诱导型一氧化氮合酶基因多态性与川崎病并发冠状动脉病变易感性研究

Correlation between inducible nitric oxide synthase gene polymorphism and affectability of Kawasaki disease coincidenced coronary artery lesion

目的:探讨诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)基因多态性与川崎病(Kawasaki disease,KD)并发冠状动脉病变(coronary artery lesion,CAL)的相关性。方法:将本院收治的146例KD患儿作为KD组,同期健康体检儿童119例作为健康对照组。KD组中合并CAL(CAL组)79例,未合并CAL(none coronary artery lesion,NCAL组)67例。利用基质辅助激光解吸电离飞行时间质谱法检测iNOS基因-1026C/A和+2087G/A 2个单核苷酸多态性(single nucleotide polymorphisms,SNP)位点基因型。应用酶联免疫吸附实验(ELISA)双抗体夹心法检测血浆iNOS水平,硝酸还原法检测血浆NO水平。比较各组间血浆iNOS、NO表达水平,分析2个SNP位点多态性与KD并发CAL相关性。结果:健康对照组血浆iNOS水平(48.02±31.46)U/L明显低于NCAL组(81.46±41.32)U/L和CAL组(90.29±47.68)U/L(P=0.000、0.000)。血浆NO水平组间比较差异无统计学意义(F=3.003,P=0.052),iNOS基因+2087G/A位点A等位基因及GA+AA基因型频率在CAL组明显低于NCAL组(P=0.010、0.013),与G等位基因及GG基因型相比,A等位基因及GA+AA基因型均能显著降低KD并发CAL发生风险[OR(95%CI)=0.41(0.21-0.82),0.42(0.19-0.92);P=0.010,0.029]。-1026C/A与+2087G/A 2位点构建的-1026C/+2087A单倍型频率分布在CAL组与NCAL组间差异有统计学意义(P=0.010),-1026C/+2087A单倍型携带者发生KD并发CAL的风险显著降低[OR(95%CI)=0.45(0.22-0.91),P=0.030]。结论:血浆iNOS水平在KD及其CAL的发生过程中可能起一定作用。iNOS基因+2087G/A多态性和-1026C/+2087A单倍型与KD并发CAL密切相关,其中+2087A等位基因可能为减少KD并发CAL发生风险的保护基因,其作用机制可能通过降低血浆iNOS水平,减少NO产量实现。

Objective：To investigate the correlation between inducible nitric oxide synthase（iNOS） gene polymorphism and the development of coronary artery lesion（CAL） in Kawasaki disease（KD）. Methods：Totally 146 patients with KD in the Children＇s Hospital in Chongqing Medical University were included as KD group and 119 healthy children at the same period as control group. There were 79 KD patients with CAL as CAL group and 67 patients without CAL as NCAL group. SNPs of iNOS gene loci -1026C/A and ＋2087G/A were genotyped by using MassARRAY-IPLEX technology and matrix-assisted laser desorption ionization time of flight inass spectrometry platform（MALDI-TOF-MS）. Relation between gene polymorphisms and the development of CAL in KD were analyzed. Plasma iNOS level was determined by ELISA,plasma NO level was measured by nitric acid reductase and expression levels were compared between groups. Results：Plasma iNOS level was significantly lower in control group （48.02 ±31.46） U/L than that in CAL group （90.29 ± 47.68） U/L and NCAL group（81.46±41.32） U/L（P=0.000,0.000）. Plasma NO level was not significantly different between groups（F=3.003,P=0.052）. Frequencies of the A minor allele and GA＋AA genotypes of ＋2087G/A were higher in CAL group than in NCAL group （P=0.010,0.013） ;both can significantly reduce the risk of CAL compared with G allele and GG genotype[OR （95%CI）=0.41 （0.21-0.82）,0.42（0.19-0.92） ,P=0.010,0.029]. Frequencies of haplotype -1026C/＋2087A were different between CAL group and NCAL group（P=0.010）. -1026C/＋2087A haplotype was associated with a significantly decreased risk of CAL[OR （95% CI）=0.45 （0.22-0.91）,P=0.030]. Conclusions：Results suggested that plasma iNOS is important in the development of vasculitis and CAL in KD. iNOS gene ＋2087G/A polymorphism and its -1026C/＋2087A haplotype are associated with the development of CAL in KD. ＋2087A allele may significantly reduce the risk of CAL by lowering plasma iNOS level and reducing NO production.