Background In addition to elevated concentrations of cytokines, patients with congestive endothelial dysfunction and increased plasma concentrations of adhesion molecules heart failure （CHF） show ke intercellular ad...Background In addition to elevated concentrations of cytokines, patients with congestive endothelial dysfunction and increased plasma concentrations of adhesion molecules heart failure （CHF） show ke intercellular adhesion molecule-1 （ICAM-1）. Furthermore, the concentration of cardiotrophin-1 （CT-1） - a cytokine of the interleukin-6 superfamily - is increased in CHF. We tested the hypothesis whether CT-1 is able to induce ICAM-1 in human umbilical vein endothelial cells （HUVEC）. Furthermore we examined the signalling mechanisms of CT-1 mediated ICAM-1 expression. Methods Confluent layers of HUVEC were incubated with increasing concentrations of CT-1 （5 to 100 ng/ml） for different periods. ICAM-1 mRNA was determined by real-time polymerase chain reaction （PCR） and ICAM-1 surface expression by fluorescence-activated cell sorter （FACS） analysis and soluble ICAM-1 （slCAM-1） in the culture supernatant by enzyme linked immunosorbent assay （ELISA）. To clarify the signalling pathway of CT-1 induced ICAM-1 expression we used various inhibitors of possible signal transducing molecules, electromobility shift assay （EMSA） and Western blot analysis. Results CT-1 induced ICAM-1 mRNA （1.8±0.8 fold increase compared to unstimulated cells after 6 hours） and protein （1.4±0.2 fold increase compared to unstimulated cells after 48 hours） in HUVEC in a time- and concentration-dependent manner. EMSA experiments show that CT-1 causes nuclear factor （NF） κB activation. Because parthenolide could inhibit CT-1 induced ICAM-1 expression NFκB activation is required in this pathway. CT-1 did not activate extracellular signal regulated kinases （ERK）, c-Jun N-terminal kinase （JNK） and p38. Conclusion CT-1 is able to induce ICAM-1 in endothelial cells by NFκB activation. These results may explain in part elevated ICAM-1 concentrations in patients with CHF and endothelial dysfunction.展开更多
The aim of this paper was to investigate the effects of resistin on human umbilical vein endothelial cells(HUVECs),and to explore its role and mechanism of action in atherosclerosis.HUVECs were incubated with recombin...The aim of this paper was to investigate the effects of resistin on human umbilical vein endothelial cells(HUVECs),and to explore its role and mechanism of action in atherosclerosis.HUVECs were incubated with recombinant human resistin(0,50,100 ng/mL)for 24 h.ICAM-1,VCAM-1 and reactive oxygen species(ROS)were assayed by flow cytometer.ET-1,eNOS and iNOS mRNA expression were measured by semi-quantitative RT-PCR.Incubation of HUVECs with resistin resulted in an increase in ICAM-1 expression and ET-1 mRNA expression.However,resistin had no effect on VCAM-1 expression and ROS release.eNOS and iNOS mRNA expression were not altered by resistin stimulation.Adipokine resistin exerted a direct effect in promoting HUVEC dysfunction by promoting ICAM-1 and ET-1 expression.These data suggest that adipocyteendothelium cross-talk might play an important role in the pathogenesis of cardiovascular disease in diabetes mellitus.展开更多
目的探讨Destrin蛋白在氧化型低密度脂蛋白(oxygenized low density lipoprotein,oxLDL)损伤人颈动脉内皮细胞过程中的作用和机制。方法人颈动脉内皮细胞给予剂量梯度的oxLDL(0、5、25、100μg/ml)刺激24h,检测细胞中Destrin蛋白的表达...目的探讨Destrin蛋白在氧化型低密度脂蛋白(oxygenized low density lipoprotein,oxLDL)损伤人颈动脉内皮细胞过程中的作用和机制。方法人颈动脉内皮细胞给予剂量梯度的oxLDL(0、5、25、100μg/ml)刺激24h,检测细胞中Destrin蛋白的表达变化;通过RNA干扰的方式下调颈动脉内皮细胞中Destrin的表达,观察其对oxLDL刺激诱导的颈动脉内皮细胞损伤的影响,同时检测单核细胞趋化蛋白1(MCP-1)和细胞间黏附分子1(ICAM-1)的表达水平。结果与oxLDL刺激对照RNAi比较,oxLDL刺激RNAi MCP-1和ICAM-1蛋白表达明显增加[(734.2±113.4)pg/ml vs(502.1±96.7)pg/ml,(152.5±22.0)pg/ml vs(94.2±16.5)pg/ml,P<0.05];细胞活力明显减低[0.41±0.05 vs 0.62±0.06,P<0.05],细胞凋亡明显增高[(50±10)%vs(32±8)%,P<0.05]。结论oxLDL损伤颈动脉内皮细胞的过程中,Destrin蛋白表达下调是oxLDL的损伤机制之一。展开更多
