体外培养主动脉细胞钙化作用研究  

STUDIES ON THE MECHANISM OF ARTERIAL IN VITRO CALCIFICATION

作　　者：崔晓暄[1] 王士雯[2] 齐鹏[2] 

机构地区：[1]空军总医院,北京100036 [2]解放军总医院老年心血管病研究所,北京100853

出　　处：《中国应用生理学杂志》2001年第3期271-274,共4页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助课题 (3 9770 86 9)

摘　　要：目的 :初步探讨主动脉钙化的机制。方法 :外殖块法分离培养家兔主动脉平滑肌细胞。进行vonKossa染色以显示钙化。生化法检测细胞内外不溶性钙。放免法测定培养上清骨钙素的含量。RT PCR方法检测X型胶原mR NA的表达。结果 :2 5 羟基胆固醇 (A组 )和β 甘油磷酸盐 (B组 )分别培养的传代细胞均可见多个细胞结节 ,且细胞结节vonkossa染色阳性 ;而对照组 (C组 )无细胞结节形成 ,vonkossa染色阴性。前二者每孔不溶性钙沉积量 ,以及培养上清中骨钙素含量明显高于后者 ,且A、B两组X型胶原mRNA的表达为阳性。结论 :2 5 羟基胆固醇、β 甘油磷酸盐均可促进主动脉中膜细胞发生钙化。主动脉中膜在钙化过程中与成骨细胞相似 。Aim: To investigate the stimulation to in vitro calcification of aorta by 25 hydroxycholesterol or β glycerophosphate, and the relationship of the calcification with the secretion of osteocalcin and expression of type X collagen mRNA. Methods: Aortic medial cells were obtained by explantation. Von Kossa staining was performed to show the in vitro calcification. Insoluble calcium of cellular layer was determined by biochemical method. And osteocalcin in the media was analyzed with radioimmunoassay. RT PCR was used to dctermine the expression of type X collagen mRNA. Results: Passaged cells showed negative von Kossa after 28 days. While cells treated with 25 hydroxycholesterol or β glycerophosphate appeared many cell nodules with positive von Kossa staining, in witch much more insoluble calcium and supernatant osteocalcin were detected. Type X collagen mANA was detected in cells treated with 25 hydroxycholesterol or β glycerophosphate but not in untreated cells. Conclusion: 25 hydroxycholesterol and β glycerophosphate can accelerate the in vitro calcification of aortic media. Aortic medial cells secrete more osteocalcin and express type X collagen mRNA during calcification, which is similar to osteoblast, suggesting arterial calcification may share part of the same mechanism with the calcification of the bone.

关 键 词：动脉钙化 X型胶原 骨钙素 25-羟基胆固醇 β-甘油磷酸盐 

分 类 号：R543.12[医药卫生—心血管疾病] Q463[医药卫生—内科学]