甘油醛-3-磷酸脱氢酶核转位与高糖应激诱导的H9c2细胞凋亡密切相关

High glucose-induced apoptosis of H9c2 cells is closely related to GAPDH nuclear translocation

目的:探讨高糖应激是否引起心肌细胞中甘油醛-3-磷酸脱氢酶(GAPDH)核转位以及GAPDH核转位是否与细胞凋亡有关。方法:高糖培养H9c2细胞48 h后,收集细胞进行细胞凋亡的测定,用DCFH-DA荧光探针测定细胞中活性氧簇(ROS)的水平,用Western Blotting方法测定细胞质和细胞核中GAPDH以及细胞中诱导型一氧化氮合酶(iNOS)的表达,用细胞免疫荧光的方法测定GAPDH的核转位情况。结果:与低糖培养组相比,高糖培养组H9c2细胞凋亡率增高,ROS生成和iNOS表达增多,GAPDH由细胞质向细胞核转位;而线粒体呼吸链复合体I抑制剂鱼藤酮和iNOS抑制剂1400W均阻断GAP-DH的转位,并减少高糖诱导的H9c2细胞的凋亡。结论:高糖诱导H9c2细胞中GAPDH由细胞质向细胞核转位;鱼藤酮和1400W可抑制高糖Flavonoids诱导的细胞凋亡,这一作用与抑制GAPDH核转位有关。

AIM： To investigate whether high glucose induces GAPDH nuclear translocation and whether the GAPDH nuclear translocation is related to high glucose-induced apoptosis of H9c2 cells. METHODS： H9c2 cells were cultured for 48 h in high glucose condition,and then cells were collected for apoptosis determination.The generation of reactive oxygen species（ROS） was determined.The expression of GAPDH and iNOS was measured by western blotting.The translocation of GAPDH was determined by immunofluorescence.RESULTS：High glucose induced apoptosis of H9c2 cells.High glucose induced oxidative stress in H9c2 cells and translocation of GAPDH to nucleus.Rotenone and iNOS inhibitor 1400W protected H9c2 cells from death by inhibiting GAPDH translocation to nucleus.CONCLUSION： High glucose induces GAPDH translocation from cytoplasm to nucleus.The inhibition of high glucose-induced apoptosis of H9c2 cells is closely related to GAPDH nuclear translocation.