| IntroductionEssential hypertension (EH) is the most common cardiovascular disease, however the pathogenesis of EH have not yet been reported. EH is a complex disease that results from genetic factors and environmental factors. Some SNPs and their certain combinations of these genes may be one of the key factors of liability to this kind of diseases. It is hopeful to obtain breakthroughs in studying diseases such as hypertension, diabetes and cancer by the research of SNP association analysis of candidate genes related to the diseases or playing some roles on regulating pathways of the diseases, - adrenergic receptor ( - AR) gene plays an important role on regulating blood pressure and the occurrence of cardiovascular diseases and it is an important candidate gene of EH. PCR - RFLP was used to detect the distribution frequency of the three subtypes of - AR gene family in northeast China with high occurrence frequency of hypertension.The case - control study was carried out between hypertensives and normosives to analyze the association of the polymorphisms with essential hypertension so as to discuss the significance of the four polymorphisms in hypertension in northeast China. Meanwhile 1 - AR eukaryotic expressing vectors were constructed to study primarily the influence of different cSNP on gene function through cell transfection and detection of AC activity.Materials and MethodsMaterials1. Reagents for cell culture2. COS7 cell line3. Molecular biology reagents of gene cloning4. Reagents for cell transfection5. Radioimmunoassay Kit MethodsPCR - RFLP was used to detect the genotypes of hypertensives and nomo-tensives in the four cSNPs of the three subtypes of - AR gene family. The case - control study was carried out between hypertensives and normosives to analyze the association of the polymorphisms with essential hypertension, in which \ test was used. Meanwhile some indexes were tested and analyzed with t - test. Eukaryotic recombined expression vector, pcDNA3. 1 - - AR was constructed and then transfected COS? cell by lipofection. Radioimmunoassay was performed to detect the concentration of cAMP in the transfected cells to examine AC activity indirectly.ResultsThe frequency of homozygote for the Gly389 allele of the 1 - AR Arg389Gly was higher in hypertensives than in normotensives and the frequency of homozygote for the Ser49 allele of the 1 - AR Gly49Ser was significantly higher in hypertensives than in normotensives. No significantly statistical differences were found in the frequencies of Argl6Gly of 2 - AR and Trp64Arg of 3 - AR between hypertensives and normotensives. The four cSNPs are not associated with the indexes tested. The three pcDNA3. 1 - 1 - AR expressing vector with three different genotype combinations were constructed successfully. After cell transfection, we found that the concentration of cAMP in the cells transfected with homozygote for Gly389 combined vector was higher than those with homozygote for Arg389 combined vector if the agonist is not used, the result is adverse if the agonist is used. The concentration of cAMP in the cells transfected with homozygote for Ser49 combined vector is higher than those with homozygote for Gly49 combined vector whether the agonist is used or not.Conclusion1. Arg389Gly and Ser49Gly polymorphism of β1 - AR were involved in the pathogenesis of hypertension. Individuals homozygous for Gly389 allele and Ser49 of the β1 - AR are at increased risk to develop hypertension. Argl6Gly polymorphism of β2 - AR and Trp64Arg polymorphism of β3 - AR are not associated with hypertension. The four cSNPs are not associated with the indexes tested.2. The AC activity of cells homozygous for Gly389 allele is higher if the agonist is not used, the result is adverse if the agonist is used. And that of Ser49 allele is higher whether the agonist is used or not. .
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