Mehranpour P, Wang S S, Blanco R R, Li W, Song Q, Lassègue B, Dikalov S I, Austin H, Zafari A M
Emory University School of Medicine, Division of Cardiology, Atlanta, GA 30322, USA.
Cardiovasc Hematol Agents Med Chem. 2009 Jul;7(3):251-9. doi: 10.2174/187152509789105417.
Single nucleotide polymorphisms (SNP) in the CYBA gene encoding p22(phox) have been associated with respiratory burst and cardiovascular phenotypes. We previously reported a reduced phagocytic respiratory burst activity in healthy adults with the C242T SNP, but found no correlation between CYBA SNPs and coronary artery disease (CAD) phenotype. Using lymphoblastoid cells, we hypothesized that CYBA SNPs affect enzyme activity in patients with cardiovascular disease (CVD), but would not be associated with angiographic severity of CAD due to confounding by risk factors. We established lymphoblastoid cell lines from patients with CVD and genotyped the study cohort for CYBA SNPs and phenotyped each subject's coronary angiogram for CAD severity. As quantified by electron spin resonance, superoxide production in picomoles per 10(6) resting lymphoblastoid cells per minute for the CC, CT, and TT genotypes of the C242T SNP were 16.2+/-1.4, n=70, 11.9+/-0.7, n=87, and 11.9+/-1.5, n=28, respectively (P=0.002). The -930(A/G) and A640G SNPs did not affect superoxide production (P > 0.2). Expression of p22(phox) was not affected as determined by real-time RT-PCR and Western blot analysis. The C242T CYBA SNP is associated with altered NADPH oxidase activity in lymphoblastoid cells of patients with CVD. By reducing the influence of confounding environmental factors, lymphoblastoid cell lines could serve as a tool to assess direct genotype/phenotype interactions of candidate genes known to affect atherosclerosis.
编码p22(phox)的CYBA基因中的单核苷酸多态性(SNP)与呼吸爆发和心血管表型相关。我们之前报道，携带C242T SNP的健康成年人吞噬性呼吸爆发活性降低，但未发现CYBA SNPs与冠状动脉疾病(CAD)表型之间存在相关性。我们使用淋巴母细胞系推测，CYBA SNPs会影响心血管疾病(CVD)患者的酶活性，但由于危险因素的混杂作用，不会与CAD的血管造影严重程度相关。我们建立了CVD患者的淋巴母细胞系，对研究队列进行CYBA SNPs基因分型，并对每个受试者的冠状动脉造影进行CAD严重程度的表型分析。通过电子自旋共振定量分析，C242T SNP的CC、CT和TT基因型在每分钟每10(6)个静息淋巴母细胞中产生的超氧化物的皮摩尔数分别为16.2±1.4(n = 70)、11.9±0.7(n = 87)和11.9±1.5(n = 28)(P = 0.002)。-930(A/G)和A640G SNPs不影响超氧化物的产生(P>0.2)。通过实时逆转录PCR和蛋白质印迹分析确定，p22(phox)的表达不受影响。C242T CYBA SNP与CVD患者淋巴母细胞中NADPH氧化酶活性的改变有关。通过减少混杂环境因素的影响，淋巴母细胞系可作为一种工具，用于评估已知影响动脉粥样硬化的候选基因的直接基因型/表型相互作用。
