Association Of HDAC9 Gene Polymorphic Loci With Susceptibility To Ischemic Stroke And TCM Syndrome Based On GWAS

ObjectiveThe protein deacetylase 9(HDAC9)gene which is screened out by Genome-wide association study(GWAS)in European population(3548 IS patients and 5972 controls)is associated with the susceptibility to ischemic stroke.There is a significant correlation between polymorphism and the attack of ischemic stroke in the study of people in the north of China.However,there is no same study on the population of the southern population,and there is no correlation between the polymorphism of the gene and the susceptibility of TCM syndrome.Therefore,we studied the association between the 2 loci of HDAC9 gene rs2240419 and rs2389995 polymorphism and ischemic stroke and its susceptibility to TCM syndromes in Guangdong Han population in order to study on the relationship between the single nucleotide polymorphisms of HDAC9 gene in Guangdong Han population and the susceptibility to ischemic stroke and TCM Syndrome.It also hopes to provide some ideas and foundations for further research on the influence of genetic mechanism on ischemic stroke,and provide some scientific explanation for the differentiation of TCM syndromes in stroke.MethodsA case control study is used in this study.We Select HDAC9 gene loci rs2240419 and rs2389995 as research sites.According to the inclusion criteria,184 cases in group IS and 116 cases in control group were included.Blood was extracted from all the subjects and then the genotyping was made after extracting DNA.At the same time,the trained investigators collected all the research object information,including general information,medical history,allergy history,personal history,family history,neurologic impairment score,TCM syndrome(IS only group of TCM syndrome type)and laboratory data,and through the research object of the self-designed observation table(table CRF)record.Finally,the results of the genotyping and the information of the CRF table were introduced into the Excel software to establish the database,and the statistical analysis was carried out in the SPSS 17 software.Results1.A total of 184 cases of IS were included in this study,including 112 males and 72 females,and 116 cases in the control group,including 55 men and 61 women.2.In TCM syndrome,141 cases of wind syndrome,13 cases of fire syndrome,121 cases of phlegm syndrome,149 cases of blood stasis and 82 cases of qi deficiency were obtained in group IS.3.Among all the subjects,rs2240419 locus(n=300)detected 3 genotypes of CC,CT and TT.Rs2389995 locus(n=291)detected 3 genotypes of AA,AG and GG.The three genotypes were represented by Nobu Juriko,heterozygote and mutant homozygote.4.To test for hardy Weinberg equilibrium in all subjects,the final results suggest that the IS group and the control group HDAC9 genotype distribution of SNP gene rs2240419 and rs2389995 were consistent with the Hardy-Weinberg equilibrium(P>0.05).It is suggested that the subjects of the control group used in this study are random population distribution,which is representative of the group of the same Mendel population.5.The comparison of the basic characteristics between the IS group and the control group:(1)The difference of gender composition was statistically significant(P<0.02),and there was no significant difference in age,age grouping and BMI composition(P>0.05).(2)Comparison of history and personal history of the group,the difference was statistically significant visible previous stroke,hypertension,diabetes,previous history of coronary heart disease,previous high HCY,drinking and smoking histories of the 7 project(P<0.05),and were positively correlated with ischemic stroke occurred;there was no significant difference with previous history of TIA dyslipidemia,allergy history of these 3 projects(P>0.05).(3)In the comparison of laboratory test results,there were statistically significant differences between the 6 items of SBP,DBP,HB,TG,HDL and HCY(P<0.05),and the difference between the other items was not statistically significant(P>0.05).(4)The results of multivariate analysis showed that age,HDL and SBP of the 3 groups was statistically significant difference between the age of onset(P<0.05),and HDL and stroke was negatively correlated with the incidence of stroke of SBP was positively related.The age and SBP of the 2 confidence intervals are small,so the degree of correlation may not significant.6.The differences in gene polymorphism between the IS group and the control group were compared.(1)The results of single factor and multi factor analysis showed that there was no significant difference between the genotypes(P>0.05).