Differences in age at menarche according to BMI z-score tertiles.

<p>*p<0.001.</p

Clinical data of 240 obese Italian girls according to CB2 Q63R variant.

<p>SD: standard deviation; BMI: body mass index; W/ Ht r: waist/height ratio; <i>p</i>-values <0.05 have been considered significant and are shown in bold.</p><p>Clinical data of 240 obese Italian girls according to CB2 Q63R variant.</p

Cannabinoid Receptor <em>Type 2</em> Functional Variant Influences Liver Damage in Children with Non-Alcoholic Fatty Liver Disease

<div><p>Non-alcoholic fatty liver disease (NAFLD) comprises a spectrum of disease ranging from simple steatosis to inflammatory steatohepatitis (NASH) with different degrees of fibrosis that can ultimately progress to cirrhosis. Accumulating evidence suggests the involvement of the endocannabinoid-system in liver disease and related complications. In particular, hepatoprotective properties for Cannabinoid Receptor <em>type 2</em> (CB2) have been shown both through experimental murine models of liver injury and association study between a CB2 functional variant, Q63R, and liver enzymes in Italian obese children with steatosis.</p> <p>Here, in order to clarify the role of CB2 in severity of childhood NAFLD, we have investigated the association of the CB2 Q63R variant, with histological parameters of liver disease severity in 118 Italian children with histologically-proven NAFLD.</p> <p>CB2 Q63R genotype was assigned performing a TaqMan assay and a general linear model analysis was used to evaluate the association between the polymorphism and the histological parameters of liver damage.</p> <p>We have found that whereas CB2 Q63R variant is not associated with steatosis or fibrosis, it is associated with the severity of the inflammation (<em>p</em> = 0.002) and the presence of NASH (<em>p</em> = 0.02).</p> <p>Our findings suggest a critical role for CB2 Q63R variant in modulating hepatic inflammation state in obese children and in the consequent increased predisposition of these patients to liver damage.</p> </div

Clinical and laboratory characteristics of obese patients subdivided by PNPLA3 I148M genotype.

<p>Values are expressed as means ± standard deviations. GLM analysis including gender, age and pubertal stage as covariates has been used to compare continuous variables. Abbreviations: BMI-SDS: Body Mass Index Standard Deviation Scores; W/Hr: Waist circumference to height ratio; HOMA: homeostatic model of assessment of insulin resistance index; WBISI: whole body insulin sensitivity index; HDL-C: high density lipoprotein-cholesterol; ALT: alanine transaminase; AST: aspartate transaminase; Gamma-GT: Gamma-Glutamyl transferase.</p><p>*WBISI was available in 497 patients.</p

Risk of pathologic ALT levels in 1048 obese children stratified in six categories according to fat abdominal size (W/Hr) and PNPLA3 genotype.

<p>ALT (alanine transaminase) values are expressed as means ± standard deviations. Logistic regression analysis has been used to calculate the Odds Ratios (OR) to have ALT>40 UI/L for each category of patients (from I to VI), compared to the entire cohort of children.</p

<i>CNR2</i> rs35761398 genotype and susceptibility to NASH.

<p><b>A</b>) Relationship between the <i>CNR2</i> rs35761398 genotype and the presence of NASH in 118 children with NAFLD (<i>p</i> = 0.02). Fifty-three out of 118 patients show definitive NASH. Among these NASH subjects 2 were homozygous for the CB2 Q63 allele, 23 were CB2 Q63R heterozygous and 28 homozygous for the CB2 R63. <b>B</b>) Relationship between the <i>CNR2</i> rs35761398 genotype and the presence of NASH in 55 I148M PNPLA3 heterozygous children with NAFLD (<i>p</i> = 0.001). Among I148M PNPLA3 subjects 38 out of 55 show definitive NASH and were all QR or RR for the CB2 Q63R variant. Age, sex, waist circumference and HOMA-IR index have been used as covariates.</p

Association between ALT levels and W/Hr, HOMA and BMI-SDS according to the PNPLA3 genotype.

