羟基红花黄色素A对MPP^+诱导的SH-SY5Y细胞凋亡的保护作用

Protective effects of hydroxysafflor yellow A against MPP^+-induced apoptosis in SH-SY5Y cells

目的观察羟基红花黄色素A(HSYA)对MPP+诱导的SH-SY5Y细胞凋亡的保护作用,并探讨其作用机制。方法采用MTT法检测细胞存活率,流式细胞术检测细胞凋亡率;二乙酸二氯荧光素盐(DCFH-DA)法检测细胞内活性氧(ROS)含量;罗丹明123(Rhodamine123)染色法检测细胞线粒体膜电位;Western blot法检测Cyt-C、Bcl-2、Bax、Caspase-3和Caspase-9的表达。结果 1mmol/L MPP+处理SH-SY5Y细胞48h后,细胞存活率显著降低,并诱导细胞产生凋亡,凋亡率达(38.6±1.8)%。而HSYA(15、60、120μmol/L)预处理1h,可明显增加SH-SY5Y细胞存活率(P<0.001),并能有效抑制MPP+诱导的细胞调亡(P<0.01);可明显抑制MPP+诱导的细胞内ROS的增加(P<0.001);也能有效抑制MPP+诱导的细胞线粒体膜电位的降低(P<0.01)。120μmol/L HSYA预处理能显著性增加Bcl-2/Bax比值(P<0.01);能够明显抑制MPP+诱导的胞质Cyt-C的释放(P<0.05)和Caspase-3、Caspase-9的高表达(P<0.05,P<0.05)。结论 HSYA预处理能明显提高MPP+诱导的SH-SY5Y细胞损伤的细胞存活率,可有效抑制细胞调亡的发生,其机制可能与抑制细胞线粒体凋亡途径有关。

Objective To investigate the neuroprotective effect of hydroxysafflor yellow A （HSYA） on MPP＋-induced apoptosis in SH-SY5Y cells and its related mechanisms.Methods Cell viability was analyzed using MTT assay.Cell apoptosis was measured by flow cytometry.The level of reactive oxygen species （ROS） within the cells was analyzed by fluorometric probe DCFH-DA assay and mitochondrial transmembrane potential was revealed by rhodamine123 staining.The expressions of cytosolic Cyt-C,Bcl-2,Bax,Caspase-3 and Caspase-9 were analyzed by Western blot.Results The cell viability was significantly decreased by 1 mmol/L MPP＋,and the percentage of apoptosis increased to （38.6 ± 1.8）％,but was reversed by HSYA pretreatment of different concentrations （15,60 and 120μmol/L） （P＜0.001,P＜0.01,respectively）.Pretreatment of HSYA reversed the increase of ROS,cytosolic Cyt-C,Caspase-3 and Caspase-9 （P＜0.001,P＜0.05,P＜0.05,P＜0.05,respectively）,decrease of mitochondrial transmembrane potential and Bcl-2/Bax ratio induced by MPP＋ in SH-SY5Y cells （P＜0.01,P＜0.01,respectively）.Conclusion HSYA pretreatment can increase cell viability to prevent MPP＋-induced cell apoptosis.The mechanism may be correlated to the inhibition of mitochondrial apoptotic pathway.