Kanatate T, Nagao N, Sugimoto M, Kageyama K, Fujimoto T, Miwa N
Department of Cell Biochemistry, Hiroshima Prefectural University School of BioScience, Japan.
Cell Mol Biol Res. 1995;41(6):561-7.
Human or mouse epidermal keratinocytes NHEK or Pam212 was less susceptible to ultraviolet (UV)-B irradiation than mouse neuroblastoma NAs1 cells in culture, undergoing apoptosis-like cell death as shown by cell fragmentation and cell membrane integrity disruption. UV susceptibility was appreciably reduced by the reactive oxygen species (ROS)-scavenger L-ascorbic acid-2-phosphate (Asc2P) endowed with long-lasting functions but not by L-ascorbic acid (Asc) for each cell type. DehydroAsc reduced UV susceptibility of Pam212 or NAs1 established cell lines but not of normal diploid NHEK cells destined to be thereafter submitted to cellular senescence. The susceptibility reduction may not be ascribed to extracellular Asc2P or DehAsc, which was removed by aspirating and/or rinsing upon irradiation after the intracellular channelyzer analysis and dead cell-specific DNA-intercalator ethidium homodimer/fluorometry, respectively. Thus, the three cell types differed in UV susceptibility partly because of their different ROS-scavenging abilities, which may be potently promoted by Asc2P or dehydroAsc but not Asc.
在培养过程中,人或小鼠的表皮角质形成细胞(NHEK或Pam212)比小鼠神经母细胞瘤NAs1细胞对紫外线(UV)-B照射的敏感性更低,表现为细胞破碎和细胞膜完整性破坏的凋亡样细胞死亡。对于每种细胞类型,具有持久功能的活性氧(ROS)清除剂L-抗坏血酸-2-磷酸酯(Asc2P)可明显降低UV敏感性,而L-抗坏血酸(Asc)则不能。脱氢抗坏血酸(DehydroAsc)降低了Pam212或NAs1建立的细胞系的UV敏感性,但对注定要进入细胞衰老的正常二倍体NHEK细胞则没有作用。敏感性降低可能不归因于细胞外的Asc2P或DehAsc,在分别进行细胞内通道分析和死细胞特异性DNA嵌入剂乙锭同二聚体/荧光测定后,通过照射后抽吸和/或冲洗可将其去除。因此,这三种细胞类型在UV敏感性上存在差异,部分原因是它们具有不同的ROS清除能力,Asc2P或脱氢抗坏血酸可有效促进这种能力,而Asc则不能。