Rogler G, Lackner K J, Schmitz G
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany.
Am J Cardiol. 1995 Jul 13;76(2):114A-116A. doi: 10.1016/s0002-9149(05)80031-1.
The effects of fluvastatin, a new fully synthetic inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, on growth and cell-cycle kinetics of porcine and human vascular smooth muscle cells (SMC) were studied by growth curves and flow cytometric determination of cell-cycle distribution. Growth curves were obtained from counting after 2, 4, 7, 9, 11, and 14 days of incubation in Dulbecco's minimum essential medium and 10% fetal calf serum (FCS). For cell-cycle phase determination, cells were synchronized in G0 by 48 hours of incubation in serum-free medium, then stimulated by incubation in 10% FCS, with or without fluvastatin. There was a concentration-dependent decrease in the proliferation of human and porcine SMC when cells were incubated in the presence of fluvastatin. The reduction in the number of cells was significant with 10(-5) M and 10(-4) M fluvastatin. The G/S-phase transition of human and porcine vascular SMC was reduced to 50% of controls by 10(-4) M fluvastatin, as revealed by cell-cycle analysis. The effects of fluvastatin on growth kinetics and cell-cycle distribution could be completely reversed by the addition of 1 mM mevalonolactone, indicating that the fluvastatin effects are due to specific inhibition of HMG-CoA reductase. The addition of low density lipoprotein as a source of cholesterol failed to support SMC growth and phase transition. Addition of squalene or cholesterol to the culture medium also failed to normalize cell growth. It is concluded that nonsterol products synthesized from mevalonate are necessary for the growth of SMC.(ABSTRACT TRUNCATED AT 250 WORDS)
通过生长曲线以及流式细胞术测定细胞周期分布，研究了新型全合成3-羟基-3-甲基戊二酰辅酶A（HMG-CoA）还原酶抑制剂氟伐他汀对猪和人血管平滑肌细胞（SMC）生长及细胞周期动力学的影响。生长曲线通过在含有10%胎牛血清（FCS）的杜尔贝科改良伊格尔培养基中孵育2、4、7、9、11和14天后计数获得。为了测定细胞周期阶段，细胞在无血清培养基中孵育48小时使其同步于G0期，然后在含有或不含有氟伐他汀的10% FCS中孵育以进行刺激。当细胞在氟伐他汀存在的情况下孵育时，人和猪SMC的增殖呈浓度依赖性下降。10^(-5) M和10^(-4) M的氟伐他汀使细胞数量显著减少。细胞周期分析显示，10^(-4) M氟伐他汀使人和猪血管SMC的G/S期转变降至对照的50%。添加1 mM甲羟戊酸内酯可完全逆转氟伐他汀对生长动力学和细胞周期分布的影响，表明氟伐他汀的作用是由于对HMG-CoA还原酶的特异性抑制。添加低密度脂蛋白作为胆固醇来源无法支持SMC生长和阶段转变。向培养基中添加角鲨烯或胆固醇也无法使细胞生长恢复正常。结论是，由甲羟戊酸合成的非甾醇产物对SMC的生长是必需的。（摘要截短于250字）
