Choudhuri Subhadip, Chowdhury Imran H, Das Shibali, Dutta Deep, Saha Avijit, Sarkar Rajarshi, Mandal Lakshmi K, Mukherjee Satinath, Bhattacharya Basudev
Department of Biochemistry, Dr. B C Roy Post Graduate Institute of Basic Medical Education and Research (IPGME&R), 244B, A.J.C Bose Road, Kolkata, 700020, India,
Mol Cell Biochem. 2015 Jul;405(1-2):265-79. doi: 10.1007/s11010-015-2417-z. Epub 2015 May 9.
The present study was aimed to investigate the relation between nuclear factor kappa beta (NFκB) activation and downstream up-regulation of vascular endothelial growth factor (VEGF) in diabetic retinopathy (DR). Moreover the study was intended to evaluate the role of VEGF gene single nucleotide polymorphisms (SNPs) in DR occurrence and to investigate the functional relevance of VEGF gene SNPs in terms of VEGF expression in DR. Serum level of VEGF, VEGF R1 (receptor 1), VEGF R 2 (receptor 2) and NFκB (p50/65) activity was measured by enzyme linked immune sorbent assay. Genotyping and allelic composition of different SNPs i.e., rs2010963, rs3025039, rs1570360 and rs 2071559 were investigated by Taqman SNP genotyping assay. VEGF, NFκB p50/p65, and VEGF R1 & R2 gene expressions were quantified by real time quantitative polymerase chain reaction. Increased NFκB p50/p65 activity and expressions were observed in non proliferative diabetic retinopathy (NPDR) and proliferative diabetic retinopathy (PDR) subjects compared to type 2 diabetes mellitus without retinopathy (DNR) group. Significantly elevated levels of serum VEGF and highest VEGF expression were found among PDR subjects compared to DNR or NPDR subjects. CC genotype and C allele of rs2010963 and TT genotype and T allele of rs3025039 were significantly over represented among PDR subjects compared to DNR group. Increased activation of NFκβ in NPDR and PDR subjects might involve increased up regulation of VEGF. VEGF SNPs i.e., rs2010963 C allele and rs3025039 T allele might be associated with PDR occurrence and in turn regulates VEGF expression among PDR subjects.
本研究旨在探讨糖尿病视网膜病变（DR）中核因子κB（NFκB）激活与血管内皮生长因子（VEGF）下游上调之间的关系。此外，该研究旨在评估VEGF基因单核苷酸多态性（SNP）在DR发生中的作用，并从DR中VEGF表达的角度研究VEGF基因SNP的功能相关性。采用酶联免疫吸附测定法测量血清VEGF、VEGF R1（受体1）、VEGF R2（受体2）水平和NFκB（p50/65）活性。通过Taqman SNP基因分型测定法研究不同SNP即rs2010963、rs3025039、rs1570360和rs2071559的基因分型和等位基因组成。通过实时定量聚合酶链反应对VEGF、NFκB p50/p65以及VEGF R1和R2基因表达进行定量。与无视网膜病变的2型糖尿病（DNR）组相比，在非增殖性糖尿病视网膜病变（NPDR）和增殖性糖尿病视网膜病变（PDR）患者中观察到NFκB p50/p65活性和表达增加。与DNR或NPDR患者相比，PDR患者血清VEGF水平显著升高且VEGF表达最高。与DNR组相比，PDR患者中rs2010963的CC基因型和C等位基因以及rs3025039的TT基因型和T等位基因显著过量。NPDR和PDR患者中NFκβ激活增加可能涉及VEGF上调增加。VEGF SNP即rs2010963的C等位基因和rs3025039的T等位基因可能与PDR的发生有关，进而调节PDR患者中的VEGF表达。
