培养的人二倍体细胞中与年龄相关的氢过氧化磷脂酰胆碱积累及其α-生育酚预防作用

Age-related accumulation of phosphatidylcholine hydroperoxide in cultured human diploid cells and its prevention by alpha-tocopherol.

作者信息

Suzuki T, Miyazawa T, Fujimoto K, Otsuka M, Tsutsumi M

机构信息

Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoku University, Sendai, Japan.

出版信息

Lipids. 1993 Aug;28(8):775-8. doi: 10.1007/BF02536004.

DOI:10.1007/BF02536004

PMID:8377593

Abstract

The levels of phosphatidylcholine hydroperoxide in serially cultured human fetal diploid fibroblasts at various population doubling levels were determined by high-performance liquid chromatography combined with chemiluminescence detection. This methodology utilizes a mixture of cytochrome c and luminol as post-column hydroperoxide group specific luminescent reagents. The cellular hydroperoxide content increased with age from 0.34 to 27.72 pmol/10(6) cells. At the end of the cells' in vitro lifespan (51st population doubling level), the hydroperoxide content per 10(6) cells reached about 80 times the level found in cells of the 20th population doubling level. Supplementation of exogenous alpha-tocopherol to the culture medium prevented hydroperoxide accumulation, but did not extend the lifespan in vitro. The results indicate that substantial intracellular phospholipid hydroperoxide accumulation occurred in the course of aging of human fetal diploid fibroblasts.

摘要

采用高效液相色谱结合化学发光检测法，测定了不同群体倍增水平下连续培养的人胎儿二倍体成纤维细胞中磷脂酰胆碱氢过氧化物的水平。该方法利用细胞色素c和鲁米诺的混合物作为柱后氢过氧化物基团特异性发光试剂。细胞氢过氧化物含量随年龄增长从0.34 pmol/10(6) 个细胞增加到27.72 pmol/10(6) 个细胞。在细胞体外寿命结束时（第51个群体倍增水平），每10(6) 个细胞的氢过氧化物含量达到第20个群体倍增水平细胞中含量的约80倍。向培养基中添加外源性α-生育酚可防止氢过氧化物积累，但并未延长体外寿命。结果表明，人胎儿二倍体成纤维细胞衰老过程中发生了大量细胞内磷脂氢过氧化物积累。