<p><b>A</b>: Regression analysis describing the relationship between ALT levels and W/Hr in patients homozygous for <i>PNPLA3</i> M variant, heterozygous, and homozygous for PNPLA3 I variant. The regression between ALT levels and W/Hr in the group of patients homozygous for <i>PNPLA3</i> M/M is described by the equation y = 4.6+2.4*× (r = 0.36; p = 0.00001). The equation for <i>PNPLA3</i> I/M was y = 3.7+1.4*× (r = 0.22; p = 0.00001). The equation for PNPLA3 I/I was y = 3.4+1.1*× (r = 0.17; p = 0.0005). The comparison between the three regression lines is significant (p = 0.0045). <b>B</b>: Regression analysis describing the relationship between ALT levels and HOMA-IR in patients homozygous for <i>PNPLA3</i> M variant, heterozygous, and homozygous for PNPLA3 I variant. The regression between ALT levels and HOMA-IR in the group of patients homozygous for <i>PNPLA3</i> M/M is described by the equation y = 3.2+0.14*× (r = 0.18; p = 0.02). The equation for <i>PNPLA3</i> I/M was y = 3.0+1.12*× (r = 0.16; p = 0.001). The equation for PNPLA3 I/I was y = 2.9+0.16*× (r = 0.23; p = 0.00005). The three equations are not significantly different as to slopes (p = 0.7). <b>C</b>: Regression analysis describing the relationship between ALT levels and BMI z-score in patients homozygous for <i>PNPLA3</i> M variant, heterozygous, and homozygous for PNPLA3 I variant. The regression between ALT levels and BMI z-score in the group of patients homozygous for <i>PNPLA3</i> M/M is described by the equation y = 3.1+0.24*× (r = 0.01; p = 0.13). The equation for <i>PNPLA3</i> I/M was y = 3.1+0.17*× (r = 0.08; p = 0.14). The equation for PNPLA3 I/I was y = 2.9+0.17*× (r = 0.09; p = 0.04). The three equations are not significantly different as to slopes (p = 0.5).</p

Clinical and Laboratory Characteristics of the 118 Italian Pediatric Patients with biopsy-proven NAFLD.

<p>Values are expressed as means ± standard deviations. Abbreviations: BMI: Body Mass Index; HOMA-IR: homeostatic model of assessment of insulin resistance; ISI: insulin sensitivity index; IGT: Impaired Glucose Tolerance; ALT: alanine transaminase; AST: aspartate transaminase; Gamma-GT: Gamma-Glutamyl transferase; NASH: nonalcoholic steatohepatitis; G = grading.</p

Clinical and laboratory characteristics of the 118 Italian children with NAFLD stratified by the rs35761398 <i>CNR2</i> SNP encoding for the Q63R CB2 protein variant.

<p>Values are expressed as means ± standard deviations or as numbers and percentages. Mean values are compared by analysis of ANOVA. GLM analysis including gender, age, waist circumference and HOMA-IR as covariates has been used to compare continuous variables. Abbreviations: BMI: Body Mass Index; HOMA-IR: homeostatic model of assessment of insulin resistance; ISI: insulin sensitivity index; IGT: Impaired Glucose Tolerance; ALT: alanine transaminase; AST: aspartate transaminase; Gamma-GT: Gamma-Glutamyl transferase.</p

Clinical and laboratory characteristics of the 1048 children involved in the study.

<p>Values are expressed as means ± standard deviations. Ranges are in brackets. Abbreviations: BMI-SDS: Body Mass Index Standard Deviation Scores; W/Hr: Waist circumference to height ratio; HOMA-IR: homeostatic model of assessment of insulin resistance index; WBISI: whole body insulin sensitivity index; HDL-C: high density lipoprotein-cholesterol; ALT: alanine transaminase; AST: aspartate transaminase; Gamma-GT: Gamma-Glutamyl transferase. * WBISI was available in 497 patients.</p