Ambreen Fareeha, Ismail Muhammad, Qureshi Irfan Zia
Laboratory of Animal and Human Physiology, Quaid-i-Azam University, Islamabad, Pakistan.
Institute of Biomedical and Genetic Engineering, Islamabad, Pakistan.
Mol Vis. 2015 Aug 29;21:985-99. eCollection 2015.
To study the association of serum levels of inflammatory mediators and angiogenic factors with genetic polymorphism in Pakistani age-related macular degeneration (AMD) patients.
This was a cross-sectional and case-control study that included 90 AMD patients diagnosed through slit-lamp examination, fundoscopy, and ocular coherence tomography. For reference and comparison purposes, 100 healthy age-matched subjects (controls) were also recruited. IL-6, IL-8, VEGF, and CRP levels were estimated in the serum samples of patients and control subjects. Using restriction fragment length polymorphism, single nucleotide polymorphisms were studied in IL-6 (rs1800795, rs1800796, rs1800797), IL-8 (rs4073, rs2227306, rs2227543), VEGF (rs3025039, rs699947), and CRP genes (rs1205, rs1130864). Since the data were obtained from a sample population, the Box-Cox transformation algorithm was applied to reduce heterogeneity of error. Multivariate analyses of variance (M-ANOVA) were applied on the transformed data to investigate the association of serum levels of IL-6, IL-8, VEGF, and CRP with AMD. Genotype and allele frequencies were compared through χ(2) tests applying Hardy-Weinberg equilibrium. The serum concentrations of IL-6 and IL-8, VEGF, and CRP between homozygotes and heterozygotes were compared through one-way ANOVA. Significance level was p<0.05.
Compared to control subjects, serum IL-6 (p<0.0001), IL-8 (p<0.0001), VEGF (p<0.0001), and CRP (p<0.0001) levels were significantly elevated in the AMD patients. For rs1800795, patients with the GG genotype showed significantly raised levels of IL-6 compared to those with GC and CC genotypes (p<0.0001). Serum IL-8 levels were significantly higher in patients with the GG genotype compared to the GC and CC genotypes for the single nucleotide polymorphism (SNP) rs2227543 (p<0.002). Similarly, significantly higher VEGF levels were detected for genotype TT for rs3025039 SNP (p<0.038). However, no significant alteration in serum CRP levels was detected in hetero- or homozygotes for rs1205 and rs1130864 SNPs.
Serum IL-6, IL-8, and VEGF levels are substantially increased in AMD, and the levels coincide with polymorphism in the respective gene. No such relationship appears to exist with regard to SNPs of CRP.
研究巴基斯坦年龄相关性黄斑变性（AMD）患者血清炎症介质和血管生成因子水平与基因多态性之间的关联。
这是一项横断面病例对照研究，纳入了90例经裂隙灯检查、眼底镜检查和光学相干断层扫描诊断的AMD患者。为作参考和比较，还招募了100名年龄匹配的健康受试者（对照组）。检测了患者和对照受试者血清样本中白细胞介素-6（IL-6）、白细胞介素-8（IL-8）、血管内皮生长因子（VEGF）和C反应蛋白（CRP）的水平。采用限制性片段长度多态性方法，对IL-6（rs1800795、rs1800796、rs1800797）、IL-8（rs4073、rs2227306、rs2227543）、VEGF（rs3025039、rs699947）和CRP基因（rs1205、rs1130864）的单核苷酸多态性进行研究。由于数据来自样本群体，应用Box-Cox变换算法以减少误差的异质性。对转换后的数据进行多变量方差分析（M-ANOVA），以研究血清IL-6、IL-8、VEGF和CRP水平与AMD的关联。通过应用哈迪-温伯格平衡的χ²检验比较基因型和等位基因频率。通过单因素方差分析比较纯合子和杂合子之间IL-6、IL-8、VEGF和CRP的血清浓度。显著性水平为p<0.05。
与对照受试者相比，AMD患者血清IL-6（p<0.0001）、IL-8（p<0.0001）、VEGF（p<0.0001）和CRP（p<0.0001）水平显著升高。对于rs1800795，GG基因型患者的IL-6水平显著高于GC和CC基因型患者（p<0.0001）。对于单核苷酸多态性（SNP）rs2227543，GG基因型患者的血清IL-8水平显著高于GC和CC基因型患者（p<0.002）。同样，对于rs3025039 SNP的TT基因型，检测到VEGF水平显著更高（p<0.038）。然而，对于rs1205和rs1130864 SNPs的杂合子或纯合子，未检测到血清CRP水平有显著变化。
AMD患者血清IL-6、IL-8和VEGF水平显著升高，且这些水平与各自基因的多态性相符。CRP的单核苷酸多态性似乎不存在这种关系。
