MiR-31 Participates In The Study Of The Pathogenesis Of Parkinson's Disease By Regulating The Expression Of SRSF11 Protein

PART? RNA Sequencing Analysis of mi R-31 in SH-SYSY CellsBackground & Objective: Parkinson's disease(PD)is an aggravated dyskinetic disorder.At present,the cause of PD is not clear and cannot be fundamentally cured.In this study,seven differentially expressed mi RNAs were screened by Solexa sequencing and q RT-PCR in the serum of patients and controls,and were further verified in RNA extracts from brain tissue.Finally,mi R-31 were identified.This part of the study continues the above studies to explore the role of mi R-31 in the signal pathway in human neuroblastoma cells(SH-SY5 Y cells),providing a theoretical basis for the following study of mi R-31 in PD.Methods: SH-SY5 Y cells in logarithmic growth phase were used,and mi R-31 mimic,mi R-31 inhibitor,mi R-NC mimic,and mi R-NC inhibitor were transfected into SHSY5 Y cells.Four groups of SH-SY5 Y cells were subjected to deep transcriptome sequencing by RNA-seq technology.R software was used to analyze differential gene expression in cells,GO function,Pathway function and transcription factor family of differential genes.Results: 1)The results showed that 841 genes were up-regulated and 1268 genes were down-regulated.2)GO analysis results showed that differential genes involved in cell processing,single organisms,metabolism,processes,biological regulation,composition of cellular components,protein binding and other processes.3)Pathway analysis shows that most of the differential genes are distributed in signal transduction,signaling molecules and interaction,tumor formation,viral infectious diseases,metabolism,immune system,endocrine system and other pathways.Of these,67 differential genes were involved in cell proliferation and death pathways.Conclusion: mi R-31 has important biological functions and is involved in signal transduction,cell proliferation and death,tumor formation,immunity,metabolism and other signaling pathways in cells,providing a preliminary basis for the study of mi R-31.Mi R-31 regulates its expression by binding to the 3? UTR region of SRSF11 and regulates the expression of apoptosis-related gene Bax/Bcl-2 m RNA to inhibit apoptosis.PART? Study on the function of mi R-31 in Parkinson's disease and its mechanismBackground & Objective: To investigate the mechanism of mi R-31 in the pathogenesis of PD by regulating the expression of SRSF11.Methods: First,human neuroblastoma cells(SH-SY5 Y cells)were treated with different concentrations of MPP + solution to construct an acute PD cell model;flow cytometry was used to investigate the effect of mi R-31 on MPP+-induced apoptosis of SH-SY5 Y cells;Prediction of mi R-31 target genes was identified by bioinformatics.Western blot and q RT-PCR were used to detect the intracellular expression of the target gene m RNA and protein.According to the results,the most likely target gene was selected.And then it was verified by the dual luciferase reporter assay whether mi R-31 directly binds to the 3?UTR region of SRSF11.Finally,a rescue experiment was performed to verify whether mi R-31 influences cell apoptosis by regulating SRSF11 m RNA expression.Results: 1)Flow cytometry and CCK8 results showed that mi R-31 had protective effects on MPP+-induced apoptosis of SH-SY5 Y cells.2)Bioinformatics predicts that SRSF11 is a target gene of mi R-31,and q RT-PCR and Western blot verify that mi R-31 can regulate the m RNA and protein expression of SRSF11.3)Double luciferase reporter assays confirmed that mi R-31 binds to the 3?UTR region of SRSF11 and thereby inhibits SRSF11 expression.mi R-31 is a direct target gene for SRSF11.4)Rescue experiments showed that mi R-31 and SRSF11 have opposite effects on apoptosis,which can regulate the expression of Bax/Bcl-2 m RNA and affect apoptosis.Mi R-31 can bind to the 3?UTR of SRSF11 to inhibit its expression and reverse the apoptosis caused by overexpression of SRSF11.Conclusion: Mi R-31 can resist the cytotoxicity of SH-SY5 Y cells caused by MPP+,reduce cell apoptosis and protect cells.Mi R-31 regulates the expression of SRSF11 by binding to its 3?UTR and inhibits m RNA and protein expression of SRSF11.SRSF11 is a direct target gene of mi R-31.Mi R-31 can down-regulate the expression of SRSF11 and regulate apoptosis-related gene Bax/Bcl-2 m RNA expression to inhibit apoptosis.PART? Study on the association between STK39?BST1 single nucleotide polymorphisms and the risk of Parkinson's disease and the progression of motor symptoms in Chinese populationBackground&Objective: While studying the genetic level of PD,we also studied the related risk genes in Chinese PD population.After consulting the relevant PD literature,we found that the single nucleotide polymorphisms(SNPs)of STK39 and BST1 genes are related to the onset risk of Caucasian PD.SNPs are affected by race and environment.Therefore,this part of the study was designed to investigate the association between the SNPs of the STK39 and BST1 genes and the risk of Parkinson's disease and the progression of motor symptoms in Chinese populationMethods: We collected 153 sporadic PD patients who came to the PD outpatient department of the Brain Hospital attached to Nanjing Medical University.At the same time,197 people from a healthy control group were collected from the Nanjing Physical Examination Center.The basic population data collection and evaluation of the Unified Parkinson's disease rating III(UPDRSIII)and the Hoehn-Yahr(H-Y)stage of the fifth part were performed by a professional neurological physician,and 5 m L of venous blood was collected.All subjects were typed for STK39 and BST1 genes using Mass ARRAY? SNP.Patients with PD were enrolled for follow-up(average duration of follow-up was greater than 2 years),and the PD scale was reassessed by a neurological physician.The mixed linear model and multiple linear regression model of the lme4 software package of R software were used to analyze the correlation between genotype,PD risk and PD disease progression.Results: 1)The single nucleotide polymorphism in the BST1 rs4698412 locus is associated with the pathogenesis of PD,and AA type is a risk factor for PD.The risk of developing PD of patients carrying the AA genotype was 1.54 times higher than that of the control group.2)Compared with the AG+GG type,the motor symptoms with AA type of STK39 rs2390669 locus was significantly faster.The genotypes of BST1 rs4698412 locus and rs1931532 locus were not significantly associated with the progression of motor symptoms.Both the rigidity score and the bradykinesia score of UPDRS III were significantly correlated with the genotype of the rs4698412 locus of the STK39 gene.The AA genotype at STK39 rs2390669 is a risk factor for accelerated progression of PD motor symptoms and can accelerate the progression of rigidity and bradykinesia symptoms.Conclusion: Single nucleotide polymorphisms in the BST1 rs4698412 locus are associated with the pathogenesis of PD.The single nucleotide polymorphism at STK39 rs2390669 is a risk factor for the faster progression of symptoms of rigidity and bradykinesia in PD patients.