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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(P>0.05)。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(P<0.01),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(P<0.01),并维持线粒体长度和增加MMP水平(P<0.01)。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(P<0.01)。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。
AIM: To investigate the effects of Salidroside (Sal) on the mitochondria morphology and function in SH-SY5Y cells induced by MPP + and its mechanism. Methods: The reaction of 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl Cell viability was measured by MTT assay. Mitochondrial staining was performed with MitoTracker Red CMXRos. Mitochondrial membrane potential was measured by TMRE. Western blot was used to detect the expression of PINK1 and Parkin. . Results: Sal alone treatment for 24 h had no effect on cell activity, mitochondria morphology and MMP (P> 0.05). After treated with MPP + (500μM) for 24 h, the cell activity and MMP level decreased and the length of mitochondria decreased (P <0.01) and fragmented compared with the normal group. Pretreatment with Sal (25 μM) for 24 h significantly inhibited MPP + -induced decrease in cell viability (P <0.01), maintained mitochondrial length and increased MMP levels (P <0.01). Moreover, pretreatment with Sal (25μM) for 24 h remarkably restored the decrease of PINK1 and Parkin protein expression induced by MPP + (P <0.01). Conclusions: In vitro experiments showed that Sal can protect MPP + -induced SH-SY5Y cells from decreasing in activity and mitochondrial morphology and function. PINK1-Parkin pathway may be one of its mechanisms, providing experimental basis for further clinical development of new drugs for treatment of PD.