The Relationship between α-adducin Gene Polymorphism and Essential Hypertension
|School||Hebei Medical University|
|Keywords||Essential hypertension α- adducin Gene polymorphism|
Objective: essential hypertension polygenic disease is caused by a genetic and environmental factors, racial differences, explicit hysteresis and heterogeneity characteristics. Adducin by α, β, γ3 of species subunit heterodimer composed or heterologous to the tetramer, profiled elements in the erythrocytes, brain, lung, kidney, liver, and other organizations have been detected. Adduction protein subunits with similar structures can be divided into the the amino terminal spherical head, neck and carboxy-terminal tail. By Ca2 / calmodulin protein kinase C and protein kinase A regulation of α-adducin involved in cellular signal transduction. In addition, α-adducin interaction of other ingredients (such as spectrin and actin, etc.) and a variety of membrane proteins (e.g., ion channels, exchanging body and an ion pump, etc.) with the membrane skeleton, to participate in the cell membrane ion transport, in particular, with a variety of membrane Na transporters, Na-H exchange, Na-Li antiporter and sodium pump activity is closely related to the Na-K-Cl-cotransporter system, thereby affecting blood pressure. Human α-adducin targeted at the 4th chromosome (4p16.3), the the 10 exon G614T single nucleotide polymorphisms, resulting in 460 glycine (Gly) encoding the protein tryptophan (Trp ) replacement. In this experiment, the Han population of north China as the research object, using PCR-RFLP and DNA sequencing methods, partial DNA sequence analysis of exon 10 outside of the α-adducin gene, further clear in northern China Han α-adduction protein gene Gly460Trp by polymorphism with essential hypertension. Methods: 160 cases of patients with essential hypertension in Northern Han and 151 normotensives study, blood pressure was measured using a mercury sphygmomanometer, the corrected measurement upper arm blood pressure 15 minutes before measuring blood pressure in patients with non-smoking, backed by sit-in 5 minutes; measured before enrollment the unusual 2011 3 BP, taking the average; blood pressure measured in the standard state, i.e. the same time each day, the ipsilateral arm, sphygmomanometer using the same, and completed by the same measurement. The inclusion criteria for the prevention and control of hypertension in China Guide 2005: have a clear history of essential hypertension and antihypertensive medication; were not taking anti-hypertensive drugs, and unusual day 2 measured systolic blood pressure ≥ 140mmHg and / or diastolic blood pressure ≥ 90mmHg, exclude secondary hypertension, diabetes, coronary heart disease, hyperlipidemia, severe liver and kidney function impaired. Inclusion criteria were selected to meet the above essential hypertension and no family history of hypertension, to exclude secondary hypertension, diabetes, coronary heart disease, hyperlipidemia, severe liver and kidney dysfunction, the study selected between unrelated. General registration each study clinical data and detect biochemical indicators, including: gender, age, family history, and systolic blood pressure (SBP), diastolic blood pressure (DBP), triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C). All of the subjects to be fasting more than 12 hours in the early morning to take seats blood collection cubital vein. Each blood sample preprocessing corresponding biochemical detection requirements the: centrifugal separation supernatant at -80 ℃ ultra-low temperature refrigerator until the specimen collection complete determination of the same batch, to reduce the error in the batch. Another smoke 2ml venous blood containing anticoagulant vacuum pumping vascular -80 ℃ ultra-low temperature refrigerator, mix well placed for the extraction of genomic DNA. Using the traditional phenol / chloroform extraction method, genomic DNA was extracted from peripheral blood leukocytes by polymerase chain reaction (PCR-RFLP) method to detect α-adducin 10 exon gene G614T (G for guanine, T thymine) gene polymorphism, reagents are: phosphate buffer solution (PBS), proteinase K (Pro Nase in K) Extraction Buffer, Tris-saturated phenol, chloroform, isoamyl alcohol, and ethanol. Other experimental reagents and instruments: the polymerase chain reaction primers (Shanghai Biological Engineering Technology \u0026 Services Co., Ltd.), dNTPs (Beijing race Parkson Gene Technology Co., Ltd.), DNA marker the RedTaq DNA polymerase (Beijing Parkson race gene technology) 12% polyacrylamide gel (PAG), restriction endonuclease (New England Bio-Technology Co., Ltd.), British Hybaid Express automatic instrument, USA Kodak gel imaging system. All data using SAS 6.12 statistical software for data analysis. Gene frequencies by gene counting method, the allele frequency ratio using the chi-square test, measurement data are expressed as mean ± standard deviation, count data frequency. Hardy-Weinberg equilibrium test, group comparison between genotype and allele frequencies of fourfold table χ2 test; the different genotype blood pressure and indicators of differences in the two independent samples t-test. P lt; 0.05 was considered statistically significant. Results: 1. General clinical data and biochemical indicators of comparison: between the two groups in the average age of TC, LDL-C, HDL-C, TG no significant difference (P gt; 0.05); essential hypertension group and normal control group compared to SBP, DBP water were higher than those in the latter have a significant difference (P lt; 0.05), and therefore comparable. The chi-square test of each group genotype distribution with Hardy-Weinberg genetic equilibrium, the representative of the study population. Adducin 3.α-gene Gly460Trp polymorphism in the EH group and the control group of three genotypes than the difference was not statistically significant (P gt; 0.05); allele frequency difference was not statistically significant (P gt; 0.05). 4. GlyTrp, and TrpTrp genotype systolic blood pressure levels higher than GlyGly genotype patients (P lt; 0.05); GlyTrp genotypes in patients with diastolic blood pressure level to higher than GlyGly and TrpTrp genotype (P lt; 0.05). Conclusion: α-adducin gene Gly460Trp polymorphism in northern China Han population, but the gene polymorphism with essential hypertension found no significant correlation; carry α-adducin 460Trp allele in patients with contraction higher pressure level; GlyTrp genotype had higher diastolic blood pressure levels.