The relationship of neutrophil-to-lymphocyte ratio and platelet-to-lymphocyte ratio with bone mineral density in adolescent girls suffering from anorexia nervosa

Introduction: Inflammation is supposed to be one of the factors contributing to decreased bone mineral density (BMD) in anorexia nervosa (AN). The neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) are simple and cost-effective inflammatory markers, well documented as indicators of postmenopausal osteoporosis. This study aimed to assess the relationships between these ratios and BMD in girls with AN. Material and methods: The electronic records of 73 girls hospitalized for AN were analysed retrospectively. The age range of the study group was 12.56–17.67 years. BMD was assessed by dual-energy X-ray absorptiometry (DXA) and expressed as Z-scores according to lumbar spine (s-BMD) and total body (TB-BMD) sites. NLR and PLR were calculated according to complete blood count results. Patients were divided into 2 subgroups with parallel analyses — according to the TB-BMD criterion and the s-BMD criterion: normal (Z-score > –2.0, n = 63) and low s-BMD subgroup (Z-score ≤ –2.0, n = 10), and normal (Z-score > –2.0, n = 45) and low TB-BMD subgroup (Z-score ≤ –2.0, n = 28). Results: In the low s-BMD subgroup a tendency to an increase of mean NLR, PLR, and WBC values was observed. Respective BMD Z-score values correlated significantly and negatively with PLR in the low s-BMD (R = –0.892, p < 0.001) and normal TB-BMD (R = –0.451, p = 0.002) subgroups, while with NLR only in the normal TB-BMD subgroup (R = –0.685, p < 0.001). In the low s-BMD subgroup the PLR was shown to be a significant and independent predictor of s-BMD (β = –0.881, p < 0.001). The PLR contributed to 77.6% of the s-BMD Z-score variability (R2 = 0.776, p < 0.001). In the normal TB-BMD subgroup, the PLR and NLR levels were significant and independent predictors of TB-BMD (β = –0.352, p = 0.004; β = –0.450, p = 0.001; β = –0.339, p = 0.005, respectively) and explained 44.4% of TB-BMD Z-score variability (R2 = 0.444, p < 0.001). Conclusions: These results indicate that there might be a relationship between bone mass loss and inflammation expressed as NLR and PLR in adolescent girls suffering from AN. These connections seem to be dependent on the examined skeletal area. NLR and PLR, which are common indicators of morbidity and mortality in many malignancies and inflammatory chronic diseases, can also be useful in the evaluation of bone condition in adolescent females with AN. However, there is a need for further investigation in this field.

Evaluation of the frequency of ADIPOQ c.45 T>G and ADIPOQ c.276 G>T polymorphisms in adiponectin coding gene in girls with anorexia nervosa

Introduction: Anorexia nervosa (AN) is a serious chronic psychosomatic disorder, the essence of which are attempts by the sufferer to obtain a slim silhouette by deliberate weight loss (restrictive diet, strenuous physical exercise, provoking vomiting). The aetiology of this disorder is multifactorial. Genetic factors that influence the predisposition to AN have been sought. A broad meta-analysis points to a strong genetic correlation between AN and insulin resistance. Adiponectin (ADIPO) increases insulin sensitivity. In our pilot study we demonstrated that the TT genotype in locus ADIPOQ c.276 G>T of the ADIPO gene and a higher concentration of ADIPO in blood serum occurred significantly more frequently in 68 girls suffering from AN than in 38 healthy girls. The objective of this study was to evaluate the frequency of the occurrence of ADIPOQ c.45 T>G and ADIPOQ c.276 G>T in the ADIPO gene in a larger cohort of girls with AN and healthy girls, as well as an analysis of correlations between variants of the aforementioned polymorphisms and the levels of ADIPO in blood serum. Material and methods: The study covered 472 girls (age: 11–19 years): 308 with the restrictive form of AN (AN) and 164 healthy girls (C). The level of ADIPO in blood serum was determined by means of the ELISA method on a Bio-Vendor, LLC (Asheville, North Carolina, USA). The DNA isolation was carried out by means of Genomic Mini AX BLOOD (SPIN). The PCR reaction was carried out in a ThermoCycle T100 thermocycler. 80–150 ng of the studied DNA and relevant F and R starters were added to the reaction mixture. The reaction products were subjected to digestion by restriction enzymes and separated on agarose gels (RFLP). Results: The distribution of genotypes in the polymorphic site ADP c.45 of the ADIPO gene and ADP c.276 was similar in both groups. In both groups the T allele was most frequent in locus ADIPOQ c.45 and the G allele in locus ADIPOQ c.276. In all the study subjects collectively (AN and C) a statistically significant negative correlation between the levels of ADIPO in blood serum on one hand and body weight (r = –0.46; p < 0.0001) and BMI (r = –0.67; p < 0.0001) on the other was demonstrated. Exclusively in the AN group a significant correlation between the level of ADIPO in blood and the distribution of TG, TT, and GG alleles in loci ADIPOQ c.45 and ADIPOQ c.276 was demonstrated (p = 0.0052 and p < 0.0001, respectively). Conclusions: The genotype in loci ADIPOQ c.45 and ADIPOQ c.276 of the ADIPO gene seems to have no effect on the predisposition to AN. Girls suffering from AN with the TT genotype in loci ADIPOQ c.45 and ADIPOQ c. 276 may demonstrate higher insulin sensitivity because they have significantly higher levels of ADIPO than girls suffering from AN with other genotypes. This may be suggestive of their better adaptation to the state of malnutrition, and it has a potential effect on treatment results

