| ObjectiveIntracranial aneurysm(IA) is abnormal local distension of intracal artery, occurring as the consequence of the local discrepancy of the vessel wall and higher pressure in lumens.Incidence of ruptured IA is 18—23/100000/year that can result in subarachnoid hemorrhage(SAH),Ruptured IA is the third in the patients of cerebral vascular accident,after cerebral thrombosis and brain hemorrhage resulted from hypertension,especially from 40 to 60 years old.IA is more common in female more than in male,and female post-menopause is more than that of pro-menopause.Smoking,drinking, hypertension,hyperlipemia,hyperglycosemia are the risk factors of IA.SAH resulted from IA is a disease which can result in higher mutilation and higher mortality,12%of patients die before therapied,40%of hospitalized patients die in a month after haemorrhage,more than 1/3 patients remain severe nerval functional disturbance,only few patients recoveries well.The incidence of non-disruptured IA is 2.7—6.5%according to the autopsied and angiographic evaluation.Epidemiological investigation discovers that parts of patients who are with IA present familial propensity and concurrence immune disease,hereditary connective tissue disease and systemic disease.That elucidates genetic factor plays an important role in the development of IA.In pathogenesis,IA is considered a multigenic disease which is simultaneouly effected by genetic factors and environmental factors, complicately and uncontrollably.It consistents with either autosomal dominant inheritance,autosomal recessive heredity or X linkage heredity,assignment of genes is more disparity,so the etiopathogenesis of IA is not clear,that increases the difficulty in etilogical treatment.At present,the study of multigenic disease in genetics are linkage analysis and correlation study,the latter is divided into allelic association and linkage disequilibrium analysis.It promotes the analysis of DNA mutations and the localization of predisposing genes and the study of clone in major of multigenic diseases accompanying human complete sequence of genome obtaining in successfully, international human haplotype map plan of genome enforced,a great quantity of tagged single nucleotide polymorphisms(SNPs)used,genie array and microsatellite developed and applicated,the characteristic of predisposing genes is that genie mutation can result in increasing ailing risk to organism and not the generation of disease,it can cause it once environmental adverse factors intervene.so it is very important in locating predisposing genes.A lot of domestic and foreign scholars have detected more than ten predisposing genes of IA including in collogen gene,tumour necrosis factor receptor gene, elastin gene et al adoptting in research methods and located them.But their research papers are discrepancy,even opposite among different races,different nations and different crowds,that concludes that the etiopathogenisis of IA is uncertain.Up to now,SNP is the most useful method in revealling the association about liability between multiple genie change and some diseases such as cardiac disorder,diabetes,diabetes, schizophrenia and so on.There is favourable and extensive outlook in development and its craeteristic are quick,high performance and exact in SNP.So,it has become the content and target of human genome project in looking for and researching SNP. Finding out the pleiomorphic site relative with common disease on DNA sequence is one of the most important ways in comprehending complicate reasons which cause human disease.It will be possible in finding out the pathogenesy of multigenic disease including IA utmostly following with the continual development in molecular genetics and the application of various research methods.That offers the foundation of molecular genetics for definiting predisposing genes of IA,screening susceptible population,early diagnosis and early treatment and there have very momentous significance in elevating detection rate of disease,patient's survival rate and survival quality endlessly.Methods1.Epidemiological study of sporadic intracranial ruptured aneurysmA ease-control study including 100 patients of sporadic intraeranial ruptured aneurysm and 116 non-intracranial aneurysm controls was performed to analyze the effect of risk factors on the liability of IA.chi square test and Logistic regression model were used for multifactor analysis.2.Association analysis between the single nucleotide polymorphism of COL1A2 gene,COL4A1 gene,TNFRSF13B gene and microsatellite DNA marker D7S2421 and sporadic intracranial ruptured aneurysmsGenomic DNA was isolated from venous blood from randomly selected 100 unrelated patients with sporadic intracranial ruptured aneurysm and 116 controls of non-intracranial aneurysm.The difference of frequencies of genotypes between the two groups of COL1A2 SNP rs42524 G＞C,COL4A1SNP rs3783107 C＞T, TNFRSF13B SNP rs11078355G＞A and microsatellite DNA marker D7S2421 SNP rs6465976of G＞A were genotyped by polymerase chain reaction- restriction fragment length polymorphism(PCR-RFLP) assay,then analyzed by SPSS 13.0 to investigate the association between the genes and sporadic intracranial ruptured aneurysm.3.Clinical significance of single nucleotide polymorphism of COL1A2 gene, COL4A1 gene,TNFRSF13B gene and microsatellite DNA marker D7S2421 in the patients of sporadic intracranial ruptured aneurysm in the northeast of China.Genomic DNA was isolated from venous blood from randomly selected 100 unrelated patients with sporadic intracranial ruptured aneurysm.Polymorphisms of COL1A2 SNP rs42524 G＞C,COL4A1SNP rs3783107 C＞T,TNFRSF13B SNP rs11078355G＞A and microsatellite DNA marker D7S2421 SNP rs6465976of G＞A were genotyped by polymerase chain reaction- restriction