Caspase-3在柔红霉素诱导的人视网膜色素上皮细胞凋亡中作用

Caspase-3 activity during daunorubicin induced human retinal pigment epithelium cell apoptosis

目的 探讨天冬酰胺特异酶切的半胱氨酸蛋白酶(caspase)成员 caspase- 3在柔红霉素诱导的人视网膜色素上皮 (RPE)细胞凋亡中的变化 .方法 原代培养的人 RPE细胞经 2 0 0μg· L- 1 柔红霉素诱导 8,2 4和 36 h后 ,按照 caspase- 3荧光检测试剂盒说明处理细胞 ,再通过荧光比色法间接检测细胞碎片中 caspase- 3的活性 .结果 正常 caspase- 3的活性为(0 .0 94± 0 .0 0 5 ) nmol AFC,8h后增加为 (0 .44 6± 0 .0 2 9)nm ol AFC,2 4h后为 (0 .75 4± 0 .0 5 6 ) nmol AFC,36 h又下降到 (0 .486± 0 .0 33) nmol AFC,但都高于正常水平 (P<0 .0 1) .加入特异性四肽抑制物 DEVD- CHO1μL 后活性为 (0 .0 40±0 .0 0 3) nm ol AFC,显著低于正常 (P<0 .0 1) .结论 Caspase-3参与了柔红霉素诱导的人视网膜色素上皮细胞的凋亡 ,柔红霉素可使其活性增加 .

AIM To study the function of caspase-3 activity during daunorubicin induced human retinal pigment epithelium cell apoptosis. METHODS After being treated with 200 μmolL -1 daunorubicin for 8, 24 and 36 h, the primary cultured human retinal pigment epithelium cells were treated by following the protocol of a specific assay kit, then the shift of fluorescence emission was detected to determine the activity of caspase-3 indirectly. RESULTS The mean activity of the normal group was (0.094±0.005) nmol AFC, and those of induced groups were (0.446±0.029) nmol AFC,(0.754± 0.056) nmol AFC, (0.486±0.033) nmol AFC,at 8, 24 and 36 h respectively. They were all higher than those of normal group ( P <0.01) The four peptides inhibitor-DEVD-CHO decreased the activity to (0.040±0.003) nmol AFC ( P < 0.01). CONCLUSION Caspase-3 is involved in daunorubicin induced human retinal pigment epithelium cell apoptosis, and daunorubicin could raise the activity of caspase-3.