The Association Between The Acute Large Artery Atherosclerosis Ischemic Stroke And The Polymorphism Of Aldehyde Dehydrogenase 2

BackgroundLarge Artery Atherosclerosis(LAA)cerebral infarction is the main clinical subtypes of ischemic stroke,refers to a cerebrovascular accident that occurs due to an interruption of blood supply to the brain which leads to dysfunction or death of brain cells.Today,stroke is a major cause of death and disability worldwide,so it is important to discuss the mechanism and treatment of LAA ischemic stroke.Mitochondrial aldehyde dehydrogenase 2(ALDH2)is a key enzyme that metabolizes acetaldehyde(a toxic intermediate of ethanol metabolism)to acetic acid.The ALDH2*2 variant has been implicated in various diseases,including alcohol related diseases such as hepatitis,hepatocirrhosis,digestive system tumors,alcoholic cardiomyopathy,cancer and coronary artery disease.ALDH2 is related to nervous system disease such as Parkinson's disease,and Alzheimer's disease and nervous system degeneration disease.However the study about cerebral infarction were few.Therefore,the research on the correlation between ALDH2 gene polymorphism and cerebral infarction and cerebrovascular disease,which is of great significance for early genetic diagnosis and targeted prevention and treatment.Objectives1.To investigate the relationship between the Glu504Lys polymorphism and the risk of acute cerebral infarction of Han Chinese in Shandong Province;2.To summarize the clinical features of LAA infarction,cerebrovascular lesions features,and collateral circulation,and then to reveal the correlation between the Glu504Lys genepolymorphism and them;3.To evaluate the relation of ALDH2 gene polymorphism and short-term prognosis in patients with LAA cerebral infarction by follow-up visits.Methods1.SubjectsCase-control study enrolled the acute ischemic stroke patients from January 2014 to September 2015 in Yantai Yuhuangding hospital as the case group,who were confirmed by clinical information and brain magnetic resonance imaging(MRI),TOAST classification of LAA,non-heavy drinking,complete DSA.Control group for the same period and area population doing health physical examination,excluding heavy drinkers.2.Clinical dataBasic clinical information,including age,sex,family history of hypertension,diabetes,cerebrovascular disease history,were recorded.Venous blood samples were taken from each subject.The levels of total cholesterol,triglyceride,uric acid,and homocysteine were analyzed using routine blood and biochemistry tests.All case-patients in the hospital the same day by two trained specialists for NIHSS score on neurologic impairment.3.Cerebrovascular evaluationTwo experienced neuroradiologists,who were blinded to all clinical information,reviewed digital subtraction angiography for each case group participant to assess complete vessel conditions and collateral circulation.We collected the data of lesion location and collateral circulation.4.ALDH2 genotypingGenomic DNA was extracted from venous blood samples using a commercial DNA extraction kit(TIANGEN,affiliated to QIAGEN,Beijing,China).PCR amplification was performed using ALDH2 primers in a 50 pL PCR mixture containing 2 ?L DNA template,10?L primers,and 1 U Tag DNA polymerase for an initial denature step of 94? for 3 min and then 35 cycles of 94? for 45 s,60? for 30 s and 72? for 45 s and final extension of 72? for 5 min.The PCR products were then purified using a commercial kit(TIANGEN)and were directly sequenced by Invitrogen Corporation(Nanjing,China).5.PrognosisAcute cerebral infarction patients were evaluated daily living conditions on 90 days follow-up visits,assessment scales used modified Rankin scale(mRS),by two trained professionals neurologists.6.Statistical analysisAll statistical analyses were performed using the SPSS 23.0 statistical package(SPSS Inc.,Chicago,IL,USA).An x2-test was used to determine whether the ALDH2 genotypes were in Hardy-Weinberg equilibrium.Differences between groups for continuous values were calculated using Student's t-test.For dichotomous variables,the chi-square(?2)test and logistic regression analysis were applied.The