(2)By constructing 5 genetic models,we performed Logistic regression analysis for ischemic stroke patients(group IS)and control group.The results showed that there was no significant difference between the polymorphism of locus rs2240419 and rs2389995 locus and ischemic stroke(P>0.05).7.the basic characteristics of the non wind syndrome group(n=43)and the wind syndrome group(n=141)were compared.(1)There was no significant difference in gender,age,age grouping,BMI,previous history,personal history and laboratory test(P>0.05).(2)There was no statistical significance in the multi factor analysis(P>0.05).8.The results of the basic characteristics of the non phlegm syndrome group(n=63)and the phlegm syndrome group(n=121)were as follows:(1)There was no significant difference in gender,age,age grouping,BMI,previous history and individual history group(P>0.05).(2)Compared with the laboratory test group,the difference between the 3 items of HB,TG and TC was statistically significant(P<0.05),and the difference between the other 9 items was not statistically significant(P>0.05).(3)Multivariate analysis showed that there was a statistically significant difference between the 2 groups in TG and allergy history(P<0.05).TG was positively correlated with phlegm syndrome and had a certain degree of correlation,while the history of allergy was negatively correlated with phlegm.9.The basic characteristics of non blood stasis syndrome group(n=35)and blood stasis syndrome group(n=149)were compared.(1)There was no statistically significant difference in gender,age and age grouping,past history,personal history and laboratory test results(P>0.05).There was a statistically significant difference in BMI composition(P<0.05).(2)The results of multi factor analysis showed that there were statistical and positive correlation between the groups of BMI,but the range of the confidence interval was small and the degree of correlation may not be significant.10.The results of the differences in gene polymorphism among the groups were compared.(1)Univariate and multivariate analysis results indicated that there was no significant difference in the syndromes between each group(non wind syndrome group and wind syndrome group,non phlegm syndrome group and phlegm syndrome group,non blood stasis syndrome group and blood stasis syndrome group)genotype(P>0.05).(2)By constructing 5 genetic models of various syndromes of stroke group(non wind syndrome and wind syndrome,phlegm syndrome and phlegm syndrome,blood stasis syndrome and non blood stasis syndrome)are the research object of Logistic regression analysis results showed that the polymorphism of rs2240419 and rs2389995 of the HDAC9 gene in each syndrome group(non wind syndrome and wind syndrome group and non phlegm group and non phlegm syndrome group,blood stasis group and blood stasis group)had no significant difference(P>0.05).ConclusionFor the Han population in Guangdong:1.Male,past stroke,hypertension,diabetes,coronary heart disease,high HCY,drinking history and smoking history may be risk factors for ischemic stroke.2.In the laboratory test results,SBP,DBP,HB,TG,HDL and HCY may be the factors affecting the ischemic stroke.HDL may be a protective factor for ischemic stroke.3.The TCM Syndromes of ischemic stroke patients may be associated with HB,TG,TC and previous allergy history.The increase of TG may increase the possibility of the appearance of phlegm syndrome.4.The syndrome of traditional Chinese medicine(TCM)of ischemic stroke may be associated with the BMI index.5.The polymorphism of the rs2240419 and rs2389995 loci of the HDAC9 gene may not be related to the susceptibility to ischemic stroke.6.The polymorphism of SNP locus rs2240419 and rs2389995 in HDAC9 gene may not be related to the susceptibility of TCM Syndromes of stroke(wind syndrome,phlegm syndrome and blood stasis syndrome).7.The results of this study and the European region and northern region of similar China loci reported different results,and no positive findings,but can not be denied a clear such a susceptibility gene polymorphism and ischemic stroke and its TCM syndrome does not exist in association.It is not excluded that this research is different from the reference research in research subjects,interference factors control,etc.,or there are some limitations and shortcomings in this study,resulting in negative results.8.From the current research for syndrome gene level is still in the initial stage of development,the next step is necessary to solve these limitations and shortcomings of the perfect plan,for further research on correlation between susceptibility gene polymorphism and ischemic stroke syndromes and provide more high-quality and effective research program.