Leptin receptor gene K656N polymorphism is associated with low body fat levels and elevated high-density cholesterol levels in Mexican children and adolescents.

INTRODUCTION: This study evaluated the association between obesity and the K109R, Q223R, and K656N polymorphisms of the leptin receptor (LEPR) locus. Such polymorphisms cause changes in the extracellular extreme of the LEPR gene product and appear to be related to signal transduction toward the cell. SUBJECTS AND METHODS: A total of 128 participants between 6 and 17 years of age from a Mexican Mestizo population were included in the study. Individuals were classified as overweight-obese (n = 76) and normal (n = 52), based on anthropomorphic measurements. The K109R, Q223R, and K656N polymorphisms of the LEPR were determined by the size of restriction fragments obtained from polymorphic fragment amplification (polymerase chain reaction-restriction fragment length polymorphism) obtained from genomic DNA. Allele frequency was compared using the chi-square test. Odds ratio was calculated to determine allele obesity risk factor. RESULTS: Variant allele frequency was 109R = 0.35, 223R = 0.49, and 656N = 0.11 for the K109R, Q223R, and K656N polymorphisms, respectively. No statistically significant association with obesity was found in any of the alleles. The N allele of the K656N polymorphism was associated with nonobesity markers, such as high concentrations of high-density lipoproteins, normal body mass index, less thickness of skinfolds, and body perimeters. None of the alleles studied were shown to be obesity risk factors. CONCLUSIONS: Our results suggest that there is no association between the K109R, Q223R, and K656N polymorphisms of the LEPR gene with obesity, and none of the alleles of the LEPR gene K109R, Q223R, and K656N polymorphisms are an obesity risk factor.

Leptin Levels and Q223R Leptin Receptor Gene Polymorphism in Obese Mexican Young Adults.

INTRODUCTION: The Q223R polymorphism of the leptin receptor (LEPR) gene is one of the most common polymorphisms and it is believed to be associated with a damaged capacity of LEPR signaling and with high circulating leptin levels. METHODS: An observational, cross-sectional, analytical study was carried out in the Autonomous University of Ciudad Juarez, Mexico, where a sample of young adult participants (ranging from 18 to 30 years of age) was obtained. They were classified based on the results of body mass index: non-obese, and overweight/obese. The polymorphic variant was determined by Polymerase Chain Reaction (PCR) from the DNA sample and serum leptin levels were measured by Enzyme-Linked Immuno Sorbent Assay. RESULTS: A total of 159 participants were included (non-obese, n=103; overweight/obese, n=56). Leptin levels were 15.14±12.3 ng/mL in the non-obese group and 26.13±19.0 ng/mL in the overweight/obese group (p≤0.001). The allelic frequencies of the Q and R alleles of the LEPR gene in the studied subjects were as follows: non-obese, Q=0.56, R=0.44; overweight/obese, Q=0.62, R=0.38. The relative risk for the Q/Q genotype was 1.18 (Cl 0.53-2.34), for Q/R was 1.14 (Cl 0.59-2.18) and for R/R was 0.59 (Cl 0.23-1.50). CONCLUSIONS: This study shows that leptin levels are associated with overweight/obesity in Mexican young adults, but this is not related to the presence of the Q223R polymorphism in the LEPR gene, so the underlying mechanisms for a possible disturbance in leptin signaling in obese Mexican young adults await further studies.

WRN polymorphisms affect expression levels of plasminogen activator inhibitor type 1 in cultured fibroblasts.

