Carpenter-Deyo L, Duimstra J R, Hedstrom O, Reed D J
Department of Biochemistry and Biophysics, Oregon State University, Corvallis.
J Pharmacol Exp Ther. 1991 Aug;258(2):739-46.
To determine whether incubation for several hours with intracellular Ca++ indicators caused toxicity to freshly isolated hepatocytes from rats, cells were incubated under 95% O2-5% CO2 in medium containing 2 mM Ca++ and the acetoxymethyl (AM) esters of Quin 2, Indo 1, Fluo 3, 5,5'-Dimethyl BAPTA, 4,4'-Difluoro BAPTA or Fura 2 for up to 5 hr. Quin 2-AM and Indo 1-AM (2.5 microM) induced lipid peroxidation in the cells after 1 or 3 hr of treatment, respectively. Additional experiments with Quin 2-AM (25 microM) revealed that it also caused lactate dehydrogenase leakage, cell blebbing and vitamin E loss in cells, but did not affect reduced glutathione or intracellular Ca++ content. The ability of Quin 2-AM to cause toxicity was dependent on the amount of Quin 2 which was present in the cell. Ca++ appeared to be involved in the mechanism of Quin 2-AM toxicity, for modulation of the extracellular Ca++ concentration partially inhibited lipid peroxidation, vitamin E loss, cell blebbing and lactate dehydrogenase leakage.
为了确定用细胞内钙离子指示剂孵育数小时是否会对新鲜分离的大鼠肝细胞产生毒性，将细胞在含2 mM钙离子以及喹啉2、indo - 1、Fluo - 3、5,5'-二甲基BAPTA、4,4'-二氟BAPTA或Fura - 2的乙酰氧甲基（AM）酯的培养基中，于95%氧气-5%二氧化碳条件下孵育长达5小时。喹啉2 - AM和indo - 1 - AM（2.5 microM）分别在处理1小时或3小时后诱导细胞内脂质过氧化。用25 microM喹啉2 - AM进行的额外实验表明，它还会导致细胞内乳酸脱氢酶泄漏、细胞起泡和维生素E损失，但不影响还原型谷胱甘肽或细胞内钙离子含量。喹啉2 - AM产生毒性的能力取决于细胞内喹啉2的含量。钙离子似乎参与了喹啉2 - AM的毒性机制，因为调节细胞外钙离子浓度可部分抑制脂质过氧化、维生素E损失、细胞起泡和乳酸脱氢酶泄漏。