文摘Background In addition to elevated concentrations of cytokines, patients with congestive endothelial dysfunction and increased plasma concentrations of adhesion molecules heart failure （CHF） show ke intercellular adhesion molecule-1 （ICAM-1）. Furthermore, the concentration of cardiotrophin-1 （CT-1） - a cytokine of the interleukin-6 superfamily - is increased in CHF. We tested the hypothesis whether CT-1 is able to induce ICAM-1 in human umbilical vein endothelial cells （HUVEC）. Furthermore we examined the signalling mechanisms of CT-1 mediated ICAM-1 expression. Methods Confluent layers of HUVEC were incubated with increasing concentrations of CT-1 （5 to 100 ng/ml） for different periods. ICAM-1 mRNA was determined by real-time polymerase chain reaction （PCR） and ICAM-1 surface expression by fluorescence-activated cell sorter （FACS） analysis and soluble ICAM-1 （slCAM-1） in the culture supernatant by enzyme linked immunosorbent assay （ELISA）. To clarify the signalling pathway of CT-1 induced ICAM-1 expression we used various inhibitors of possible signal transducing molecules, electromobility shift assay （EMSA） and Western blot analysis. Results CT-1 induced ICAM-1 mRNA （1.8±0.8 fold increase compared to unstimulated cells after 6 hours） and protein （1.4±0.2 fold increase compared to unstimulated cells after 48 hours） in HUVEC in a time- and concentration-dependent manner. EMSA experiments show that CT-1 causes nuclear factor （NF） κB activation. Because parthenolide could inhibit CT-1 induced ICAM-1 expression NFκB activation is required in this pathway. CT-1 did not activate extracellular signal regulated kinases （ERK）, c-Jun N-terminal kinase （JNK） and p38. Conclusion CT-1 is able to induce ICAM-1 in endothelial cells by NFκB activation. These results may explain in part elevated ICAM-1 concentrations in patients with CHF and endothelial dysfunction.
基金This study was supported by the Foundation of the Ministry of Health of China(No.000099009)Foundation of Ministry of Education of China(No.002000001).
文摘The aim of this paper was to investigate the effects of resistin on human umbilical vein endothelial cells(HUVECs),and to explore its role and mechanism of action in atherosclerosis.HUVECs were incubated with recombinant human resistin(0,50,100 ng/mL)for 24 h.ICAM-1,VCAM-1 and reactive oxygen species(ROS)were assayed by flow cytometer.ET-1,eNOS and iNOS mRNA expression were measured by semi-quantitative RT-PCR.Incubation of HUVECs with resistin resulted in an increase in ICAM-1 expression and ET-1 mRNA expression.However,resistin had no effect on VCAM-1 expression and ROS release.eNOS and iNOS mRNA expression were not altered by resistin stimulation.Adipokine resistin exerted a direct effect in promoting HUVEC dysfunction by promoting ICAM-1 and ET-1 expression.These data suggest that adipocyteendothelium cross-talk might play an important role in the pathogenesis of cardiovascular disease in diabetes mellitus.
文摘目的探讨Destrin蛋白在氧化型低密度脂蛋白(oxygenized low density lipoprotein,oxLDL)损伤人颈动脉内皮细胞过程中的作用和机制。方法人颈动脉内皮细胞给予剂量梯度的oxLDL(0、5、25、100μg/ml)刺激24h,检测细胞中Destrin蛋白的表达变化;通过RNA干扰的方式下调颈动脉内皮细胞中Destrin的表达,观察其对oxLDL刺激诱导的颈动脉内皮细胞损伤的影响,同时检测单核细胞趋化蛋白1(MCP-1)和细胞间黏附分子1(ICAM-1)的表达水平。结果与oxLDL刺激对照RNAi比较,oxLDL刺激RNAi MCP-1和ICAM-1蛋白表达明显增加[(734.2±113.4)pg/ml vs(502.1±96.7)pg/ml,(152.5±22.0)pg/ml vs(94.2±16.5)pg/ml,P<0.05];细胞活力明显减低[0.41±0.05 vs 0.62±0.06,P<0.05],细胞凋亡明显增高[(50±10)%vs(32±8)%,P<0.05]。结论oxLDL损伤颈动脉内皮细胞的过程中,Destrin蛋白表达下调是oxLDL的损伤机制之一。
