Effects of Fructus mume total flavone on miR-145-3p/CREB5 axis in SH-SY5 Y cells induced by MPP+
Objective To study the effects of Fructus mume total flavone(FMF)on gene expression of MPP+induced human neuroblastoma SH-SY5Y cells.Methods CCK-8 was used to screen the optimal concentrations and time points of FMF and MPP+acting on SH-SY5Y cells.SH-SY5Y cells from control group,model group(MPP+induction)and FMF group(MPP+induction+FMF intervention)were collected for high-throughput sequencing of whole gene transcriptome,and the differentially expressed miRNAs and mRNAs were screened.The potential target genes of miR-145-3p were analyzed by bioinformatics software,and intersected with differentially expressed mRNAs.SH-SY5Y cells were divided into 6 groups:control,model,miR-145-3p mimics,mimics NC,miR-145-3p inhibitor,and inhibitor NC.qRT-PCR was used to detect the expression level of miR-145-3p in cells.Western blot was used to detect the expression level of CREB5 protein in cells.And double luciferase reporter gene assay was used to detect the fluorescence activity of cells.Results There were 33 differentially expressed miRNAs between the control group and the model group,and 16 differentially expressed miRNAs between the model group and the FMF group.After taking the intersection,seven differentially expressed miRNAs were obtained,of which miR-145-3p was down-regulated in the model group and up-regulated in the FMF group.After analysis,four differential target genes regulated by miR-145-3p were obtained:CREB5,EGLN1,NTNG1,and SLC22A15.Compared with the control group,the expression level of miR-145-3p in the model group decreased significantly,and the expression level of CREB5 protein increased significantly.Compared with NC group,miR-145-3p expression level in miR-145-3p mimics group was significantly higher,CREB5 protein expression level was significantly lower,miR-145-3p expression level in miR-145-3p inhibitor group was significantly lower,CREB5 protein expression level was significantly higher.The results of double luciferase reporter gene showed that there was a targeted regulatory relationship between miR-145-3p and CREB5.Conclusion Both miR-145-3p and CREB5 are differentially expressed in SH-SY5Y cells induced by MPP+.FMF can regulate the expression level of miR-145-3p/CREB5 axis in cells.
Parkinsons diseaseFructus mume total flavonetranscriptome sequencingmiR-145-3pCREB5
万方数据
维普
