论文部分内容阅读
目的 在体外血管钙化模型基础上探讨同型半胱氨酸 (HCY)对血管钙化的影响。方法建立牛主动脉平滑肌细胞体外钙化模型 (钙化BASMCs) ,检测HCY对细胞层钙含量、培养上清骨钙素浓度及碱性磷酸酶活性的影响 ,并检测骨钙素、骨桥蛋白及Ⅰ型胶原 (ColⅠ )mRNA表达的变化及抗氧化剂N 乙酰半胱氨酸对HCY促钙化作用的影响。结果 HCY剂量依赖性促钙化BASMCs钙沉积 ,但不促进非钙化的BASMCs钙沉积 ;N 乙酰半胱氨酸抑制HCY对钙化BASMCs钙沉积的促进作用 ,但在钙化BASMCs中单纯加入等剂量的N 乙酰半胱氨酸对钙沉积无影响 ;HCY促钙化BASMCs培养上清骨钙素含量增加 ,且使骨桥蛋白、骨钙素、ColⅠmRNA表达分别增加 56 33 %、86 48%及 1 1 0 64 % ,但对正常培养的BASMCs培养上清骨钙素含量及骨桥蛋白、骨钙素mRNA表达无影响 ,仅使ColⅠmRNA表达增加 1 0 8 33 % ;HCY不影响正常及钙化BASMCs碱性磷酸酶活性。结论 (1 )HCY是钙化的促进因子 ,而非启动因子 ;(2 )HCY的促钙化作用可能部分是通过细胞外基质途径实现的 ;(3)N 乙酰半胱氨酸阻断HCY的促钙化作用 ,间接提示氧化反应参与此过程 ;(4)HCY的促钙化作用与碱性磷酸酶活性无关
Objective To investigate the effect of homocysteine (HCY) on vascular calcification based on the in vitro vascular calcification model. Methods The in vitro calcification model of bovine aortic smooth muscle cells (calcified BASMCs) was established to detect the effect of HCY on the calcium content in the cell layer, the concentration of osteocalcin in culture supernatant and the activity of alkaline phosphatase. The levels of osteocalcin, osteopontin and Ⅰ Changes of Col Ⅰ mRNA Expression and the Effect of Antioxidant N - acetylcysteine on HCY - induced Calcification. Results HCY promoted the calcium deposition of BASMCs in a dose-dependent manner, but did not promote the calcium deposition of non-calcified BASMCs. N-acetylcysteine inhibited the promotion of HCY on calcium deposition in calcified BASMCs. However, adding equal amounts of N-acetyl Cysteine had no effect on calcium deposition. The content of osteocalcin in supernatant of HCY-promoted calcified BASMCs increased, and the expression of osteopontin, osteocalcin and ColⅠmRNA increased by 56.33%, 86.48% and 11.04%, respectively , But had no effect on osteocalcin and osteopontin, osteocalcin mRNA expression in normal cultured basal culture medium, which only increased the expression of ColⅠmRNA by 108.39%. HCY did not affect alkaline phosphatase activity in normal and calcified BASMCs . Conclusions (1) HCY is a calcification promoting factor, not a promoter; (2) calcification of HCY may be partially mediated by the extracellular matrix; (3) N-acetylcysteine blocks the calcification of HCY Role, indirectly prompted oxidation reaction involved in this process; (4) HCY calcification and alkaline phosphatase activity has nothing to do