Laboratories of Cellular Signaling and Environmental Physiology, Faculty of Integrated Arts and Sciences, The University of Tokushima, Minami-Jyosanjima 1-1, Tokushima 770-8502, Japan.
Environ Toxicol Pharmacol. 1999 Oct;7(4):267-73. doi: 10.1016/s1382-6689(99)00026-5.
Effects of tri-n-butyltin chloride (TBT) on rat thymocytes were examined by using a flow cytometer and three fluorescent dyes (annexin V-FITC, ethidium bromide and fluo-3-AM) to further characterize its cytotoxic action. TBT at concentrations of 100 nM or greater, time- and dose-dependently increased the population of annexin V-positive live cells in the cell suspension. Most of cells became to be annexin V-positive within 60 min after the start of application of 300 nM TBT. Some of annexin V-positive live cells were further stained with ethidium, indicating that some of the cells were killed, in continued presence of TBT at 300 nM or greater. When the cells were exposed to 300 nM TBT only for 15 min, the population of annexin V-positive live cells increased after removal of TBT from incubation medium. TBT-induced increase in the population of annexin V-positive live cells was partly attenuated under Ca(2+)-free condition, although that was not the case for the dead cells. TBT at 30 nM or greater increased [Ca(2+)]i in a dose-dependent manner. Triethyltin and trimethyltin even at 1 μM did not increase the [Ca(2+)]i and the population of annexin V-positive live cells. The population of annexin V-positive live cells increased as the [Ca(2+)]i was increased by ionomycin, a calcium ionophore. Results suggest an involvement of Ca(2+) in some of TBT-induced cytotoxicity.
三丁基锡氯化物（TBT）对大鼠胸腺细胞的影响，采用流式细胞仪和三种荧光染料（ Annexin V-FITC 、溴化乙锭和 fluo-3-AM ）来进一步阐明其细胞毒性作用。 TBT 在 100 nM 或更高浓度，时间和剂量依赖性地增加了在细胞悬浮液中 Annexin V 阳性活细胞的群体。大多数细胞在应用 300 nM TBT 后 60 分钟内变成 Annexin V 阳性。 Annexin V 阳性活细胞中的一些进一步用溴化乙锭染色，表明在持续存在 300 nM 或更高浓度的 TBT 时，一些细胞被杀死。当细胞仅暴露于 300 nM TBT 15 分钟时，在从孵育介质中除去 TBT 后， Annexin V 阳性活细胞的群体增加。 TBT 诱导的 Annexin V 阳性活细胞群体的增加在无 Ca 2+条件下部分减弱，尽管死细胞的情况并非如此。 TBT 在 30 nM 或更高浓度以剂量依赖性方式增加 [Ca 2+] i 。三乙基锡和三甲基锡甚至在 1 μM 时也不会增加 [Ca 2+] i 和 Annexin V 阳性活细胞的群体。当钙离子载体离子霉素增加 [Ca 2+] i 时， Annexin V 阳性活细胞的群体增加。结果表明 Ca 2+参与了 TBT 诱导的部分细胞毒性。
