摘要
目的观察异菝葜皂苷元(SMI)对氧化应激损伤人神经母细胞瘤株(SH-SY5Y)细胞的保护作用,并初步探讨其分子机制。方法将SH-SY5Y细胞用不同浓度(0.01、0.1、1μmol/L)SMI或等体积DMSO预处理24 h,然后加入600μmol/L过氧化氢(H2O2)处理。显微镜下观察SH-SY5Y细胞形态学变化,噻唑蓝(MTT)法检测细胞存活率,流式细胞仪检测胞内活性氧(ROS)水平,Western blotting检测p-Akt/Akt和HSP70的表达,并检测PI3K抑制剂LY294002对HSP70表达的影响。结果 SMI预处理浓度依赖地改善H2O2损伤SH-SY5Y细胞的形态,提高损伤细胞存活率,其中0.1、1μmol/L SMI预处理细胞存活率显著升高(P<0.05,P<0.001)。1μmol/L SMI显著抑制H2O2诱导的ROS升高(P<0.05)。H2O2诱导p-Akt/Akt和HSP70表达,但SMI预处理进一步提高p-Akt/Akt和HSP70表达水平(P<0.01,P<0.05),且PI3K抑制剂能阻断HSP70表达的升高(P<0.001)。结论 SMI对H2O2诱导的氧化应激损伤SH-SY5Y细胞具有明显的保护作用,其作用机制可能与降低胞内ROS水平及高度激活PI3K/Akt/HSP70信号通路有关。
Objective To observe the protective effect of smilagenin （SMI） on oxidative stress damage of human neuroblastoma cells （SH-SY5Y） and preliminarily investigate the possible molecular mechanisms. Methods SH-SYSY cells were pretreated with different concentrations （0.01, 0.1, and 1 μmol/L） of SMI or an equal volume of DMSO for 24 h and then added H2O2 of 600 μmol/L. Morphological changes of SH-SYSY cells were observed under the inverted microscope. Cell viability was detected by the MTT method. Reactive oxygen species （ROS） was detected by the flow cytometer. The expressions of p-Akt/Akt and HSP70 and the effect of PI3K inhibitor LY294002 on the expression of HSP70 were detected by the Western blotting. Results SMI pretreatment improved the morphology and cell viability of SH-SY5Y cells damaged by H2O2 in a dose dependent manner. The cell viability significantly increased after being treated by SMI of 0.1 and 1 μmol/L （P〈 0.05, P〈0. 001）. Pretreatment by SMI of 1 μmol/L significandy inhibited the increase of ROS induced by H2O2（P 〈0.05）. H2O2 induced the expression of p-Akt/Akt and HSP70 and SMI pretreatment further increased expressions of p-Akt/Akt and HSP70 （P〈0.01, P〈0.05）. PI3K inhibitor could block the increase of HSP70 expression （P 〈0. 001）. Conclusion SMI can significantly protect SH-SY5Y cells against H2O2- induced oxidative stress damage, which may be relevant to down-regulating the intracellular ROS level and highly activating the PI3K/Akt/HSP70 sinaline nathwav.
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2015年第5期631-636,共6页
Journal of Shanghai Jiao tong University:Medical Science
基金
国家自然科学基金(81001426)
上海市教委基金(12YZ038)
教育部归国学者基金~~
