miR-17-5p调控STAT1表达对Aβ_(1-42)诱导SH-SY5Y细胞损伤的影响

Impact of miR⁃17⁃5p regulating STAT1 expression on Aβ_(1⁃42)⁃induced SH⁃SY5Y cell injury

目的探讨miR-17-5p过表达调控信号转导子与转录活化子1(signal transducers and activators of transcription 1,STAT1)对Aβ_(1-42)诱导人神经母细胞瘤细胞系SH-SY5Y细胞损伤的影响。方法常规培养SH-SY5Y细胞系,应用Trypan染色进行形态学观察,Cell Counting Kit-8测定增殖活性;乳酸脱氢酶(lactate dehydrogenase,LDH)法检测毒力;荧光探针法测定氧活性;实时荧光定量多聚核苷酸链式反应方法检测miR-17-5p、STAT1 mRNA的表达;免疫印迹试验法检验STAT1基因的表达。结果与对照组比较,Aβ_(1-42)组细胞STAT1 mRNA与蛋白水平、增殖抑制率、细胞毒性、活性氧(reactive oxygen species,ROS)升高(P<0.05),miR-17-5p水平降低(P<0.05);与Aβ_(1-42)组比较,mimics NC组数据无统计学意义(P>0.05),miR-17-5p mimics组数据降低(P<0.05);与miR-17-5p mimics组比较,miR-17-5p mimics组+pcDNA3.1组数据无统计学意义(P>0.05),miR-17-5p mimics组+pcDNA3.1 STAT1组数据升高(P<0.05)。与对照组、mimics-NC组比较,miR-17-5p mimics组SH-SY5Y细胞中STAT1 mRNA与蛋白水平降低(P<0.05)。结论miR-17-5p的过表达可以作为STAT1的靶向负调节,从而对SH-SY5Y细胞的损害起到一定的保护作用。

Objective To investigate the effect of miR⁃17⁃5p overexpression on Aβ_(1-42)⁃induced injury in human neuroblastoma cell line SH⁃SY5Y by regulating signal transducers and activa⁃tors of transcription 1(STAT1).Methods SH⁃SY5Y cells were cultured.Trypan blue stai⁃ning was used for morphological observation,and Cell Counting Kit⁃8 was used to measure the proliferation activity.Lactate dehydrogenase(LDH)method was used to detect the toxicity.Fluorescence probe method was used to measure oxygen activity.Real⁃time fluorescent quanti⁃tative polynucleotide chain reaction was used to detect the expression of miR⁃17⁃5p and STAT1 mRNA.The expression of STAT1 gene was detected by Western blot.Results Compared with the control group,the levels of STAT1 mRNA and protein,proliferation inhibition rate,cyto⁃toxicity and reactive oxygen species(ROS)were increased(P<0.05),while the level of miR⁃17⁃5p was decreased(P<0.05)in the Aβ_(1-42) group.Compared with the Aβ_(1-42) group,the mimics NC group had no significant difference(P>0.05),and the miR⁃17⁃5p mimics group had a significant reduction(P<0.05).Compared with miR⁃17⁃5p mimics group,the data of miR⁃17⁃5p mimics group+pcDNA3.1 group had no significant difference(P>0.05),and the data of miR⁃17⁃5p mimics group+pcDNA3.1STAT1 group was increased(P<0.05).Com⁃pared with the control group and mimics⁃NC group,STAT1 mRNA and protein levels in SH⁃SY5Y cells in the miR⁃17⁃5p mimics group were decreased(P<0.05).Conclusion The o⁃ver⁃expression of miR⁃17⁃5p can act as a targeted negative regulator of STAT1 to protect SH⁃SY5Y cells from damage.