红景天苷通过PINK1-Parkin 通路在MPP+诱导的SH-SY5Y细胞中维持线粒体形态和功能

目的:研究红景天苷(Salidroside,Sal)对在MPP+诱导SH-SY5Y细胞线粒体形态和功能的影响及其机制。方法:采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT)检测细胞活性,Mito Tracker Red CMXRos进行线粒体染色,四甲基罗丹明乙酯(Tetramethylrhodamine ethyl ester,TMRE)检测线粒体膜电位,Western blot检测PINK1和Parkin蛋白表达水平。结果:单纯Sal处理24 h对细胞活性、线粒体形态和MMP无影响(P0.05)。MPP+(500μM)处理SH-SY5Y细胞24 h后,与正常组比较,细胞活性、MMP水平均降低,线粒体长度减短(P0.01),并发生碎片化。Sal(25μM)预处理24 h可以显著抑制MPP+诱导的细胞活性降低(P0.01),并维持线粒体长度和增加MMP水平(P0.01)。而且,Sal(25μM)预处理24 h可以显著恢复MPP+诱导的PINK1和Parkin蛋白表达水平下降(P0.01)。结论:体外实验证实Sal可以保护MPP+诱导的SH-SY5Y细胞活性降低、线粒体形态和功能异常,而PINK1-Parkin通路可能是其机制之一,为进一步临床开发Sal治疗PD的新药提供实验依据。

Salidroside Maintains Mitochondrial Morphology and Function in MPP+induced SH-SY5Y Cells through PINK1-Parkin Pathway

Objective:This study aims to investigate the mechanismof Salidroside (Sal) protecting mitochondrial morphology andfunction in MPP+ induced SH-SY5Y cells.Methods:3- (4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assaywas used to test the cell activity; MitoTracker Red CMXRos was used to test the mitochondrial morphology; Tetramethylrhodamine ethylester (TMRE) was used to test the mitochondrial membrane potential (MMP) and Western blot method was used to detect the protein levelof PINK1 and Parkin.Results:Sal treatment of 24 h had no effect on cell activity, mitochondrial morphology and MMP (P>0.05). Whencompared with the control group, the cell activity and MMP levels were statistically decreased respectively (P<0.01), and the length ofmitochondrial was statistically shortened (P<0.01) and in fragmentation after MPP+ (500 uM) treated 24 h in SH-SY5Y cells. However,pretreatment with Sal (25 uM) 24 h could significantly inhibit MPP+ induced the decrease of cellular activity (P<0.01), and maintain mitochondriallength and increase the level of MMP (P<0.01). Moreover, pretreatment with Sal (25 uM) 24 h could significantly inhibit theMPP+ (500 uM) induced the decrease of protein level of PINK1 and Parkin (P<0.01).Conclusion:Our vitro experiments showed that Salcould protect the MPP+ induced the decrease of SH-SY5Y cell activity and shortened the length of mitochondrial and in fragmentation.PINK1-Parkin pathway might be one of the mechanisms. It was provided experimental basis for Sal as a new drug for the treatment ofPD.