Ndfip1对MPP^+诱导的SH-SY5Y细胞损伤的保护作用

Neuro-protection effect of Ndfip1 on MPP^+ induced SH-SY5Y cellular damage

目的探讨Ndfip1对MPP+(1-甲基-4-苯基-1,2,3,6-四氢吡啶)诱导的帕金森病(Pkarkinson's disease,PD)细胞模型(人神经母细胞瘤SH-SY5Y细胞)的保护作用。方法利用已建立好的Ndfip1内源性表达(野生型,WT-SY5Y)、过表达(4HT诱导,SY5Y-N)、显性负表达(显性负载体,SY5Y-D)及抑制表达(Ndfip1 shRNA载体瞬时转染,SY5Y-I)四种不同SH-SY5Y细胞模型,采用MPP+诱导上述细胞形成PD细胞模型,利用MTT法在酶联免疫检测仪上测量OD490吸光值以计算细胞的存活率,采用Annexin V凋亡检测试剂盒和流式细胞仪测定细胞的凋亡率。结果 0.3 mmol/L浓度的MPP+可降低WT-SY5Y细胞存活率,增加细胞凋亡率,其他细胞模型的细胞存活率及细胞凋亡率也与此类似;当加入4HT时,SY5Y-N细胞中因Ndfip1大量表达可明显增加细胞存活率,抑制细胞凋亡率,与WT-SY5Y相比,差异有统计学意义(P<0.05);而此时SY5Y-D和SY5Y-I细胞中因Ndfip1的表达被抑制不能明显增加细胞存活率,降低细胞死亡率(P>0.05)。结论 Ndfip1对MPP+诱导的PD细胞模型具有明显的神经保护作用及抗凋亡作用,研究结果有助于对PD发病机制的研究,为PD的临床治疗开辟了新的思路。

Objective To investigate the neuro-protection of Ndfip1 on 1-methyl-4-phenyl-5-4 hydrogen pyridine（MPP＋） induced Pkarkinson＇s disease（PD） cell model（human neuroblastoma SH-SY5Y）.Methods 4 kinds of different SH-SY5Y cell models（WT-SY5Y for endogenous expression of Ndfip1,SY5Y-N for over-expression of Ndfip1,SY5Y-D for dominant negative Ndfip1 expression and SY5Y-I for inhibition of Ndfip1 expression） were already set up,and MPP＋ was used to induce the cell models to form PD model.OD490 absorbance was measured on the microplate reader by the MTT method to calculate the cell survival rate,while the apoptotic rate was detected by flow cytometry according to Annexin V apoptosis detection kit.Results 0.3 mmol/L MPP＋ could decrease the survival rate of WT-SY5Y and increase the apoptotic rate of WT-SY5Y.The similar viability and apoptotic rates could be measured in other model cells after adding with 0.3 mmol/L MPP＋.After adding 4HT,the over-expression of Ndfip1 in SY5Y-N cells had the function to increase the viability and decrease the apoptotic rate,which showed significant differences compared with WT-SY5Y（P〈0.05）.At the same time,inhibition of Ndfip1 expression in SY5Y-D and SY5Y-I cells could not in crease the viability and decrease the apoptotic rate dominant significantly（P〈0.05）.Conclusion Ndfip1 has distinct functions of neuro-protection and anti-apoptosis on MPP＋ induced PD model cells.The results will contribute to the study of the pathogenesis of PD and will help to open up a new train of thought for clinical treatment of PD.