Association Analysis Of ENaC Coding Gene Polymorphisms With Essential Hypertension In Mongolian Population

Objective: To analyze the therapeutic effect of amiloride combined with other antihypertensive drugs targeting epithelial sodium channel(ENaC)on essential hypertension,and to explore the relationship between single nucleotide polymorphisms(SNPs)of ENaC coding gene and the expression level of ENaC protein in urinary exosomes and the development of essential hypertension in Mongolian people in Inner Mongolia.relationship.Methods:(1)A large number of patients with hypertension treated with amiloride were reviewed in the previous data,and those eligible for the randomized control method were screened.Meta-analysis was carried out using Review Manger 5.3 software.(2)Mongolian population obtained from on-site epidemiological surveys conducted in Hohhot,Xilinhot,and Tongliao cities of Inner Mongolia Autonomous Region was used as the study population,and 1062 cases of Mongolian primary hypertension case-control samples were finally included according to the inclusion and exclusion criteria of this study,including 714 cases in the case group and348 cases in the control group.Ten SNPs of ENaC-encoding genes(SCNN1A,SCNN1 B and SCNN1 G genes)were screened in this study,and SNPs were genotyped using multiplex linkage detection reaction(IMLDR).Hardy-Weinberg equilibrium test and association analysis of alleles with hypertension were performed using Haploview software.Association analysis between genotype and hypertension and blood pressure levels was performed using SPSS25.0 software;(3)Urine from subjects was ultra-high-speed centrifuged(200,000 g,2 h),and whether the ultra-high-speed centrifugation products were urinary exosomes was confirmed by nanoparticle tracking analysis(NTA)technique and transmission electron microscopy observation.Urinary exosomes from a case-control population containing 77 Mongolian hypertensive patients and 33 normotensive subjects were subjected to 3D Label Free proteome sequencing to detect the expression of ENaC in urinary exosomes.Results:(1)Meta-analysis showed a positive effect of both amiloride combined with amlodipine and amlodipine combined with telmisartan on the treatment of essential hypertension,and the two dosing regimens did not show significant differences in blood pressure regulation(DBP 82.7±6.5,95% CI [-0.80,0.75],P > 0.05;SBP 131.1±9.1,95% CI[-0.66,1.65],P > 0.05).(2)Association analysis of ENaC-encoding gene polymorphisms with primary hypertension in Mongolians showed that a dominant model at the rs4499238 locus of the SCNN1G gene and a recessive model at the rs4764586 locus of the SCNN1A gene were associated with the development of hypertension in the Mongolian population(P 0.023 and 0.044,respectively).Stratified analysis showed that the variant at the rs876905 locus of the SCNN1A gene was a protective factor for hypertension in a population of smokers(OR = 0.39,95% CI 0.17 to 0.87,P = 0.022);in a normal population,the variant at the rs238547 locus of the SCNN1 B gene was a risk factor for hypertension(OR = 7.27,95% CI 0.85 to 62.10,P =0.027);in men and obese people,the variant at rs4499238 in the SCNN1 G gene was a protective factor for hypertension(men: OR = 0.45,95% CI 0.22 to 0.93,P = 0.034;obese people: OR = 0.31,95% CI 0.14 to 0.70,P = 0.005 The gene polymorphism at the rs5729 locus of the SCNN1 G gene was a risk factor for hypertension in an exercising population(OR = 14.22,95% CI 1.64 to 123.15,P = 0.005).rs4764586 locus of the SCNN1A gene had an effect on diastolic blood pressure levels(P = 0.025).rs61731141 locus The gene polymorphism of the rs61731141 locus was associated with the regulation of systolic blood pressure levels(P = 0.035).(3)3D Label Free proteome sequencing,which showed no detectable expression of ENaC protein in urinary exosomes.Conclusions:(1)Meta-analysis confirmed no significant difference in the effect of two treatment regimens,amiloride combined with amlodipine and temisartan combined with amlodipine,for the treatment of hypertension;(2)at the gene level,the rs4764586 and rs879605 loci of SCNN1A and rs238547 loci of SCNN1 B and the rs4499238 and rs5729 loci of SCNN1 G The locus gene polymorphism of rs5729 was associated with Mongolian essential hypertension,and the rs4764586 and rs61731141 loci of SCNN1A had effects on diastolic and systolic blood pressure,respectively.(3)At the protein level,no expression of ENaC protein was detected in urinary exosomes.