Development of a study:
Role of inflammatory response
and oxidative stress in of fatty
liver disease in obese children
Richard Rosencrantz, Aliza
Solomon, David Blanco Hong Lin,
and Susanna Cunningham-Rundles
background
 Non alcoholic fatty liver disease is
increasing in children:
10-12% of normal weight children 2-19
years of age
 38% of obese children
 Obesity is associated with increased
inflammatory immune response and
increased leukocytes
Specific aims

Aim 1: Determine the significance of dysregulated
inflammatory cytokine response, altered immune cell
subsets, and oxidative stress for staging of pediatric
patients within the clinical spectrum of NAFLD
 Steatosis
 Steatohepatitis

Approach: Correlate biological markers of inflammation
such as elevated cytokines and urinary F2 isoprostanes
with liver function and non invasive detection of fat in liver
 Ultrasound
 MRI
Specific aims

Aim 2: Determine the effects of short term dietary
intervention on normalization of dysregulated inflammatory
cytokine response, altered immune cell subsets, and
oxidative stress in NAFLD
 Steatosis
 Steatohepatitis

Approach: Correlate biological markers of inflammation
such as elevated cytokines and urinary F2 isoprostanes
with liver function and non invasive detection of fat in liver
after one month supplementation with omega- 3 fatty acids
to isocaloric diet.
Pilot study
 16 obese patients (BMI>95%ile, mean age 11.5
years, range 3-17 years, 5 females
 Tests:




Standard liver function tests,
Creatinine-adjusted urinary F2-isoprostane
Lymphocyte immunophenotype subsets
Liver sonogram
 Comparisons:
 Steatohepatitis
 Hepatic steatosis group
 Obesity only
Problems with study
 Obesity based on BMI, no body
composition
 Limited diet history
 Limited discussion of diet and exercise
 Treatment recommendation limited to
Vitamin E
 Lack of followup
Liver enzymes
ALT Levels
100
90
80
70
60
50
40
30
20
10
0
OB
ST
SH
Limitation: comparatively few obese only controls
Lymphocyte subset differences

CD3+ T lymphocyte percentages reduced in SH
compared to age-matched controls (p=0.036)

CD3+/CD4+ T helper lymphocyte percentages
reduced in SH compared with age-matched
controls (p=0.01) and adult controls( p=0.03)

CD19+ B lymphocytes reduced in obese children
compared to age matched controls (p=0.03)

Limitation: historical age matched controls
Expression of CD95 among groups
Percent co expressing
CD3+CD95+ T Lymphocytes
60
50
40
30
20
10
0
SH
NFLD
OB
AC
 CD3+/CD95+
lymphocyte percentage
decreased in NAFLD
(p=0.015) and SH
groups (p=0.039)
compared to adult
controls
Limitation: need to follow up longitudinally
Urinary isoprostane in relationship to ALT
ALT vs. F2 Isoprostane
90.0
80.0
70.0
SH group
60.0
ALT
ST group
50.0
OB group
40.0
30.0
20.0
10.0
0.0
0
1,000
2,000
3,000
4,000
5,000
6,000
8-epi-prost ng/g creatinine
Limitation: few subjects and few obese only controls
Conclusions
 Direct measurements of oxidative stress
might be able to distinguish steatohepatitis
from steatosis in obese children.
 Lymphocyte subsets show distinct alterations
in obese children with and without NAFLD
but longitudinal studies are required to
determine significance.
 Clinical NAFLD in the pediatric population
appears to correlate with non-invasive
markers of oxidative stress.