Sames K, Lücht B
Department of Anatomy, University of Heidelberg.
Z Gerontol. 1991 Mar-Apr;24(2):94-7.
In vitro aging fibroblasts and porcine aortic endothelium cells were treated with a commercial soy-bean lipid emulsion and incubated in 35S-sulfate or 3H-glucosamine respectively. The glycosaminoglycans (GAGs) were characterized by enzyme digestion following proteolysis. Uronic acid concentrations were determined in fibroblast cultures. The GAG content is lower in the last possible passage than in the foregoing ones. Lipid treatment leads to an increase, especially in early passages. The incorporation of radiolabeled precursors decreases with increasing in vitro age in both cell types. By lipid treatment the heparansulfate (HS) radioactivity of fibroblasts is increased insignificantly, while the GAG radioactivity of endothelium cells and the chondroitinsulfate (CS) radioactivity of fibroblasts are decreased. The in vitro age-related reduction of the precursor incorporation is more pronounced in CS as compared to HS, whereby the HS radioactivity shows a percentual increase, especially following lipid treatment. The effect of lipids shows changes related to in vitro aging. The role of cellular GAG metabolism and lipid loads in arteriosclerosis is discussed. Even in the arterial wall different cell types may react differentially.
体外老化的成纤维细胞和猪主动脉内皮细胞用市售大豆脂质乳剂处理,并分别在35S-硫酸盐或3H-葡糖胺中孵育。蛋白水解后通过酶消化对糖胺聚糖(GAG)进行表征。测定成纤维细胞培养物中的糖醛酸浓度。在最后可能的传代中,GAG含量低于前几代。脂质处理导致增加,尤其是在早期传代中。两种细胞类型中,放射性标记前体的掺入随着体外老化而减少。通过脂质处理,成纤维细胞的硫酸乙酰肝素(HS)放射性无明显增加,而内皮细胞的GAG放射性和成纤维细胞的硫酸软骨素(CS)放射性降低。与HS相比,CS中前体掺入的体外年龄相关降低更为明显,其中HS放射性显示出百分比增加,尤其是在脂质处理后。脂质的作用显示出与体外老化相关的变化。讨论了细胞GAG代谢和脂质负荷在动脉粥样硬化中的作用。即使在动脉壁中,不同的细胞类型也可能有不同的反应。