羟丙基-β-环糊精通过加重自噬障碍降低稳转hSOD1G93A的NSC34细胞的活力

Hydroxypropyl-β-cyclodextrin decreases the viability of NSC34 cells with hSODIG93A stably transfected by aggravating autophagic flux impairment

目的观察羟丙基-β-环糊精(HpβCD)对稳转hSOD1G93A的NSC34细胞活力的影响并探讨可能的机制。方法应用HpBCD分别干预对照组(稳转空质粒的NSC34细胞)和实验组(稳转hSOD1G93A的NSC34细胞),通过CCK-8试剂盒检测各组细胞活力,通过透射电镜观察细胞自噬结构,应用Western blotting检测human SOD1、LC3Ⅱ/Ⅰ、P62的蛋白表达水平。结果HpβCD干预后,稳转hSOD1G93A的NSC34细胞活力下降,细胞中humanSOD1含量及自噬结构明显增多,LC3Ⅱ/Ⅰ、P62蛋白的表达水平也显著增加。结论HpβCD降低稳转hSOD1G93A的NSC34细胞活力,这种细胞毒性作用可能是由于进一步加重了稳转hSOD1G93A的NSC34细胞的自噬障碍。

Objective To observe the effect of hydroxypropyl-β-cyclodextrin(HpβCD)on cell viability of NSC34 cells with hSOD1G93A stably transfected and explore the possible mechanism.Methods HpβCD were used to incubate two groups:NSC34 cells with empty plasmid stably transfected and NSC34 cells with hSODlG93A stably transfected.Cell viability was determine by CCK-8,changes in the autophagic vacuoles were observed by transmission electron microscopy,and the protein expression levels of human SOD1,LC3Ⅱ/Ⅰand P62 proteins were detected by western blotting.Results HpβCD decreased the viability of NSC34 cells with hSOD1G93A stably transfected,the human SOD1 and autophagic vacuoles increased,and the expression levels of LC3Ⅱ/Ⅰand P62 were higher in the treated groups than in the untreated groups.Conclusion Hpβ3CD decreased the viability of NSC34 cells with hSODlG93A stably transfected,and these cytotoxic effects were related to aggravate autophagic flux impairment.