| Objective: Parkinson's disease(Parkinson's disease,PD)is a common neurodegenerative disease characterized by symptoms such as resting tremor,bradykinesia,stiffness,and postural instability.Recent studies have shown that genetic polymorphisms are closely related to the risk of sporadic Parkinson's disease,but the results of studies in different regions and races are still controversial.This study mainly explored LRRK2,APOE,FGF20,SNCA,PARK16,PITX3,BST1,PARKIN S/N,GBK and GBA gene polymorphisms are associated with sporadic Parkinson's disease in Jiangsu,China.And the Meta-analysis method was used to further explore the relationship between gene polymorphisms and the correlation and clinical symptoms in Chinese patients with Parkinson's disease.In addition,this study used matrix-assisted laser analytical ionization time-of-flight mass spectrometry,immunofluorescence PCR,and DNA sequencing to genotype some loci in order to compare the reliability and accuracy of the methods,at the same time,a more effective and quick method is selected for clinical application to assist the clinical diagnosis of Parkinson's disease.Methods: 1.Select 171 patients with sporadic parkinson's disease and 94 healthy controls in Jiangsu area as the research objects.The PD group was divided into earlyonset PD group and late-onset PD group according to the age of onset.Divided into male PD group and female PD group according to gender.2.Matrix-assisted laser desorption / ionization-time of fligh(MALDI-TOF MS)was applied to LRRK2(rs34778348,rs2046932,rs33949390),APOE(rs429358 and rs7412),FGF20(rs591323 and rs1721100),SNCA(rs356219,rs356220,rs894278,rs2736990,rs356182),PARK16(rs947211,rs823156,rs823128),PITX3(rs3758549),BST1(rs4698421),PARKIN S/N(rs1801474),GBK(rs1564282)and GBA(rs421016)genotyping common SNPs of genes,and three SNPs were randomly selected and analyzed by MALDI-TOF MS,fluorescent immuno-PCR,and sequencing.The accuracy of the three methods was compared.3.Use spss statistical software to processand analyze the genetic testing related data to explore the association between gene polymorphisms and the risk of PD.4.The Meta-analysis was used to further explore the association between LRRK2 ? GBA gene polymorphisms and sporadic Parkinson's disease,and related clinical symptoms in Chinese population.Results: 1.Compared with the healthy control group,the LRRK2 gene rs34778348 locus PD group had statistically significant differences in allele frequency and genotype frequency(OR = 5.71,P = 0.001;OR = 5.94,P = 0.010).Early-onset PD group,lateonset PD group and female PD group A allele frequencies were significantly higher than the control group,the difference was statistically significant.The age of onset of PD with genotypes AG /AA was about 5 years earlier than that of GG PD(P = 0.011),and female PD populations were more likely to mutate(P = 0.019),but there were no significant differences in male PD group,clinical symptoms and onset of disease.2.LRRK2 gene rs2046932 locus PD group compared with the control group,allele and genotype frequency distribution were statistically different(OR = 4.12,P = 0.005;OR = 4.23,P = 0.005).The frequency of A allele was significantly higher in the earlyonset PD group,late-onset PD group and female PD group than in the control group,the difference is statistically significant.The age of onset of PD in genotypes AG and AA was about 6 years earlier than that of GG-type PD(P = 0.002),and women with PD were more likely to mutate.However,no significant difference was found in male PD group,course of disease,and symptoms of onset(P> 0.05).3.Compared with the control group in the case group of GBA gene rs421016,the allele frequency and genotype frequency were statistically significant(P = 0.010;P =0.03).The A allele of the early-onset PD group was statistically different from the control group(P = 0.018),but there was no statistical difference in the analysis of gender,late-onset PD group,course of disease,and onset of symptoms(P > 0.05).4.The genotype frequency of the PARK16 gene(rs823128 locus)in the male PD group was significantly different from the control group(AG + GG / AA,OR = 0.44,95%CI 0.21-0.90,P = 0.024).However,no statistical analysis was found in other subgroup analyses.5.LRRK2 gene(rs33949390),APOE gene(rs591323),SNCA gene(rs1721100,rs356219,rs356220,rs894278,rs2736990,rs356182),PARK16(rs947211 and rs823156),PITX3(rs3758549),BST1(rs4698421),PARKIN S/N(rs1801474),and GBK(rs1564282)compared with the control group,the genotype and allele frequencieswere not statistically different(P > 0.05),and no statistical difference was found in further gender and age subgroup analysis(P > 0.05).6.MALDI-TOF MS,fluorescent PCR,and sequencing methods were used to detect SNP sites.The results of PCR and mass spectrometry were completely the same as those of gene sequencing,and the accuracy of the three methods was consistent.MALDI-TOF MS was more economical and high-throughput..7.The Meta-analysis showed that 44 studies were included,including 27,155parkinson's patients and 31,599 healthy controls.