The Study Of Relationship Between SNPs In Genes Associated With VLDL/LDL Metabolic Pathway And Cerebral Hemorrhage In Han Population Of Changsha Area

Background and objective:Cerebrovascular disease is among the three leading causes of death worldwide,of which cerebral hemorrhage(CH) represents 10%to 55% with high morbidity and disability.CH is a heterogeneous disease caused by different pathogenic mechanisms.There are probably many alleles with small effect sizes interacting with environmental factors. Identification of gene polymorphisms that confer susceptibility to CH and the development of genetic risk diagnosis systems may contribute to the personalized prevention of these conditions.Atherosclerosis is an important risk factor of CH.It is acknowledged that very low density lipoprotein(VLDL) and Low density lipoprotein(LDL) are important risk factors for atherosclerosis. Imbalance disturbance of endogenous VLDL/LDL metabolic pathway may influence VLDL/LDL levels and the processe of atherosclerosis, which in turn may increase the risk of CH.The relationship between genes associated with endogenous VLDL/LDL metabolic pathway and CH has been rarely reported.The aim is to study the relationship between genes associated with endogenous VLDL/LDL metabolic pathway and CH in Han people of Changsha area,which include apolipoprotein AⅠ-CⅢ-AⅣ(ApoAⅠ-CⅢ-AⅣ),apolipoprotein B(ApoB),lipoprotein lipase(LPL),low density lipoprotein receptor(LDLR),and proprotein convertase subtilisin/kexin type 9(PCSK9) genes.Methods:All subjects came from Changsha Han population which were composed of 3 groups:(1) 200 members of 38 family with CH aggregation,including:①61 CH patients with family history:34 men and 27 women,averagely aged 53.26±10.60 years;②95 first-degree relatives:50 men and 45 women,averagely aged 51.15±14.45 years;③23 second-degree relatives:9 men and 14 women,averagely aged 36.82±18.68 years;④21 relatives with no kinship with proband:12 men and 9 women,averagely aged 52.32±12.74 years,who are spouses of the proband or the first-degree and second-degree relatives.(2)Sporadic cerebral hemorrhage(SCH) group:273 patients with sporadic cerebral hemorrhage,138 men and 135 women,averagely aged 55.38±12.08 years.(3) Control group:140 healthy controls,74 men and 66 women, averagely aged 53.62±13.28 years.Serum lipid levels were tested:Total cholesterol(TC),High density lipoprotein(HDL) and triglycerides(TG) were mesured by standard enzymatic methods.LDL cholesterol levels were calculated with the Friedewald formula(LDL=TC-HDL-TG/5).Multiplex SNaPshot genotyping analysis and DNA sequencing were used to detect 12 SNPs in 5 genes associated with endogenous VLDL/LDL metabolism of all subjects,including:rs12721026,rs2854116,rs45487004,rs670 of Apo AⅠ-CⅢ-AⅣgene,rs1367117,rs11279109 of ApoB gene,rs320 of LPL gene,rs2569542,rs688,rs5925 of LDLR gene and rs11206510,rs505151 of PCSK9 gene.The frequencies of the alleles and genotypes between the SCH group and control group were analyzed by x~2 analysis,and lipid levels were compared among genotypes by ANOVA using SPSS.The SNP and haplotype transmissions of the CH pedigrees were analyzed by transmission disequilibrium test(TDT) using FBAT computer program.Results:1.①There are apoAⅠ-CⅢ-AⅣgene rs12721026,rs2854116, rs45487004 and rs670 polymorphisms in Changsha Han population.②There is no significant difference of genotype and alleles frequencies of rs12721026,rs2854116,rs45487004 and rs670 between SCH group and control group(P＞0.05).③Application of TDT to genotype data from rs12721026,rs2854116,rs45487004 and rs670 polymorphisms revealed no evidence for transmission disequilibrium with CH(P＞0.05).