Association Of The WNK4 Gene And CYBA Gene Polymorphisms With Essential Hypertension

OBJECTIVE: To explore the polymorphism G1662A of WNK4and the polymorphism C242T of CYBA for association with essentialhypertension; To explore a convenient and rapid method to optimizepolymerase chain reaction (PCR) conditions.METHODS: The genomic DNA was extracted from 135 patientswith essential hypertension (HT) and 135 normotensive (NT) Hansubjects in Hunan. Association analysis was employed. Uniform Designwas applicated to optimize the four factors that may affect the polymerasechain reaction (RCR). The two single nucleotide polymorphisms weregenotyped by polymerase chain reaction-restriction fragment lengthpolymorphism (RCR-RFLP) analysis.RESULTS: (1) The most favorable PCR conditions of WNK4 andCYBA were: annealing temperature 59.8℃and 58.1℃, DNA templatecontent 30ng and 20ng, the concentration of Mg2~+ 1.0 mmol/L and1.5mmol/L respectively. Both the favorable Taq polymerase contents are1.5U. (2) The G1662A genotypic frequency did not deviate fromHardy-Weinberg equilibrium. The frequency of genotype GG was higherthan GA and AA in both groups (HT: 87.40% vs 11.90% vs 0.70%,P＜0.05; NT: 94.10% vs 5.90% vs 0%, P＜0.05).There was no significantlydifference of the distributions of three genotypes (GG, GA, and AA)between the hypertensive and the normotensive groups (chi (2)=3.571,P＞0.05). But the difference of the distributions of T and C alleles wasfound significantly in both groups (chi (2)=4.041, P＜0.05). (3) TheC242T genotypic frequency did not deviate from Hardy-Weinbergequilibrium. The frequency of genotype CC was higher than CT in bothgroups (HT: 83.70% vs 16.30%, P＜0.05; NT: 74.10% vs 25.90%,P＜0.05). The genotype CC was no found. There was no significantlydifference of the distributions of two genotypes (CC and CT) between thehypertensive and the normotensive groups (chi(2)=3.758, P＞0.05), thesame as the distributions of T and C alleles in both groups (chi (2)=3.315,P＞0.05).CONCLUSION: (1) Uniform Design can act as a powerful tool for the optimization of PCR reaction conditions. (2) There was acorrelationship between the WNK4 G1662A polymorphism and essentialhypertension. A allele may be a risk factor of essential hypertension. (3)There was no correlationship between the CYBA C242T polymorphismand essential hypertension.