舒尼替尼经线粒体内活性氧引起心肌细胞凋亡

摘要

OBJECTIVE To investigate the cardiac toxicity of sunitinib. METHODS H9C2 cells were exposed to sunitinib 0.1, 1 and 10 μmol· L-1 for 24, 48 and 72 h. Cell viability was determined by MTT assay. Apoptosis, the level of intracellular reactive oxygen species ( ROS),mitochondrial membrane potential (ΔΨm) and caspase 3 activity were detected by flow cytometry in the 24 h-treatment group. RESULTS Compared with normal control group, sunitinib 1 and 10 μmol·L-1 significantly decreased cells viability by 22％ and 32％ for 24 h-treatment group, by 48％ and 68％ for 48-h treatment group, and by 62％ and 82％ for 72 h-treatment group (P ＜0.05). Compared with normal control group, sunitinib 1 and 10 μmol·L-1 significanty increased the level of intracellular ROS (4.41 ±0.76 vs 8.68 ±0.74, 3.57 ± 1.45; P ＜0. 05), significantly decreased the ΔΨm ( 309 ± 6 vs 244 ± 28, 174 ± 2 ) ( P ＜ 0. 05 ), increased caspase 3 activity (0.96±0. 13 vs 59.40 ± 13. 17, 79.90 ±0.06) (P ＜0. 05), and decreased cell apoptosis [ (6.03 ±0.40) ％ vs (21.05 ± 5.55 ) ％, (59.05 ± 4.62 ) ％ ] ( P ＜ 0. 05 ). CONCLUSION Sunitinib could prompt the apoptosis of H9C2 cells through mitochondrial pathway by generating intracellular ROS.%目的:探讨舒尼替尼引起心肌细胞凋亡的作用机制.方法:H9C2细胞分别用舒尼替尼0.1,1和10μmol·L-1处理24,48,72h,MTT法测定细胞存活率;流式细胞仪分别测定处理24h细胞的凋亡、细胞内活性氧(ROS)、线粒体膜电位(△ψm)水平及胱天蛋白酶3活性.结果:与同一时间点正常对照组相比,舒尼替尼1,10 μmol·L-1处理24,48,72h后,细胞存活率分别明显下降了22%和32%(24 h);41%和68%(48 h);62%和82%(72h)(P<0.05).与正常时照组相比,舒尼替尼1,10μmol·L-1作用24 h后,心肌细胞内ROS水平显著升高(4.41±0.76 vs 8.68±0.74,3.57±1.45)(P<0.05),线粗体膜电位下降(309±6 vs 244±28,174±2)(P<0.05),胱天蛋白酶3活性升高(0.96±0.13 vs 59.40±13.17,79.90±0.06)(P<0.05)及细胞凋亡率增加[(6.03±0.40)vs(21.05±5.55)%,(59.05±4.62)%](P<0.05).结论:舒尼替尼可通过诱导心肌细胞内ROS的产生,经线粒体内途径引起心肌细胞凋亡.