Presentation Slides - Institute for the Social Sciences

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.