Urinary and serum oxysterols in children: developmental pattern and potential biomarker for pediatric liver disease

Abstract

few reports describe oxysterols in healthy children or in children with liver disease. We aimed to determine whether developmental changes in urinary and serum oxysterols occur during childhood, and to assess whether oxysterols might be biomarkers for pediatric liver disease. Healthy children enrolled as subjects (36 and 35 for urine and serum analysis, respectively) included neonates, infants, preschoolers, and school-age children, studied along with 14 healthy adults and 8 children with liver disease. We quantitated 7 oxysterols including 4β-, 20(S)-, 22(S)-, 22(R)-, 24(S)-, 25-, and 27-hydroxycholesterol using liquid chromatography/electrospray ionization-tandem mass spectrometry. Urinary total oxysterols were significantly greater in neonates than in infants (P < 0.05), preschoolers (P < 0.001), school-age children (P < 0.001), or adults (P < 0.001), declining with age. Serum total oxysterols in neonates were significantly lower than in infants (P < 0.05), preschoolers (P < 0.001), school-age children (P < 0.05), or adults (P < 0.01). Compared with healthy children, total oxysterols and 24(S)-hydroxycholesterol in liver disease were significantly increased in both urine (P < 0.001 and P < 0.001, respectively) and serum (P < 0.001 and P < 0.05, respectively). Oxysterols in liver disease, particularly 24(S)-hydroxycholesterol, were greater in urine than serum. Oxysterols change developmentally and might serve as a biomarker for pediatric liver disease. to our knowledge, this is the first such report.

A Japanese prospective multicenter study of urinary oxysterols in biliary atresia

“…Non-BA diagnoses included NICCD in 3 patients, Alagille syndrome in 2, unknown in 2, neonatal leukemia in 1, Dubin-Johnson syndrome in 1, and hypothyroidism in 1. Two subjects with BA and 3 with non-BA were also enrolled in our previous study 21 , as were 3 HC. Characteristics of BA and non-BA patients are shown in Table 1 .…”

“…This data supports the view that 27-hydroxycholesterol is increased in BA because of acidic pathway upregulation. Unfortunately, we could not evaluate oxysterols in the classical pathway such as 7α-hydroxycholesterol, 7β-hydroxycholesterol, and 7-oxo-cholesterol because of previously reported oxidation effects 21 , 31 . 7α-Hydroxy-4-choesten-3-one, which reflects the condition of classical pathway, also was not examined 32 .…”

“…We quantitatively analyzed 7 types of oxysterols including 4β-hydroxycholesterol (cholest-5-en-3β,4β-diol), 20(S)-hydroxycholesterol (cholest-5-en-3β,20S-diol), 22(S)-hydroxycholesterol (cholest-5-en-3β,22S-diol), 22(R)-hydroxycholesterol (cholest-5-en-3β,22R-diol), 24(S)-hydroxycholesterol (cholest-5-en-3β,24S-diol), 25-hydroxycholesterol (cholest-5-en-3β,25-diol), and 27-hydroxycholesterol (cholest-5-en-3β,26-diol) using liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI–MS/MS) according to a previously reported method 21 , 31 . Urinary concentrations of individual oxysterols were corrected for creatinine concentration and are expressed as micromoles per moles of creatinine 21 , 31 .…”

“…Urine samples were prepared by enzymatic hydrolysis of oxysterol conjugates according to a reported method 20 , 21 , 33 , 34 . Forty microliters of reagent A and ten microliters of reagent B from the Glufatase set were added to 200 µL of sample.…”

“…So far we know of no prior studies considering oxysterols as potentially useful markers for BA. Recently, we reported that urinary oxysterols follow a developmental pattern in healthy children; deviations might suggest pediatric liver disease 21 . We hypothesized that urinary oxysterol determinations could help in distinguishing BA from other causes of neonatal cholestasis such as NICCD and Alagille syndrome, and examined the utility of oxysterol measurements in this prospective multicenter Japanese study.…”