Aortic valve calcification is a common disease in the elderly, but its cellular and molecular mechanisms are not clear. In order to verify the hypothesis that Wnt/β-catenin signaling pathway is involved in the proces...Aortic valve calcification is a common disease in the elderly, but its cellular and molecular mechanisms are not clear. In order to verify the hypothesis that Wnt/β-catenin signaling pathway is involved in the process of calcification of aortic valve, porcine aortic valve interstitial cells（VICs） were isolated, cultured and stimulated with oxidized low density lipoprotein（ox-LDL） for 48 h to induce the differentiation of VICs into osteoblast-like cells. The key proteins and genes of Wnt/β-catenin signaling pathway, such as glycogen synthase kinase 3β（GSK-3β） and β-catenin, were detected by using Western blotting and real-time polymerase chain reaction（PCR）. The results showed that the VICs managed to differentiate into osteoblast-like cells after the stimulation with ox-LDL and the levels of proteins and genes of GSK-3β and β-catenin were increased significantly in VICs after stimulation for 48 h（P0.05）. It is suggested that Wnt/β-catenin signaling pathway may play a key role in the differentiation of VICs into osteoblast-like cells and make great contribution to aortic valve calcification.展开更多
目的研究白细胞介素17（IL-17）是否促进主动脉瓣膜钙化形成及其可能机制。方法利用体外主动脉瓣膜间质细胞培养技术,瓣膜间质细胞培养传代3-5次后,采用简单随机抽样法分为两组：（1）对照组,细胞培养基中添加2 ml RPMI-1640完全培养液;...目的研究白细胞介素17（IL-17）是否促进主动脉瓣膜钙化形成及其可能机制。方法利用体外主动脉瓣膜间质细胞培养技术,瓣膜间质细胞培养传代3-5次后,采用简单随机抽样法分为两组：（1）对照组,细胞培养基中添加2 ml RPMI-1640完全培养液;（2）实验组,细胞培养基中添加2 ml RPMI-1640完全培养液和IL-17（50 ng/ml）。继续孵育48 h后,用细胞茜素红钙染色检测两组瓣膜间质细胞钙化情况,用Western blot和RT-PCR检测两组瓣膜间质细胞碱性磷酸酶（ALP）及骨形态蛋白2（BMP-2）的表达情况。结果实验组细胞中明显钙结节形成,对照组细胞中少量钙结节形成;与对照组比较,实验组细胞ALP和BMP-2蛋白（0.741±0.063比0.184±0.032;0.900±0.060比0.396±0.030,均为P〈0.01）与基因（0.236±0.004比0.106±0.003;0.523±0.052比0.194±0.047,均为P〈0.01）表达量均明显升高。结论 IL-17可促进主动脉瓣膜间质细胞向成骨样细胞转化。展开更多
基金supported by the National Natural Science Foundation of China(No.81070190)
文摘Aortic valve calcification is a common disease in the elderly, but its cellular and molecular mechanisms are not clear. In order to verify the hypothesis that Wnt/β-catenin signaling pathway is involved in the process of calcification of aortic valve, porcine aortic valve interstitial cells（VICs） were isolated, cultured and stimulated with oxidized low density lipoprotein（ox-LDL） for 48 h to induce the differentiation of VICs into osteoblast-like cells. The key proteins and genes of Wnt/β-catenin signaling pathway, such as glycogen synthase kinase 3β（GSK-3β） and β-catenin, were detected by using Western blotting and real-time polymerase chain reaction（PCR）. The results showed that the VICs managed to differentiate into osteoblast-like cells after the stimulation with ox-LDL and the levels of proteins and genes of GSK-3β and β-catenin were increased significantly in VICs after stimulation for 48 h（P0.05）. It is suggested that Wnt/β-catenin signaling pathway may play a key role in the differentiation of VICs into osteoblast-like cells and make great contribution to aortic valve calcification.
文摘目的研究白细胞介素17（IL-17）是否促进主动脉瓣膜钙化形成及其可能机制。方法利用体外主动脉瓣膜间质细胞培养技术,瓣膜间质细胞培养传代3-5次后,采用简单随机抽样法分为两组：（1）对照组,细胞培养基中添加2 ml RPMI-1640完全培养液;（2）实验组,细胞培养基中添加2 ml RPMI-1640完全培养液和IL-17（50 ng/ml）。继续孵育48 h后,用细胞茜素红钙染色检测两组瓣膜间质细胞钙化情况,用Western blot和RT-PCR检测两组瓣膜间质细胞碱性磷酸酶（ALP）及骨形态蛋白2（BMP-2）的表达情况。结果实验组细胞中明显钙结节形成,对照组细胞中少量钙结节形成;与对照组比较,实验组细胞ALP和BMP-2蛋白（0.741±0.063比0.184±0.032;0.900±0.060比0.396±0.030,均为P〈0.01）与基因（0.236±0.004比0.106±0.003;0.523±0.052比0.194±0.047,均为P〈0.01）表达量均明显升高。结论 IL-17可促进主动脉瓣膜间质细胞向成骨样细胞转化。
