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为研究主动脉中膜细胞体外钙化情况以及 2 5 -羟基胆固醇对此过程的促进作用 ,采用贴块法分离培养主动脉中膜细胞 ,并进行vonKossa染色以显示钙化 ,生物化学检测细胞不溶性钙 ,放射免疫法测定培养上清液中骨钙素含量。发现原代细胞有两种生长表现 :一种细胞平行生长 ,不形成结节 ,vonKossa染色阴性 ;另一种易形成结节 ,vonKossa染色阳性。对照组传代细胞培养 2 8天无结节形成 ,vonKossa染色阴性。而实验组 ,即 2 5 -羟基胆固醇处理组有细胞结节形成 ,vonKossa染色阳性 ,培养上清液中骨钙素增多。由此可见体外培养的主动脉中膜细胞有两种亚型 ,一种为平滑肌细胞的表型 ;另一种为微血管周细胞型细胞 ,能使其细胞外基质钙化。 2 5 -羟基胆固醇可以促进这一体外钙化过程。周细胞型细胞钙化过程中骨钙素合成增多 ,推测骨钙素可能参与了主动脉的钙化。
In order to study the in vitro calcification of aortic mesangial cells and the promotion of 25-hydroxycholesterol on this process, the aortic mesangial cells were isolated and cultured by patch method and stained with von Kossa to show calcification. The biochemical detection of insoluble calcium, Determination of osteocalcin in culture supernatant by radioimmunoassay. Primary cells were found to have two growth manifestations: one grew in parallel and did not form nodules, von Kossa stained negative; the other formed nodules von Kossa stained positive. The control group of passaged cells cultured 28 days without nodules, vonKossa staining negative. The experimental group, that is, 2 5 - hydroxycholesterol treatment group formed nodules, vonKossa staining positive, osteocalcin in culture supernatants increased. This shows that there are two subtypes of aortic intima cultured in vitro, one is the phenotype of smooth muscle cells; the other is microvascular pericytes, calcification of the extracellular matrix. 2 5 - hydroxycholesterol can promote this in vitro calcification process. Osteocalcin synthesis increased during calcification of pericytes, suggesting that osteocalcin may be involved in aortic calcification.