Abstract
BackgroundP-selectin, encoded by SELP, has been implicated as an important molecule in the development of arterial stiffness, consequently leading to vascular complications in T2DM. SELP polymorphisms and increased levels of soluble P-selectin (sP-selectin) have been shown to be associated with several inflammatory diseases. The present work was designed to assess nine putative functional non-coding SELP variants in relation to sP-selectin levels and arterial stiffness in T2DM.MethodsThe genetic distribution of rs3917655, rs3917657, rs3917739, rs2235302, rs3917843 was determined by restriction fragment length polymorphism–polymerase chain reaction (RFLP-PCR). Genotyping of rs3917779 was performed by tetra primer amplification-refractory mutation system (ARMS)- PCR. Three SNPs i.e. rs3917853, rs3917854, rs3917855 were genotyped by Sanger sequencing. Construction of haplotypes was performed using PHASE software. The data thus obtained was analyzed by appropriate statistical tools.ResultsTwo non-coding variants i.e. rs3917657 and rs3917854 of SELP were found to be associated with 2 and 1.7 -fold risk of disease development respectively. However, one non-coding variant rs2235302 was found to provide protection against disease development. Furthermore, variant allele of rs3917854 in T2DM patients was found to be associated with 2.07-fold very high vascular risk. Non-coding haplotype GCAGGCCGC was conferring 4.14-fold risk of disease development. Furthermore, overall sP-selectin levels were higher in T2DM patients when segregated according to genotypes as well as haplotypes. Significant genotype- phenotype correlation was observed for rs3917655 as well as rs3917739 variant in patients and for rs3917854 in controls. In vascular risk categories, a significant genotype- phenotype correlation was observed for rs3917655 and rs2235302. Furthermore, patients with CCGGGCCGC haplotype in high risk category were observed with higher levels of sP-selectin as compared to other haplotypes (p < 0.05).ConclusionsNon-coding SELP variants may significantly modulate sP-selectin levels, vascular risk and T2DM susceptibility.
Highlights
P-selectin, encoded by SELP, has been implicated as an important molecule in the development of arterial stiffness, leading to vascular complications in Type 2 diabetes mellitus (T2DM)
Genotyping of SELP variants A total of nine single-nucleotide polymorphisms (SNPs) selected on the basis of in silico analyses were genotyped by various methods including Restriction fragment length polymorphism (RFLP)-Polymerase chain reaction (PCR), amplification-refractory mutation system (ARMS)-PCR and Sanger sequencing
The representative agarose gels showing PCR products and restriction digestion products as well as electropherograms of representative samples for all the studied variants are given in supplementary figure 1–7
Summary
P-selectin, encoded by SELP, has been implicated as an important molecule in the development of arterial stiffness, leading to vascular complications in T2DM. Binding of P-selectin to its ligands mediate initial steps of adhesion cascade i.e. tethering and rolling [21, 22]. This interaction further results into proteolytic shedding of P-selectin in circulation as soluble P-selectin (sP-selectin), which is documented as marker of endothelial dysfunction and platelet hyperactivity [23,24,25,26,27]. Studies have suggested a significant association of raised sP-selectin levels with atherosclerotic vascular complications including coronary heart disease (CHD), CAD and MI in T2DM [26, 28,29,30,31,32]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