Evaluation of the frequency of RETN c.62G>A and RETN c.-180C>G polymorphisms in the resistin coding gene in girls with anorexia nervosa

Introduction: Anorexia nervosa (AN) is a serious psychosomatic syndrome, classified as an eating disorder. AN patients strive to lose weight below the normal limits defined for a specific age and height, achieving their goal even at the expense of extreme emaciation. AN has a multifactorial aetiology. Genetic factors are believed to be significant in the predisposition to the development of AN. In girls suffering from AN significantly lower levels of resistin (RES) in blood serum are observed as compared to healthy girls. These differences may lead to a thesis that functional genetic polymorphisms in RES coding genes can be responsible for this phenomenon. In our pilot study we demonstrated significant differences in the distribution of genotypes in the locus RETN c.-180C>G of the RES gene in 67 girls with AN and 38 healthy girls. It seems reasonable to compare the frequency of polymorphisms of RETN c.62G>A and RETN c.-180C>G in the RES gene in girls with AN and in healthy subjects in a bigger cohort and to analyse correlations between individual variants of the polymorphisms referred to above and the RES levels in blood plasma. Material and methods: The study covered 308 girls with the restrictive form of AN (AN) and 164 healthy girls (C) (aged 11–19 years). The RES levels in blood serum were determined by means of the ELISA method on a Bio-Vendor machine from LLC (Asheville, North Carolina, USA). The DNA isolation was carried out by means of Genomic Mini AX BLOOD (SPIN). The PCR reaction was carried out on a ThermoCycle T100 thermocycler. 80–150 ng of the studied DNA and relevant F and R starters were added to the reaction mixture. The reaction products were subjected to digestion by restriction enzymes and separated on agarose gels (RFLP). Results: The average RES level in blood serum in the AN group was significantly lower (p < 0.0001) than in the C group. The distribution of genotypes in the locus RETN c.62 of the RES gene was similar in both groups. A significant difference was demonstrated in the distribution of genotypes in the polymorphic site RETN c.-180 of the RES gene between AN and C (p = 0.0145) and in the distribution of the C and G alleles in the locus RETN c.-180 (p < 0.0001). The C allele occurred significantly more frequently than the G allele in the C group as compared to the AN group. In all the study subjects jointly (AN and C) a significant positive correlation between the blood RES levels on one hand and the body mass (r = 0.42; p < 0.0001) and BMI (r = 0.61; p< 0.0001) on the other was observed. There was no correlation between the concentration of RES in blood serum and the distribution of genotypes in the loci of the resistin gene referred to above. Conclusions: The CG genotype in the locus RETN c.-180 C>G of the RES gene may constitute one of the factors predisposing to the development of AN in girls. The genotype in the loci RETN c.62 G>A and RETN c.-180 C>G of the resistin gene has no influence on the levels of this hormone in blood in AN patients