替普瑞酮对百草枯诱导的SH-SY5Y细胞TDP43蛋白异常聚集抑制作用的研究  

INHIBITORY EFFECT OF GERANYLGERANYLACETONE ON PARAQUAT-INDUCED ABNORMAL AGGREGATION OF TDP-43 PROTEIN IN SH-SY5Y CELLS

作　　者：崔宝龙 何晓燕 姚鲁清 赵芳 周畅[1] CUI Baolong;HE Xiaoyan;YAO Luqing;ZHAO Fang;ZHOU Chang(Department of Neurology, The Affiliated Hospital of Qingdao University, Qingdao 266003, China)

机构地区：[1]青岛大学附属医院神经内科,山东青岛266003

出　　处：《精准医学杂志》2020年第3期217-221,共5页Journal of Precision Medicine

基　　金：山东省自然科学基金资助项目(ZR2014HM064)。

摘　　要：目的探讨替普瑞酮(GGA)对百草枯(PQ)诱导的SH-SY5Y氧化应激模型细胞中TAR DNA/RNA结合蛋白43(TDP43)蛋白异常聚集的影响及其相关机制。方法将SH-SY5Y细胞分为无任何处理组(A组),以100μmol/L的GGA处理SH-SY5Y细胞24 h组(B组),分别以0、20、50、100μmol/L的GGA处理氧化应激模型细胞24 h组(C^F组)。分别用MTT比色法、流式细胞仪与免疫印迹法(Western blot)检测各组SH-SY5Y细胞的存活率、凋亡率、TDP43蛋白表达水平、P62蛋白表达水平及LC3BⅡ/Ⅰ比值。为观察GGA对TDP43蛋白表达水平影响是否通过自噬发挥作用,将SH-SY5Y细胞分为无任何处理组(A组),分别以0、50、50μmol/L的GGA处理氧化应激模型细胞24 h组(G^I组),其中I组用5 mmol/L的3-MA阻断自噬,利用Western blot检测各组SH-SY5Y细胞TDP43蛋白表达水平和LC3BⅡ/Ⅰ比值。结果A^F组SH-SY5Y细胞的存活率、凋亡率、TDP43蛋白表达水平、P62蛋白表达水平及LC3BⅡ/Ⅰ比值比较差异有显著性(F=37.57~231.50,P<0.01);其中D、E、F组的细胞存活率、凋亡率、TDP43、P62蛋白表达水平与LC3BⅡ/Ⅰ比值与C组比较差异均有显著性(q=3.01~22.23,P<0.01)。3-MA阻断自噬后,A、G^I组TDP43蛋白表达水平与LC3BⅡ/Ⅰ比值比较差异有显著性(F=40.40,66.68,P<0.01);其中I组与H组TDP43蛋白表达水平比较差异有显著意义(q=6.12,P<0.01)。结论GGA能够通过增强自噬降低PQ诱导的SH-SY5Y氧化应激模型细胞中TDP43蛋白的表达水平。Objective To investigate the effect of geranylgeranylacetone(GGA)on paraquat(PQ)-induced abnormal aggregation of TAR DNA/RNA-binding protein-43(TDP-43)in SH-SY5Y cell model of oxidative stress and its related mechanism.MethodsSH-SY5Y cells were divided into non-treatment group(group A),SH-SY5Y cells treated with 100μmol/L GGA for 24 h group(group B),and SH-SY5Y cell model of oxidative stress treated with 0,20,50,or 100μmol/L GGA for 24 h group(groups C-F).MTT colorimetric assay,flow cytometry,and Western blot were used to determine the survival rate,apoptosis rate,TDP-43 and P62 protein expression levels,and LC3BⅡ/Ⅰratio of SH-SY5Y cells in each group.In order to observe whether GGA affects TDP-43 protein expression level through autophagy,SH-SY5Y cells were divided into non-treatment group(group A),and SH-SY5Y cell model of oxidative stress treated with 0,50,or 50μmol/L GGA for 24 h group(groups G-I),of which group I was treated with 5 mmol/L 3-MA to block autophagy.Western blot was used to determine the TDP-43 protein expression level and LC3BⅢ/Ⅰratio of SH-SY5Y cells in each group.Results There were significant differences in the survival rate,apo-ptosis rate,TDP-43 and P62 protein expression levels,and LC3BⅡ/Ⅰratio of SH-SY5Y cells between groups A-F(F=37.57-231.50,P<0.01);compared with group C,groups D,E,and F had significantly different cell survival rate,apoptosis rate,TDP-43 and P62 protein expression levels,and LC3BⅡ/Ⅰratio(q=3.01-22.23,P<0.01).After autophagy was blocked using 3-MA,there was a significant difference in the protein expression level of TDP43 and LC3BⅡ/Ⅰratio between groups A and G-I(F=40.40,66.68,P<0.01),among which there was a significant difference in the protein expression level of TDP-43 between groups I and H(q=6.12,P<0.01).Conclusion GGA can reduce TDP-43 protein expression level in PQ-induced oxidative stress of SH-SY5Y cells by enhancing autophagy.

关 键 词：替普瑞酮 百草枯 细胞系 肿瘤 DNA结合蛋白质类 SH-SY5Y细胞 氧化性应激 自噬 

分 类 号：R746.4[医药卫生—神经病学与精神病学]