(1)A Meta-analysis of LRRK2 gene mutations revealed that LRRK2 gene polymorphisms were significantly associated with PD susceptibility in the Chinese population(OR = 2.02,95%CL,1.76-2.32,P = 0.000).LRRK2 gene SNP locus R1628P(C VS G: OR = 1.86,95%CL,1.63-2.11,P = 0.000),G2385R(A VS G: OR = 2.28,95%CL,1.86-2.79,P = 0.000),A419V(T VS C: OR =2.00,95%CL,1.02-3.93,P = 0.044)polymorphism was significantly associated with an increase in PD risk in the Chinese population,and there was no significant correlation between P755 L polymorphism and PD risk(P = 0.704).In terms of clinical symptoms,H-Y scores of PD patients with LRRK2 mutations are slightly lower than those of noncarriers,and MMSE scores are slightly higher than those of non-carriers(H-Y: MD =-0.06,95%CI,-0.11 ?-0.02,P = 0.009;MMSE : MD = 0.40,95%CI,0.17-0.63,P =0.001);UPDRS-?,UPDRS-?,and UPDRS-? scores were lower in the carrying group than in the non-carrying group(UPDRS-?: MD =-0.21,95%CI,-0.35 ?-0.06,P = 0.005;UPDRS-?: MD =-0.61,95%CI,-1.08 ?-0.14,P = 0.011;UPDRS-?: MD=-1.06,95%CI,-1.80 ?-0.32,P = 0.005),the difference is statistically significant.There was no statistically significant difference in the onset symptoms.(2)A Metaanalysis of GBA gene mutations revealed that GBA gene polymorphisms were significantly associated with PD susceptibility in the Chinese population(OR = 6.41,95%CL,3.60-11.40,P = 0.000),GBA gene SNP locus L444P(T VS C: OR = 7.17,95%CL,3.8-13.30,P = 0.000)polymorphism was significantly associated with an increase in PD risk in the Chinese population,and there was no significant statistical difference between the N370 S and R120 W polymorphisms in PD patients and the control group.In terms of clinical symptoms,Parkinson's H-Y score and UPDRS score were not significantly different between the GBA gene mutation-bearing group and the non-carrying group.Conclusions: 1.The rs34778348 locus of the LRRK2 gene is one of the risk factors for sporadic PD,and its allele A increases the risk of PD.The age of onset of patients with A allele is about 5 years earlier than other PD patients.This difference is more pronounced in female PD populations.Female PD populations with A alleles have an onset of about 5.53 years before other female PD patients.2.The LRRK2 gene rs2046932 locus is one of the risk factors for sporadic PD.Its allele A increases the risk of PD.Patients carrying the A allele are about 6 years earlier than other PD patients.Female PD populations are more likely to mutate.Female PD populations carrying the A allele It is about 6.19 years earlier than other PD patients.This study found that rs2046932 was related to the age and gender of Parkinson's disease.3.Allele G at the rs421016 locus of GBA gene can increase the risk of PD,and this locus is one of the risk factors for PD.The allele G at the rs823128 locus of the PARK16 gene may be a protective gene for Parkinson's disease.4.LRRK2 gene(rs33949390),APOE gene(rs429358 and rs7412 loci),FGF20(rs591323 and rs1721100),SNCA gene(rs356219,rs356220,rs894278,rs2736990 and rs356182),PARK16 gene(rs947211,rs823156),PITX3 gene(rs3758549),BST1 gene(rs4698421),GBK gene(rs1564282),parkin s/n167(rs1801474)have nothing to do with the onset of PD.5.The results of PCR and mass spectrometry detection of SNP loci were completely consistent with the results of gene sequencing.6.LRRK2 and GBA gene polymorphisms significantly increase the risk of sporadic Parkinson's disease in the Chinese population.PD patients with LRRK2 mutations have lower H-Y scores,UPDRS scores,and higher MMSE scores than non-carried patients.There were no significant differences in initial symptoms;PD patients with GBA mutations had no correlation in H-Y scores and UPDRS scores compared with non-carriers. |
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