Abstract
BackgroundSeveral studies have shown overexpression of leptin in microarray experiments in pre-eclampsia (PE) and in hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome. We decided to study four leptin receptor (LEPR) SNP polymorphisms in HELLP syndrome patients by using quantitative real-time PCR and melting curve analysis.MethodsDNA was isolated from blood samples from 83 normotensive pregnant women and 75 HELLP syndrome patients. Four SNPs, LEPR c.326A>G (K109), LEPR c.668A>G (Q223R), LEPR c.1968G>C (K656N) and LEPR c.3024A>G (S1008) were determined by quantitative real-time PCR and melting curve analysis. Investigators were blinded to clinical outcomes.ResultsLEPR c.326A>G, LEPR c.668A>G, LEPR c.1968G>C and LEPR c.3024A>G allele, genotype and haplotype polymorphisms were not different in HELLP syndrome patients and normotensive healthy pregnants. There were strong linkage disequilibrium (LD) between loci c.326A>G and c.6687A>G (D' = 0.974), and c.668A>G and c.1968G>C (D' = 0.934), and c.326A>G and c.1968G>C (D' = 0.885), and c.1968G>C and c.3024A>G (D' = 1.0). However, linkages of c.3024A>G with c.668A>G (D' = 0.111) and c.326A>G (D' = 0.398) were weak. The Hardy-Weinberg equilibrium was observed for all polymorphisms. However the LEPR c.326A>G AG genotype was twice more frequent and the (AG AG GG AG) haplotype was three times more frequent in HELLP syndrome patients. The introduced quantitative real-time PCR combined with melting curve analysis is a fast and reliable method for the determination of LEPR SNPs.ConclusionAlthough certain LEPR haplotypes are more frequent in HELLP syndrome, we conclude that there is no compelling evidence that the four studied LEPR SNP polymorphisms associated with the development of HELLP syndrome.
Highlights
Several studies have shown overexpression of leptin in microarray experiments in pre-eclampsia (PE) and in hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome
The human leptin receptor is a member of class 1 cytokine receptor family, which was identified in the brain [1]
PCR parameters were optimized for the four primerprobe system to obtain recognizable Tm differences among the different alleles for detection of leptin receptor (LEPR) SNPs (Figure 1)
Summary
Several studies have shown overexpression of leptin in microarray experiments in pre-eclampsia (PE) and in hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome. Leptin regulates the food intake and energy metabolism, it was found that beside that, it stimulates the proliferation of various cell types and it is considered to be a new growth factor. It seems to have effect on immunity, angiogenesis, reproduction, and in the regulation of the blood pressure [10,11]. Leptin acts synergistically with VEGF (vascular endothelial growth factor) and fibroblast growth factor 2 (FGF2) to promote angiogenesis [13] It has effect on the expression of several genes involved in the angiogenesis (MMP-2 and MMP-9) [14,15]. Leptin was implicated in the pathogenesis of several disease including essential hypertension and pre-eclampsia (PE) [16,17]
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