骨钙素及Ⅰ型胶原在体外血管钙化细胞中的表达

Expression of osteocalcin and collagen Ⅰ in vitro model of vascular calcification

  • 摘要: 目的:检测骨钙素(osteocalcin OC)及Ⅰ型胶原(collagenⅠ ColⅠ)在体外血管钙化细胞中的表达。方法:建立体外血管钙化细胞模型(钙化BASMCs),比色法检测细胞层钙含量、放免法检测培养上清OC浓度,逆转录聚合酶链反应(RT-PCR)检测OC、ColⅠmRNA表达。结果:钙化BASMCs中钙沉积增加呈时间及剂量依赖性;培养上清OC含量在钙化过程中随时间延长呈升高趋势,至第8天升至最高,继而轻度下降;OC及ColⅠmRNA在钙化BASMCs中表达分别增加128.05%及116.84%。结论:钙化BASMCs中OC及ColⅠ表达增加,是一较好的研究血管钙化的体外模型。

     

    Abstract: Objective:To detect the expression of osteocalcin(OC)and collagenⅠ(ColⅠ)in vitro model of vascular Calcification. Methods:1.Establish a model of vascular calcification in vitro:BASMCs were obtained by a modification of the explant method. Cells of passage 2-8 were cultured in standard medium for 4 days and then switched to calcification medium(containing 10mmo/l Lβ-glycerophosphate [β-GP],10-7 mo/l L insulin and 50μg/L Vit C)for 10 days.The medium was replaced with fresh medium every 2 days;2.Calcium deposition in the cellular layer、OC in cultured supernatant and the expression of OC、ColⅠmRNA in calcified BASMCs.was analyzed. Results:β-GP induced extensive calcium deposition of BASMCs in a dose-dependent and time-dependent manner accompanying increased phosphorus content and decreased calcium content in cultured supernatant;OC in cultured supernatant was increased in a timedependent manner in calcified BASMCs,the highest amount appeared in 8th day and decreased slightly in 10th day;the expression of OC and Col Ⅰ mRNA in calcified BASMCs was increased by 128.05% and 116.84% repectively compared with those of control group.Conclusions: The expression of OC and Col Ⅰ was increased in calcified BASMCs,which shows have a high concordance with cardiovascular ectopic calcification.

     

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