Cellular membrane fluidity measurement by fluorescence polarization in indomethacin-induced gastric cellular injury in vitro
Cellular membrane fluidity measurement by fluorescence polarization in indomethacin-induced gastric cellular injury in vitro
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DOI：
10.1007/s00535-007-2116-y
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发表时间：
2007-12-01
影响因子：
6.3
通讯作者：
Hyodo, Ichinosuke
中科院分区：
文献类型：
--
作者：
Kaneko, Tsuyoshi;Matsui, Hirofumi;Hyodo, Ichinosuke
背景吲哚美辛的胃并发症涉及活性氧的产生，其通过细胞膜的脂质过氧化而引起胃粘膜损伤。活性氧的过氧化作用改变了细胞膜中不饱和脂肪酸的含量，从而影响膜的流动性。因此，吲哚美辛诱导的脂质过氧化反应可以通过荧光偏振（FP）法测量细胞膜流动性来检测。本研究的目的是阐明FP方法检测吲哚美辛诱导的胃细胞损伤RGM-1细胞的有用性。方法.采用常规细胞毒试验、二苯基芘膦荧光法检测脂质过氧化反应、FP法检测吲哚美辛处理的RGM-1细胞。研究了自由基清除剂和引发剂对RGM-1细胞膜流动性变化（MFC）的影响。将FP检测细胞损伤的灵敏度与DPPP荧光法和常规细胞毒性测定法进行了比较。结果吲哚美辛引起MFC的增加，通过FP测定，然后通过常规方法检测细胞毒性。MFC的增加与膜磷脂过氧化（MPP）的增加有关，但与前列腺素缺乏无关，维生素E可预防MFC和MPP的增加。结论. FP法可用于体外检测细胞损伤。
Background. Gastric complications of indomethacin involve generation of reactive oxygen species, which induce gastric mucosal injury via lipid peroxidation of cell membranes. Peroxidation by reactive oxygen species alters the amounts of unsaturated fatty acids in the cell membrane and thus affects membrane fluidity. Indomethacin-induced lipid peroxidation can thus be detected by measuring cellular membrane fluidity by the fluorescence polarization (FP) method. The aim of this study was to elucidate the usefulness of the FP method for detecting indomethacin-induced gastric cellular injury in RGM-1 cells. Methods. Indomethacin-treated RGM-1 cells were investigated by conventional cytotoxicity assay, fluorometry of diphenyl-1-pyrenylphosphine (DPPP) to detect lipid peroxidation, and FP. The effects of both a radical scavenger and an initiator on membrane fluidity change (MFC) in RGM-1 cells were examined. The sensitivity of FP in detecting cellular injury was compared with those of DPPP fluorometry and conventional cytotoxicity measurements. Results. Indomethacin caused an increase in MFC as determined by FP before cytotoxicity was detected by conventional methods. The increase in MFC was associated with increased membrane phospholipid peroxidation (MPP) but not with a prostaglandin deficiency, and the increases in both MFC and MPP were prevented by vitamin E. Conclusions. The FP method is potentially useful for detecting cellular injury in vitro.