ABT-737诱导秀丽线虫生殖腺细胞凋亡及机制研究

The mechanism of ABT-737-induced apoptosis of gonadal cells in Caenorhabditis elegans

目的探讨ABT-737在秀丽线虫活体水平对生殖腺细胞凋亡的作用及机制研究。方法荧光染料法检测不同浓度处理下野生型秀丽线虫生殖腺细胞凋亡的变化,同时利用检测活性氧自由基(ROS)的荧光线虫品系和多种基因突变线虫品系明确ABT-737在活体水平诱导凋亡的机制。结果在浓度为0、10、20μmol/L的ABT-737处理后,野生型秀丽线虫凋亡细胞数目从(2.31±0.06)个增加至(4.02±0.09)、(4.40±0.08)个。然而,在有丝分裂原活化蛋白激酶通路(MAPK/JNK通路,mek-1/jnk-1)以及DNA损伤应答途径(egl-1、hus-1、cep-1)基因突变品系中凋亡未有变化。此外,ABT-737导致SOD3荧光品系秀丽线虫的荧光强度增加,即线虫ROS水平上升。结论ABT-737在活体动物水平以剂量依赖的方式诱导秀丽线虫生殖腺细胞凋亡;氧化应激、DNA损伤应答通路及MAPK/JNK信号通路参与凋亡过程的发生。

Objective To investigate the effect and mechanism of ABT-737-mediated apoptosis in Caenorhabditis elegans(C.elegans)of living animals.Methods Fluorescent dye method was used to detect the apoptosis of germ cell in wild-type C.elegans under different concentrations of treatment.Meanwhile,reactive oxygen species(ROS)fluorescent strain together with different gene mutant strains of C.elegans were used to elucidate the mechanism of ABT-737-induced apoptosis.Results After treated with ABT-737 at concentrations of 0,10,and 20μmol/L,the number of apoptotic cells in wild-type C.elegans increased from 2.31±0.06 to 4.02±0.09 and4.40±0.08.However,there was no change of apoptosis in C.elegans lacking the expression of the mitogen-activated protein kinase pathway(MAPK/JNK pathway,mek-1/jnk-1)and DNA damage response pathway(egl-1,hus-1,cep-1)genes.In addition,ABT-737 increased the fluorescence intensity in the SOD3 fluorescent strain,indicating the ROS level was increased in C.elegans.Conclusion ABT-737 induces apoptosis of C.elegans germ cells in a dose-dependent manner at the level of live animals.Oxidative stress,DNA damage response pathway and MAPK/JNK signaling pathway are involved in ABT737-induced apoptotic signaling.