BACKGROUND: Recessive mutations in WRN gene eliminate WRN protein function (helicase) and cause Werner syndrome. One of the most important clinical features of Werner syndrome patients are the premature onset and accelerated atherosclerosis process. Studies carried out on polymorphic WRN locus have shown that the alleles 1367R and 1074L confer protection for cardiovascular disease. Given that the levels of plasminogen activator inhibitor type 1 (PAI-1) were found to be significantly increased in Werner syndrome patients, is quiet possible that PAI-1 expression could be under regulation of WRN helicase. Therefore the purpose of this work was to evaluate the role of WRN polymorphism in modulating the expression of PAI-1. METHODS: In order to accomplish our aim, an array of primary cultured fibroblasts from normal adult donors was genotyped for polymorphisms of both the WRN and PAI-1 loci. In addition, steady state levels of WRN and PAI-1 were measured by semi-quantitative RT-PCR assays in such cultures. To search for the potential relationship between the lack of WRN protein and PAI-1 expression, heterozygous cultures of fibroblasts (1367RC/1074LF; WRN genotype) were treated with a molecule of interference RNA against WRN messenger RNA (mRNA). RESULTS: We found that, carriers of 1367R and 1074L alleles of WRN shown to have low amounts of PAI-1 in plasma (7.56 +/- 5.02), as compared with carriers of 1367C and 1074F alleles (16.09 +/- 6.03). Moreover, fibroblasts from carriers with these alleles had low expression levels of PAI-1 mRNA. The treatment of heterozygous primary fibroblast cultures (1367RC/1074LF; WRN genotype) with iRNA against WRN mRNA caused PAI-1 overexpression. Treatment with normal PAI-1 inducers (TGFbeta, TNFalpha, or insulin) in these cultures and from those with genotypes 1367CC/1074FF and 1367RR/1074FL resulted in a genotype-dependent PAI-1 expression level. CONCLUSION: Our results suggest that polymorphisms in the WRN gene might have a significant role regulating PAI-1 levels in healthy individuals and "normal states" as well as acute or chronic stress, obesity, aging, acute inflammation, among others, where characteristic high levels of insulin, TNF alpha and TGFbeta, could favor PAI-1 high levels in carriers with polymorphic variants (C and F alleles), beyond the levels reached by carriers with other alleles (R and L alleles).

Influence of serum leptin levels and Q223R leptin receptor polymorphism on clinical characteristic of patients with rheumatoid arthritis from Western Mexico.

OBJECTIVE: The aim of the present study was to evaluate the possible association between the Q223R Leptin receptor (LEPR) polymorphism (A>G; rs1137101) and leptin levels in patients with rheumatoid arthritis (RA) from Western Mexico. METHODS: A cross-sectional study was performed with 70 RA patients and 74 controls subject (CS). Disease activity was evaluated using DAS28 score, the Q223R LEPR polymorphism was determined by the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and serum leptin levels, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and rheumatoid factor (RF) were quantified. RESULTS: RA patients had significant high serum leptin levels compared with CS; leptin levels correlated strongly with body composition measures, but not with inflammatory markers, disease evolution, and activity. The genotype and allele frequencies of the Q223R LEPR polymorphism were not associated with RA. Similarly, leptin levels did not differ between Q223R LEPR genotypes. CONCLUSION: The LEPR Q223R polymorphism was not associated with RA risk in patients from Mexican population, even though high levels of serum leptin were present and these could explain the low weight observed in RA patients when they were compared to control subjects. However, the serum leptin levels did not correlate with inflammatory markers, severity and disease evolution.

Forskolin suppresses delayed-rectifier K+ currents and enhances spike frequency-dependent adaptation of sympathetic neurons.

In signal transduction research natural or synthetic molecules are commonly used to target a great variety of signaling proteins. For instance, forskolin, a diterpene activator of adenylate cyclase, has been widely used in cellular preparations to increase the intracellular cAMP level. However, it has been shown that forskolin directly inhibits some cloned K+ channels, which in excitable cells set up the resting membrane potential, the shape of action potential and regulate repetitive firing. Despite the growing evidence indicating that K+ channels are blocked by forskolin, there are no studies yet assessing the impact of this mechanism of action on neuron excitability and firing patterns. In sympathetic neurons, we find that forskolin and its derivative 1,9-Dideoxyforskolin, reversibly suppress the delayed rectifier K+ current (IKV). Besides, forskolin reduced the spike afterhyperpolarization and enhanced the spike frequency-dependent adaptation. Given that IKV is mostly generated by Kv2.1 channels, HEK-293 cells were transfected with cDNA encoding for the Kv2.1 α subunit, to characterize the mechanism of forskolin action. Both drugs reversible suppressed the Kv2.1-mediated K+ currents. Forskolin inhibited Kv2.1 currents and IKV with an IC50 of ~32 µM and ~24 µM, respectively. Besides, the drug induced an apparent current inactivation and slowed-down current deactivation. We suggest that forskolin reduces the excitability of sympathetic neurons by enhancing the spike frequency-dependent adaptation, partially through a direct block of their native Kv2.1 channels.