fragment length polymorphism(PCR-RFLP) assay.The frequencies of genotypes were analyzed by SPSS13.0.All the data were inspected by chi square test(x~2 test),calculating P value,odds ratio(OR) and 95%confidence interval(95%CI) to analyze the association between the clinical features of sporadic intracranial ruptured aneurysms and the SNPs.Results1.Epidemiological study of sporadic intracranial ruptured aneurysmIn the100 IA patients,63.00%were at age of 40 to 59 years;monofactorial analysis showed that there was obvious difference between genders(P=0.036,OR= 1.794,95%CI 1.036-3.107),it was more common in female more than in male that were resulted in IA and there was statistical significance in smoking between the two groups(P=0.005,OR=2.327,95%CI 1.280～4.231);The rise of fasting blood glucose (FBG) and hypertension were associated with IA(P＜0.001,OR=4.114,95%CI 2.310-7.329;P=0.005,OR=2.161,95%CI 1.252-3.729,respectively).Multifactor weighting analysis showed that there was still obvious difference between genders(P＜0.001,OR=9.435,95%CI 3.403-26.163) and there was also statistical significance in smoking between the two groups(P＜0.001,OR=0.098,95%CI 0.035～0.275);The rise of FBG and hypertension were still associated with IA(P＜0.001,OR =4.019,95%CI 2.125-7.877;P=0.016,OR=2.195,95%CI 1.158-4.159,respectively).2.Association analysis between the single nucleotide polymorphism of COL1A2 gene,COL4A1 gene,TNFRSF13B gene and microsatellite DNA marker D7S2421 and sporadic intracranial ruptured aneurysmThe distribution of the polymorphisms of COL1A2 SNP rs42524 G＞C, COL4A1SNP rs3783107 C＞T,TNFRSF13B SNP rs11078355G＞A,microsatellite DNA marker D7S2421 SNP rs6465976of G＞A in cases(the group of sporadic intracranial ruptured aneurysm)and controls(the group of non- intracranial aneurysm) were consistent with a Hardy-Weinberg equilibrium,and the frequency of COL1A2 SNP rs42524 G＞C and TNFRSF13B SNP rs11078355G＞A genotypes were statistically significant between cases and controls(x~2=9.727,P=0.002,OR=2.479,95%CI 1.392-4.414;x~2=4.506,P=0.034,OR=1.870,95%CI 1.046-3.345).3.Clinical significance of single nucleotide polymorphism of COL1A2 gene, COL4A1 gene,TNFRSF13B gene and microsatellite DNA marker D7S2421 in the patients of sporadic intracranial ruptured aneurysm in the northeast of China.The study estimated the association between COL1A2 SNP rs42524 G＞C, COL4A1SNP rs3783107 C＞T,TNFRSF13B SNP rs11078355G＞A,microsatellite DNA marker D7S2421 SNP rs6465976of G＞A and the clinical features of sporadic intracranial ruptured aneurysm.(1) Among the patients of sporadic intracranial ruptured aneurysm,there were no associated between gender,age,blood pressure,blood glucose,Fisher's grade and the genotype of COL1A2 SNP rs42524 G＞C,while the frequency of COL1A2 SNP rs42524 GC+CC genotype was higher than that of the GG genotype among IA patients whose scale of Hunt and Hess(H&H)grading system were more than three(P=0.001, OR=9.265,95%CI 2.546-33.708).(2) There were no associated between gender,age,blood glucose,Hunt and Hess (H&H)grading system and the genotype of COL4A1 SNP rs3783107 C＞T,while the frequency of COL4A1 SNP rs3783107 CT+TTgenotype was higher than that of the GG genotype among IA patients whose blood pressures were more than 140/90 mmHg (P=0.035,OR=2.538,95%CI 1.056-6.100);in addition,the frequency of COL4A1 SNP rs3783107 CT+TT genotype was higher than that of the CC genotype among IA patients whose Fisher's grades were above two(P=0.009,OR= 5.000,95%CI 1.378-18.148).(3) There were no associated between gender,blood pressure,blood glucose, Hunt and Hess(H&H)grading system,Fisher's grade and the genotype of TNFRSF13B SNP rs11078355G＞A,while the frequency of the TNFRSF13B SNP rs11078355 GA+AA genotype was higher than the GG genotype among IA patients aged≥65 years(P=0.004,OR=9.771,95%CI 1.897-50.329).(4) There were no associated between gender,age,blood glucose,Fisher's grade and the genotype of microsatellite DNA marker D7S2421 SNP rs6465976 G＞A,while the frequency of microsatellite DNA marker D7S2421 SNP rs6465976 GA+AA genotype was higher than that of the GG genotype among IA patients whose blood pressures were more than 140/90 mmHg(P=0.026,OR=2.513,95%CI 1.105-5.712); in addition,the frequency of microsatellite DNA marker D7S2421 SNP rs6465976 GA+AA genotype was higher than that of the GG genotype among IA patients whose scale of Hunt and Hess(H&H)grading system were more than three(P=0.008,OR= 4.333,95%CI 1.401-13.401).Conclusions1.Sporadic intracranial ruptured aneurysm mostly occurs in people aged 40 to 59 years.Female,smoking,the rise of FBG and hypertension were the risk factors of IA.2.COL1A2 SNP rs42524 G＞C and TNFRSF13B SNP rs11078355G＞A are obviously associated with the sporadic intracranial ruptured aneurysm in the northeast of China,and COL1A2 gene and TNFRSF13B gene are the candidate predisposing genes.3.Among the patients of sporadic intracranial ruptured aneurysm:(1) The allele C of COL1A2 SNP rs42524 G＞C maybe raise the scale of Hunt and Hess(H&H)grading system and maybe the important risk factor that can aggravate the IA patients'pathogenetic condition;(2) The allele T of COL4A1 SNP rs3783107 C＞T maybe the risk factor of IA in the patients that are hypertension,at the same time,it maybe raise the degree of the patients' Fisher's grade;(3) The allele A of TNFRSF13B SNP rs11078355G＞A maybe the independent risk factor of IA excluding the senile diseases such as hypertension and diabete in old people;(4) The allele A of microsatellite DNA marker D7S2421 SNP rs6465976 G＞A maybe the risk factor of IA in the patients that are hypertension,and it maybe raise the scale of Hunt and Hess(H&H)grading system,aggravate the IA patients'pathogenetic condition. |
PDF Full Text Request