Chronic liver disease (CLD) in children constitutes a major health burden on both parents and the diseased child. They may be caused by infectious, autoimmune, metabolic, vascular, drugs and toxins or unidentified etiologies. Many of these CLDs progress towards cirrhosis and eventually liver failure. In spite some of these disease categories are subjected to specific treatment with good prognosis, much are not, specially when there is no an identifiable etiology. Liver regulates the metabolic pathways and transport of trace elements, and consequently their bioavailability, tissue distribution and eventual toxicity. The liver also has a role in the excretion of trace elements through bile formation. Growing evidence indicates that many trace elements play important roles in a number of biological processes through activation or inhibition of enzymatic reactions, competing with other elements and metalloproteins for binding sites, and affecting the permeability of cell membranes. Moreover, some trace elements such as zinc (Zn), iron (Fe), and copper (Cu) exert important protective or enhancing effects on the progression of some diseases . Zinc is involved in stabilizing the cell membrane and prevents oxidative destruction caused by free radicals. The antioxidant actions of Zn include the induction of metallothionein (Zn-binding protein, formed by the liver), which is a potent scavenger of toxic metals and hydroxyl radical ). Fe and Cu ions catalyze the production of hydroxyl radical from hydrogen peroxide (H2O2). Zn is known to compete with both Fe and Cu for binding to cell membrane, thus decreasing the production of hydroxyl radical. Thus, it is clear that Zn has multiple roles as an antioxidant and is therefore, an excellent candidate for clinical chemoprevention trials in humans.

Serum copper, iron and zinc in children with chronic liver disease Noha L. Ibrahim*, Ahmad M. Sira**

* Molecular Diagnostics Departments, Genetic Engineering and Biotechnology Research Institute (GEBRI), Menofiya University, Egypt.

** Pediatrics Hepatology Department, National Liver Institute, Menofiya University, Egypt. INTRODUCTION

AIM OF THE WORKMeasurement of serum level of essential trace elements in children with CLD regardless the etiology and correlate these serum levels with biochemical measures of liver damage and other liver function tests.

CONCLUSIONIn conclusion, our finding of significantly higher serum Fe, Cu, and ferritin in children with CLD than healthy controls and its positive correlation with biochemical parameters of liver damage and the significantly lower serum Zn in CLD than that in healthy controls and its negative correlation with biochemical parameters of liver damage encourage the inclusion of these trace elements as biomarkers for monitoring the severity of liver damage during routine assessment of children with CLD. Finally, we recommend caution to avoid excess Fe and Cu intake in such patients, while zinc supplementation is encouraged regardless the etiology of CLD.

PATIENTS & METHODSStudy population

This study included 50 children with CLD (27 males and 23 females, with mean of age 5.86 ± 4.76 years). They were taken from the attendants of the outpatient and inpatient clinic of pediatric hepatology department, National Liver Institute, Menofiya University from October 2010 to February 2012. Another group of 50 age and sex matched healthy children were enrolled as a control group (25 males and 25 females, with mean of age 6.62 ± 3.54 years). None of the participants had received mineral supplements before blood sampling.

Etiological diagnosis

After full history taking, thorough clinical examination, the etiological diagnoses of the CLD group were autoimmune hepatitis (n=7), chronic hepatitis C (n=5), Alagille syndrome (n=5), cytomegalovirus hepatitis (n=4), progressive familial intrahepatic cholestasis (n=3), neglected biliary atresia (n=3), chronic hepatitis B (n=2), glycogen storage disease (n=2), Crigler Najjar syndrome (n=2), congenital hepatic fibrosis (n=2), Niemann Pick disease (n=2), inspissated bile syndrome (n=2), toxoplasma hepatitis (n=2), Caroli syndrome (n=2), portal vein thrombosis (n=2), Wilson disease (n=1), Dubin-Johnson syndrome (n=1), Budd Chiari syndrome (n=1), alpha1 antitrypsin deficiency (n=1) and cystic fibrosis (n=1).

Laboratory investigations

Liver function tests (Alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), gamma glutamyle transpeptidase (GGT), total bilirubin, direct bilirubin, total proteins and serum albumin) were measured using Automated Beckman Coulter. Serum Zn, Cu, and Fe were measured by colorimetric assay at a wave length of 560 nm (for Zn), 580 nm (for Cu), 546 nm (for Fe) using Biosystems BTS-310 photometer chemistry analyzer. TIBC was measured based on ferrozine method by Spectrum Diagnostic for the in-vitro quantitative, diagnostic determination of TIBC in human serum, serum ferritin was measured by ELISA (enzyme-linked immunosorbent assay) using the UBI MAGIWEL™ FERRITIN QUANTITATIVE device and serum transferrin saturation (TS) was calculated by dividing serum Fe by TIBC.

Statistical analysis

Descriptive results were expressed as mean ± SD (standard deviation) or number and percentage. For qualitative data, significance was tested using Chi square test. For quantitative data, significance between groups was tested using the Student's t- test for parametric data and by Mann-Whitney test for non-parametric data. Correlation was tested using Pearson’s correlation. Results were considered significant when P-value was less than 0.05. Statistical analysis was carried out using SPSS (statistical package for social science) program version 13 (SPSS Inc., Chicago, Illinois, USA) on an IBM compatible computer.

RESULTS

Box-and-whiskers plot for serum trace elements in the studied groups. The top and bottom of each box are the 75th and 25th centiles. The line through the box is the median and the error bars are the maximum and minimum. The horizontal bar represents the significance between the designated groups. Serum Cu, Fe, ferritin, and TS were significantly higher in the CLD than that in the control group; while Zn and TIBC were significantly lower in the CLD than that in the control group (P < 0.01)

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Figure 1: Comparison between the CLD and control group regarding age.

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Figure 2: Comparison between the CLD and control group regarding sex.

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Figure 3: Comparison of total protein, albumin, total and direct bilirubin between CLD

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Figure 4: Comparison of ALT,AST,ALP and GGT between CLD group and control group.

INTRODUCTION

AIM OF THE WORK

CONCLUSION

RESULTS

PATIENTS & METHODS