criterion for statistical significance was a p-value<0.05.Results1.Clinical dataThe number of patients exhibiting a family history of cerebrovascular disease(P=0.033),diabetes mellitus(P =0.034),homocysteine(P =0.044),and total cholesterol(P =0.033)were compared to the controls.2.Cerebrovascular evaluation366 of cerebral infarction patients with vascular stenosis rate than 50%by DSA examination,which had stenosis of intracranial-extracranial artery 112 cases(28.43%),simple extracranial artery stenosis in 78 cases(19.8%),simple intracranial arterial stenosis in 176 cases(44.67%).366 cases of DSA has positive results were found in patients with intracranial atherosclerotic stenosis 691.In all lesions vascular,there were 394 stenosis in anterior circulation(57.02%)and 297 in posterior circulation(42.98%).There were most stenosis in extracranial segment of VA(24.31%),second for MCA(19.68%).178 cases opened collateral circulation(46.2%),most of them were with occlusion of blood vessels(135,75.84%).As the stenosis degree were worse,opened collateral circulation levels were more.3.ALDH2 gene polymorphismThe data generated showed that the frequency of the ALDH2 genotype did not differ between patients and controls(P =0.71).We performed subgroup analyses for the DSA abnormalities(EAS subtype,IAS subtype,IC-EC subtype).After taking into account adjustments for the patients'conventional risk factors,there was a significantly lower frequency of the ALDH2 AA genotype in patients with IAS abnormalities(OR 0.18,95%CI 0.04-0.81,p = 0.03).However the frequency of AA genotype was significantly higher in patients with abnormal EAS(OR 6.05,95%CI 1.16-31.53,p = 0.03).The rate of good collateral vessels was significantly lower in the mutation group(GA+AA)(OR 3.00,95%CI 1.54-5.82,p = 0.001).The allele frequency was also significantly different between two subgroups(OR 1.86,95%Cl 1.09-3.16,p = 0.02).The two groups were with a statistically significant difference on the opening of the collateral(?2=6.542,P=0.038).4.Prognosis314 cases finished 90 days follow-up visits.ALDH2 Glu504Lys genotype has a significant correlation with 90-day prognosis of patients(OR 0.530,95%Cl 0.333-0.841,P=0.007).This correlation was weakened after adjustment for the history of diabetes(P=0.031),poor opened collateral circulation(P=0.001),NIHSS score?6(P=0.000).However,the association between ALDH2 genotype and short-term prognosis was still significant(OR 0.570,95%CI 0.326-0.998,P=0.049).Conclusions1.The number of patients exhibiting a family history of cerebrovascular disease,diabetes mellitus,homocysteine and total cholesterol were compared to the controls of Han Chineses in Shandong Province.2.The number of intracranial vascular stenosis was more than extracranial vascular stenosis in acute ischemic stroke patients of Han Chineses in Shandong Province.3.The number of anterior circulation stenosis was more than posterior circulation in acute ischemic stroke patients of Han Chineses in Shandong Province.Appear in narrow frequency order for EC-VA>MCA>EC-ICA>IC-VA>IC-ICA>ACA>BA>PCA>CCA?4.The first collateral circulation was most in collateral good case;as the stenosis degree were worse,opened collateral circulation levels were increased.5.The frequency of the ALDH2 genotype did not differ between patients and controls in acute ischemic stroke patients of Han Chineses in Shandong Province.6.ALDH2*2/*2 genotype is an independent protective factors for intracranial arterial stenosis and ALDH2*2/*2 genotypes are independent risk factors for extracranial artery stenosis in acute ischemic stroke patients of Han Chineses in Shandong Province.7.The rate of good collateral vessels was significantly lower in the mutation group in acute ischemic stroke patients of Han Chineses in Shandong Province;ALDH2 activity effected the collateral circulation level.8.The relative risk of poor 90-day prognosis in patients with ALDH2 mutation was higher than wild group.9.The correlation of ALDH2 Gu504Lys genotype and 90-day prognosis was weakened but still significant after adjustment for the history of diabetes,poor opened collateral circulation and NIHSS score>6.