摘要
目的 :检测骨钙素 (osteocalcinOC)及Ⅰ型胶原 (collagenⅠColⅠ )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 12 8 0 5 %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。
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 10mmol/Lβ glycerophosphate,10 7 mol/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 time dependent manner in calcified BASMCs,the highest amount appeared in 8th day and decreased slightly in 10th day; the expression of OC and ColⅠmRNA in calcified BASMCs was increased by 128 05% and 116 84% repectively compared with those of control group Conclusions: The expression of OC and ColⅠwas increased in calcified BASMCs, which shows have a high concordance with cardiovascular ectopic calcification
出处
《军医进修学院学报》
CAS
2003年第1期61-63,共3页
Academic Journal of Pla Postgraduate Medical School
基金
国家自然科学基金资助项目 ( 3 9770 869)
