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Cholestane-3beta, 5alpha, 6beta-triol promotes vascular smooth muscle cells calcification.

机译:Cholestane-3beta,5alpha,6beta-triol促进血管平滑肌细胞钙化。

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摘要

Oxysterols found in atherosclerotic plaque may be associated with vascular calcification. We investigated the effect of oxysterol cholestane-3beta, 5alpha, 6beta-triol (Triol) on in vitro calcification of rat vascular smooth muscle cells (VSMCs). In vitro calcification was induced by incubation of VSMCs with beta-glycerophosphate. Calcifying nodule formation, calcium deposition in extracellular matrix, and alkaline phosphatase (ALP) activity were measured as indices of calcification. Because apoptotic bodies can serve as nucleation sites for calcification, apoptosis of calcifying VSMCs was determined by Hoechst 33258 staining, TUNEL, and FITC-labeled annexin V/PI double staining. The calcium deposition and ALP activity in calcifying VSMCs were much higher than those in non-calcifying VSMCs. Triol increased calcifying nodule formation, calcium deposition, ALP activity, and apoptosis of nodular cells in calcifying VSMCs. As determined by 2,7-dichlorofluorescein fluorescence, Triol induced the generation of reactive oxygen species (ROS) in calcifying VSMCs dose- and time-dependently. Triol-induced increases in calcium deposition, ALP activity, apoptosis, and ROS generation were all attenuated by antioxidant vitamin C plus vitamin E (VC + VE). The results demonstrated that Triol promoted VSMCs calcification through direct increase of ALP activity and apoptosis, probably by ROS-related mechanism.
机译:在动脉粥样硬化斑块中发现的氧固醇可能与血管钙化有关。我们调查了氧固醇胆甾醇3beta,5alpha,6beta三醇(Triol)对大鼠血管平滑肌细胞(VSMCs)体外钙化的影响。通过将VSMC与β-甘油磷酸一起孵育来诱导体外钙化。测量钙化结节的形成,钙在细胞外基质中的沉积以及碱性磷酸酶(ALP)的活性,作为钙化的指标。因为凋亡小体可以作为钙化的成核位点,所以通过Hoechst 33258染色,TUNEL和FITC标记的Annexin V / PI双重染色来确定钙化VSMC的凋亡。钙化VSMC中的钙沉积和ALP活性远高于非钙化VSMC中的钙沉积和ALP活性。三醇可增加钙化VSMC中钙化结节的形成,钙沉积,ALP活性和结节细胞的凋亡。由2,7-二氯荧光素荧光确定,Triol可以剂量和时间依赖性地钙化VSMC,从而诱导了活性氧(ROS)的生成。抗氧化剂维生素C加维生素E(VC + VE)减弱了三醇诱导的钙沉积,ALP活性,细胞凋亡和ROS生成的增加。结果表明,Triol通过直接增加ALP活性和凋亡来促进VSMC钙化,可能是由于ROS相关机制。

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