摘要
原发性高血压(EH)具有高度的遗传异质性、迟显性和易感基因外显不全性,发病机制复杂。越来越多的资料显示原发性高血压患者体内出现细胞免疫和体液免疫紊乱,HLA抗原分子可能参与了血压调节。多种血清学和基因型的关联分析及家系连锁分析均显示HLA-DRB1抗原特异性或等位基因与EH的易感性相关,但存在种族和地区特异性。近年来,单核苷酸多态性的检测在疾病易感基因定位中得以广泛应用,但目前国内外关于HLA-DRB1的SNPs与原发性高血压关系的报道还相对较少。
本研究通过对HLA-DRB1基因的rs2308765、rs9269186和rs3135388三个位点的单核苷酸多态性与原发性高血压关系的研究,进一步探讨HLA-DRB1基因的单核苷酸多态性与原发性高血压是否存在某种关联,从而为高血压发病的遗传学机制研究提供新的线索,也为高血压的临床治疗奠定理论依据;同时通过对原发性高血压内环境因素的单因素和多因素分析及环境与基因之间交互作用研究,综合探讨了原发性高血压的影响因素,为提出合理的预防控制措施提供参考依据。
结果显示:rs2308765位点的单核苷酸多态性与原发性高血压存在关联,G→T突变可能是原发性高血压的危险因素;rs9269186位点和rs3135388位点的单核苷酸多态性与原发性高血压无关;除低密度脂蛋白具有边缘统计学差异外,血糖、总胆固醇、甘油三酯及高密度脂蛋白在病例组和对照组间均具有统计学差异;病例组BMI、腰围、血糖、总胆固醇、甘油三酯及低密度脂蛋白均高于对照组,高密度脂蛋白低于对照组;高BMI和高甘油三酯是原发性高血压的危险因素;rs2308765位点的多态性与甘油三酯存在负相加交互作用。
Essential hypertension (EH) is a complex disease with multi-gene and multi-environmental trait, accounting for more than 90% in hypertension. Recently, the prevalence of hypertension showed a increasing trend all over the world. It has forecasted that, by 2025 there will be 1.58 billion hypertension patients in the world. According to the epidemiological survey in the 10th Five-Year Plan, the standardized prevalence of hypertension in China has reached 24.4%. There are more than 200 million patients with hypertension in China. It has estimated that approximately one million premature deaths had relationship with hypertension every year. The prevention and control of hypertension is a very formidable task in China. Therfore, while the important significance of prevention and treatment of hypertension are stressed, the etiologic research in hypertension is also a hot topics.
Hypertension has the characteristics of highly genetic heterogeneity, delayed dominant and complex pathogenesis. Increasing evidence shows that many patients with essential hypertension showed a cellular and humoral immune disorder, so the conclusion has been drawed that HLA antigen might play an important role in blood pressure regulation. Single nucleotide polymorphisms have been widely used of screening the susceptibility gene in recent years. However, There are relatively few reports about the relationship between the SNPs of HLA-DRB1 and essential hypertension.
Objective
In order to explore the association between HLA-DRB1 gene polymorphism and essential hypertension in the terms of gene level, provide new clues for the genetic mechanism of hypertension, and lay the theoretical basis for clinical treatment of hypertension, rs2308765, rs9269186 and rs3135388 will be slelected to study the relationship between HLA-DRB1 and essential hypertension. And the internal environmental factors of hypertension will be analyzed for the purpose of further exploring the factors affecting the development of hypertension, and providing a reasonable reference for the prevention and control measures.
Methods
The SNPs of HLA-DRB1 were detected by polymerase chain reaction-ligase detection reaction technique in samples from 161 patients with essential hypertension and 167 healthy controls. The Genemapper software was used for the analysis of SNPs data and gene types.
Epidata 3.0 and SPSS13.0 softwares were used to perform analysis. Mean and standard deviance was used to descripe continuous variables. Rate and ratio were used to describe categorical data. T test or Rank test was used to test the differences of continuous variables between case and control.χ2 or Fisher’s exact test were used to test the differences of categorical data between different groups. The goodness-of-fitχ2 test was used for Hardy-Weinberg equilibrium test in case and control. The differences of genetype and allele frequencies between case and control were test byχ2 test. T test or Rank test was used for univariate analysis on the enviromental factors. The multivariable non-conditional logistic regression model was used for the multivariate analysis and the interaction analysis between gene polymorphisms and enviromental factors.
Results
1. 308 samples were investigated, which were divided into two groups. 161 patients belonged to case group, while 147 samples belonged to control group. The essential characteristics of objectives included the following aspects: The average age was 64.6±10.2, and the 60～69 age-group had the highest constituent ratio(35.1%), While the proportion of less than 50 years was the lowest; Male took the most part of samples(92.8%); Most of them were highly educated, about 82.4% were Junior College or College; Han was the main nationality(92.2%). The differences in age, gender, nationality and cultural status were not found between case and control.
2. The results by PCR-LDR indicadated that, Only G/G and G/T genetypes of rs2308765 expressed ; C/C, G/G and C/G genetypes of rs9269186 were found in the research; C/C and T/T genetypes of rs3135388 were found.
3. The genetypes and alleles distributions of rs2308765, rs9269186 and rs3135388 between case and control were accorded with Hardy-Weinberg equilibrium law (P>0.05).
4. The differences of genetype and allele distributions in rs2308765 between case and control were significant (P<0.05). When the mutation was taken as an exposed factor, The OR values of genetype and alleles respectively were 2.774 and 2.675, and the 95% confidence interval of OR didn’t include 1.
5. The significant differences of genetype and allele distributions in rs9269186 and rs3135388 were not found.
6. Between case and control, the low-density lipoprotein showed an edge significace(P=0.059), and BMI, waist circuit, blood sugar, total cholesterol, triglycerides and high-density lipoprotein showed significant differences(P<0.05). The value of BMI, waist circuit, blood sugar, total cholesterol, triglycerides and low-density lipoprotein in case were higher than in control, while the high density lipoprotein value was lower than in control.
7. The multivariable logistic regression showed that high BMI and high triglycerides were risk factors of essential hypertension.
8. The interation annalysis indicated that there was a negtive plus interation between rs2308765 polymorphisms and triglycerides.
Conclusions
1. There was a certain correlation between rs2308765 polymorphisms and essential hypertension, and G/T mutation might be the risk factor of essential hypertension.
2. rs9269186 and rs3135388 polymorphisms did not associate with essential hypertension.
3. The value of BMI, waist circuit, blood sugar, total cholesterol, triglycerides and low-density lipoprotein in case were higher than in control, while the high density lipoprotein value was lower than in control.
4. High BMI and high triglycerides were the risk factors of essential hypertension.
5. There was a negtive plus interation between rs2308765 polymorphisms and triglycerides.
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