性。方法:将健康雄性新西兰兔34只,随机分成空白对照组(K组)和造模组(B组)。空白对照组8只,给予普通饲料喂养18周。造模组(B组)26只,给予高脂饲料喂养,实验开始第1周一次性注射牛血清白蛋白,实验第10周,随机抽取2只兔做病理检测证实模型组已形成粥样斑块。其余B组兔随机分为模型对照组、辛伐他汀组、健脑软脉方组。辛伐他汀组、健脑软脉方组分别予以辛伐他汀、健脑软脉方水煎剂干预,模型对照组予以生理盐水灌胃,持续8周,实验结束,于处死动物前24 h和48 h 2次给予药物触发易损斑块(中国斑点蝰蛇毒和组胺)。空白对照组(K组)按上述给药时间和部位给予同等剂量无菌生理盐水注射。实验终点处死实验兔,开胸迅速分离主动脉全长,检测各项指标。结果:模型对照组的主动脉病变程度、血脂水平、血清RANTES水平、斑块内RANTES m RAN和蛋白表达水平、VSMCs凋亡率与健脑软脉方组、辛伐他汀组、空白对照组相比有显著差异(P<0.05),健脑软脉方组和辛伐他汀组与空白对照组相比有显著差异(P<0.05),健脑软脉方组和辛伐他汀组之间无显著差异(P>0.05)。结论:本研究表明补肾活血通络法可能通过调节血脂水平,抑制炎症反应,抑制平滑肌细胞凋亡,从而抑制模型兔动脉粥样硬化斑块的不稳定性进展。
文摘目的观察模拟失重2周后,大鼠脑动脉血管平滑肌细胞(vascular smooth muscle cells,VSMCs)L型电压依赖性钙离子通道(L-type voltage dependent calcium channel,CaL)功能的改变,以及钙通道激动剂BayK8644对通道电流的影响。方法以尾部悬吊大鼠模型模拟失重的影响。采用全细胞膜片钳记录模式,以Ba2+作为载流子,记录2周模拟失重后大鼠脑动脉VSMCs的CaL电流及钙通道激动剂Bay K 8644对其的影响,并测定相应的稳态激活与失活曲线及有关参数。结果与对照组相比,模拟失重2周后悬吊组大鼠已出现了典型的模拟失重效应,悬吊组大鼠脑动脉VSMCs的CaL的电流密度显著增加(P<0.05),且对钙通道激动剂Bay K 8644更敏感(P<0.05)。此外,与对照组相比,悬吊组大鼠脑动脉VSMCs的膜电容与接入电阻、CaL稳态失活曲线和稳态激活曲线等通道动力学特征无显著性改变。结论模拟失重2周可引起大鼠脑动脉平滑肌细胞CaL通道功能增强,这可能是模拟失重导致大鼠脑动脉血管收缩反应性增强的因素之一。
