何首乌提取物对MPP^+诱导的SH-SY5Y细胞损伤的保护作用

Protective effect of extract of Polygonum multiflorum on SH-SY5Y cells injured by MPP^+

目的研究何首乌提取物对1-甲基-4-苯基-吡啶离子(MPP+)诱导SH-SY5Y细胞损伤的保护作用。方法体外培养SH-SY5Y细胞,建立MPP+致SH-SY5Y细胞损伤模型,实验分为对照组、MPP+损伤模型组、何首乌提取物干预组。干预组在MPP+损伤的基础上,给予不同浓度何首乌提取物(终质量浓度5、25、100 mg/L)。MTT比色法检测细胞活力;Hoechst33258染色和白光下观察细胞形态变化,比色法检测细胞培养上清液中乳酸脱氢酶(LDH)及细胞中丙二醛(MDA)含量;通过荧光强度观察活性氧(ROS)的含量和线粒体膜电位的变化。结果与对照组相比,模型组中细胞活性显著降低,Hoechst33258染色和白光下可见细胞胞体变小、核皱缩、碎裂有明显的颗粒状和固缩状荧光,并且细胞上清液中LDH泄漏明显增多、细胞中MDA含量和ROS显著升高而线粒体膜电位显著降低(P<0.05)。与模型组比较,预先4 h给予不同浓度的何首乌提取物(5、25、100 mg/L)能明显改善SH-SY5Y细胞的活性,降低细胞产生的LDH、MDA含量,并恢复线粒体膜高能电位,且对ROS有明显的抑制效果(P<0.01)。结论何首乌提取物对MPP+诱导的SH-SY5Y细胞损伤具有显著的保护作用,其保护作用与其抗氧化应激作用有关。

Objective To investigate the protective effect of extract of Polygonum multiflorum（ EPM） on SH-SY5 Y cells injured with MPP＋. Methods SH-SY5 Y cells were exposed to various doses of EPM for 4h,and then treated with 0.5 mmol / L MPP＋for 48 h. The cell viability was detected with MTT assay,and cell morphology was observed with microscope and Hoechst 33258 staining. The level of malondialdehyde（ MDA） and activity of lactate dehydrogenase（ LDH） were measured with UV spectrophotometer. Reactive oxygen species（ ROS） and the mitochondrial membrane potential were detected with fluorescence microscope. Results Compared with the control group,the viability of SH-SY5 Y cells was decreased to44.7%and the shrunk cell body and nuclear condensation were also observed in MPP＋-treated group.Meantime,the activity of LDH,level of MDA and ROS were significantly increased,but the mitochondrial membrane potential was reduced. However,pretreatment with EPM（ 5,25 and 100 mg / L） prior to MPP＋administration rescued the cell viability,decreased the levels of LDH,MDA and ROS and restored mitochondrial membrane potential in SH-SY5 Y cells. Conclusion EPM can protect the SH-SY5 Y cells against the damage induced by MPP＋,which may be involved in anti-oxidant activity.