摘要
目的研究硫化氢(H2S)是否通过调控p38丝裂原活化蛋白激酶(MAPK)通路保护H9c2心肌细胞对抗阿霉素(DOX)引起的损伤。方法应用DOX处理心肌细胞建立心肌细胞损伤模型。为观察H2S的保护作用,在DOX处理心肌细胞前,应用400μmol/L硫氢化钠(NaHS,为H2S的供体)预处理细胞30 min。Western blot法测定p38MAPK蛋白的表达水平;应用细胞计数试剂盒8(CCK-8)比色法测定细胞存活率;Hoechst 33258核染色法观察细胞凋亡的形态学改变;双氯荧光素(DCFH-DA)染色荧光显微镜照相测定细胞内活性氧(ROS)水平。结果在15~60min的时间范围内,5μmol/L DOX呈时间依赖性地上调心肌细胞磷酸化p38MAPK的表达水平;在DOX作用心肌细胞前,400μmol/L NaHS预处理30 min能明显地抑制DOX对p-p38MAPK表达的上调作用,并能显著地阻断DOX引起的心肌细胞损伤,使细胞存活率升高、凋亡细胞数量和ROS生成均减少;与NaHS的保护作用相似,p38MAPK的抑制剂SB203580(3μmol/L)预处理60 min能保护H9c2心肌细胞对抗DOX引起的损伤。结论 p38MAPK通路参与DOX对心肌细胞的损伤作用;H2S可通过抑制p38MAPK通路保护心肌细胞对抗DOX诱导的损伤。
Aim To explore whether hydrogen sulfide （H2S） protects H9c2 cells against doxorubicin-induced injury by modulating p38 mitogen-activated protein kinase（MAPK） pathway. Methods H9c2 ceils were treated with doxorubicin（DOX） to establish the model of cardiac cell injury; In order to explore the protective effect of H2S, cells were pretreated with 400 Ixmol/L NariS （a donor of H2S）for 30 min before exposure to DOX. The expression level of p38MAPK protein was tested by Western blot; Cell viability was measured by cell counter kit-8 （CCK-8） ; Morphological changes of apoptotic cells were detected by Hoechst 33258 staining; Intracellular level of reactive oxygen species （ROS） was measured by DCFH-DA staining and photofluorography. Results At range of 15 to 60 min, DOX at 5 p^mol/L time-dependently upregulated expression level of phosphorylated（p） p38MAPK. Pretreatment with 400 Ixmol/L NariS for 30 min before exposure of H9c2 cells to DOX not only obviously inhibited upregulation of p-p38MAPK expression induced by DOX, but also significantly blocked DOX-induced cardiomyocyte injuries, as evidenced by an increase in cell viability, decreases in amount of apoptotic cells and intracellular ROS generation. Similar to the cardioprotective effect of NariS, pretreatment with SB203580 （ an inhibitor of p38MAPK） for 60 min also protected H9c2 cardiac cells against DOX-induced injury. Conclusion p38MAPK pathway participates in DOX-induced cardiomyocyte damage; H2S may protect cardiomyocyte against DOX-induced injury by inhibiting p38MAPK pathway.
出处
《中国动脉硬化杂志》
CAS
CSCD
北大核心
2013年第8期690-694,共5页
Chinese Journal of Arteriosclerosis
基金
广东省科技计划基金项目(2010B080701035和2012B031800358)
关键词
硫化氢
P38丝裂原活化蛋白激酶
阿霉素
心肌细胞
活性氧
细胞凋亡
Hydrogen Sulfide
p38 Mitogen-Activated Kinase
Doxorubicin
Cardiomyocyte
Reactive Oxy-gen Species
Apoptosis