④Haplotype analysis of rs12721026,rs2854116,rs45487004 and rs670 revealed no evidence for transmission disequilibrium with CH(P＞0.05).⑤Both in SCH group and in control group,the levels of HDL of the TT gene type of rs12721026 were significantly higer than that of the GT/GG gene type(p＜0.05),while the levels of HDL of the AA gene type of rs670 were significantly higer than those of the GG,GA gene type.2.①There are ApoB gene rs1367117 and rs11279109 polymorphisms in Changsha Han population.②There is no significant difference of genotype and alleles frequencies of rs1367117 and rs11279109 between SCH group and control group(P＞0.05).③Application of TDT to genotype data from rs1367117 and rs11279109 polymorphisms revealed no evidence for transmission disequilibrium with CH(P＞0.05).④Haplotype analysis of rs1367117 and rs11279109 revealed no evidence for transmission disequilibrium with CH(P＞0.05).⑤Both in SCH group and in control group,the levels of TC of the DD/DI gene type of rs11279109 were significantly higer than those of theⅡgene type,while the levels of TC of the GA/AA gene type of rs1367117 were significantly higer than that of the GG gene type(p＜0.05).3.①There are LPL gene rs320 polymorphism in Changsha Han population.②There is significant difference of genotype of rs320 between SCH group and control group(x~2=6.95,p=0.031).The frequencies of T allele were significantly higher in SCH group than these in control group(x~2=5.05,p=0.025).③TDT analysis revealed that the frequency of transmission(from parent to the offspring with CH) of alleles C of rs320 polymorphism in CH increased significantly(Z=2.144, p=0.034).④The levels of TG of the TT/GT gene type of rs320 in SCH group was significantly higer than that of the GG gene type(p＜0.05).4.①There are LDLR gene rs2569542,rs688 and rs5925 polymorphisms in Changsha Han population.②There is no significant difference of genotype and alleles frequencies of rs2569542,rs688 and rs5925 polymorphisms between SCH group and control group(P＞0.05).③Application of TDT to genotype data from rs2569542,rs688 and rs5925 polymorphisms revealed no evidence for transmission disequilibrium with CH(P＞0.05).④Haplotype analysis of rs2569542, rs688 and rs5925 revealed no evidence for transmission disequilibrium with CH(P＞0.05).⑤There was no significant difference of levels of TG,TC,HDL and LDL of different gene types of rs2569542,rs688,rs5925 in SCH group and control group(p＜0.05).5.①There are PCSK9 gene rs11206510 and rs505151 polymorphisms in Changsha Han population.②There is no significant difference of genotype and alleles frequencies of rs505151 between SCH group and control group(P＞0.05).The frequencies of CC/CT genotype and C allele of rs11206510 were significantly higher in SCH group than these in control group(x~2=4.19,p=0.041;x~2=4.28,p=0.039).③The levels of LDL of the GA/GG gene type of rs505151 were significantly higer than those of the AA gene type both in SCH group and in control group(p＜0.05).Conclusions:1.A allele in LPL gene rs11206510 polymorphism and C allele in PCSK9 gene rs11206510 polymorphism are associated with risks of CH in Changsha Han population. 2.There is no association of ApoAⅠ-CⅢ-AⅣgene rs12721026,rs2854116,rs45487004,rs670,apoB gene rs1367117,rs11279109, LDLR gene rs2569542,rs688,rs5925 and PCSK9 gene rs11206510 polymorphisms with CH in Changsha Han population.3.There is no association of haplotypes of ApoAⅠ-CⅢ-AⅣgene rs12721026,rs2854116,rs45487004,rs670,ApoB gene rs1367117,rs11279109,LDLR gene rs2569542,rs688,rs5925 polymorphisms with CH in Changsha Han population.4.ApoAⅠ-CⅢ-AⅣgene rs670,rs12721026,ApoB gene rs11279109,rs1367117,LPL gene rs320,PCSK9 gene rs505151 polymorphisms may be associated with lipid metabolism in Changsha Han population.