柔筋止颤片通过抑制ROS通路减轻MPP^+诱导SH-SY5Y细胞凋亡实验研究

Roujin Zhichan tablets attenuates MPP^+ induced SH-SY5Y cell apoptosis by inhibiting ROS pathway

目的:研究柔筋止颤片保护由MPP^+诱导的SH-SY5Y细胞损伤的影响及其机制。方法:MTT检测细胞活性,DHE检测O2-,DCFH-DA检测ROS,Western blot检测Caspases蛋白表达水平。结果:SH-SY5Y细胞经MPP^+(500μmol/L)处理24h后细胞活性降低(P<0.05),O2^-和ROS水平均升高(P<0.05),Clevaed Caspase-3、Clevaed Caspase-8和Clevaed Caspase-9蛋白水平均增高(P<0.05)。柔筋止颤片(25nmol/L)预处理24h可以显著维持细胞活性降低(P<0.05),降低O2^-和ROS的水平(P<0.05),Clevaed Caspase-3、Clevaed Caspase-8和Clevaed Caspase-9蛋白水平下降(P<0.05)。结论:体外实验证实柔筋止颤片可减少MPP^+诱导的O2^-和ROS水平升高以减少线粒体的损伤,并减少Caspases蛋白的活化以抑制细胞凋亡程序的启动,进而保护MPP^+诱导的SH-SY5Y细胞活性降低,为进一步优化柔筋止颤片的研究提供了实验依据。

Objective:This study aims to investigate the protecting mechanism of Roujin Zhichan tablets in MPP^+ induced SH-SY5 Ycells.Methods:MTT assay was used to analyze the cell activity,DHE assay was used to analyze the level of O2^-,DCFH-DA assay was used to analyze the level of ROS,and western blot assay were used to detect the protein level of Caspases.Results:Compared with the control group,SH-SY5 Ycells treated by MPP^+(500μmol/L)for 24 hshowed a decrease in cell activity(P<0.05),an increase in the level of O2^-and ROS(P<0.05),and an increase in the protein level of Clevaed Caspase-3,Clevaed Caspase-8 and Cleved Caspase-9 proteins(P<0.05).Roujin Zhichan tablets(25 nmol/L)pretreatment for 24 hsignificantly maintained the cell activity(P<0.05),decreased the level of O2^-and ROS(P<0.05),and decreased the protein level of Clevaed Caspase-3,Clevaed Caspase-8 and Clevaed Caspase-9(P<0.05).Conclusion:The vitro experiments showed that Roujin Zhichan tablets can reduce MPP+induced elevated the level of O2^-and ROS to reduce the mitochondrial damage,and reduce the level of Clevaed-Caspases to inhibit the initiation of apoptotic process,thereby protecting SH-SY5 Ycells from MPP^+induced decreased activity.It provides experimental basis for further optimization of Roujin Zhichan tablets research.