Randomised Clinical Trial: Calorie Restriction Regimen with Tomato Juice Supplementation Ameliorates Oxidative Stress and Preserves a Proper Immune Surveillance Modulating Mitochondrial Bioenergetics of T-Lymphocytes in Obese Children Affected by Non-Alcoholic Fatty Liver Disease (NAFLD)

Abstract

Fatty liver disease is a serious complication of childhood obesity. Calorie-restricted regimen (RCR) is one of the effective therapy for this condition. Aim of the study was to evaluate the effect of lycopene-rich tomato sauce with oregano and basil extracts in obese children with fatty liver on RCR. 61 obese children with fatty liver were enrolled, 52 completed the study. A randomized cross over clinical trial was performed. Participants were assigned to RCR alone or with a supplement of lycopene-rich tomato juice for 60 days; subsequently, the groups were switched to the alternative regimen for the next 60 days. Reduction in BMI, HOMA-IR, cholesterol, triglycerides, liver size, and steatosis was more profound in tomato-supplemented group. Leptin decreased in both groups whereas adiponectin raised only after tomato supplementation. RCR is associated with the impaired engagement of T-cells glycolysis and proliferation, tomato-supplementation resulted in glycolytic metabolic activation of T-cells. Tomato juice ameliorates glucose and lipid metabolism in obese children, improve oxidative and inflammatory state and modulates the mitochondrial metabolism of T-cells contributing to a maintenance of a proper immune surveillance in children, impaired by RCR. The addition of tomato to RCR could be considered a protective and preventive support to obese child.

Keywords: NAFLD; immunology; inflammation; nutrition; pediatric hepatology.

Figure 1

Percentage changes of anthropometric measures (A) and serum biochemical parameters (B) at 60 and 120 days compared to baseline (T0) (see study design). White = RCR; Red = RCR + tomato. Statistically significant differences are reported in Tables S3 and S4.

Figure 2

Changes in oxidative stress markers at 60 and 120 days compared to baseline. White = RCR, Red = RCR + tomato; Serum levels of (A) oxidized low-density lipoprotein (LDL); (B) carbonylated proteins (PC), (C) malondialdehyde (MDA); (D,E) reduced (GSH) and oxidized glutathione (GSSG) and (F) GSH to GSSG ratio. ** p < 0.01 paired Student’s-test comparing the effect of each intervention to the baseline. Oxidized LDL and carbonylated proteins were significantly reduced in both groups over tomato supplementation (Figure 2A,B). MDA decreased in both groups at T1 compared to T0 (Figure 2C) (−23% p = 0.000 for RCR + Tomato group, –15% p = 0.000 for RCR group), GSH increased and GSSG decreased in group 2 (RCR + Tomato), both at T1 and T2, compared to baseline values (Figure 2D,E). Consequently, GSH/GSSG ratio increased in group 2 at both time points (Figure 2F). In group 1 (RCR) GSH and GSH/GSSG ratio increased only at T2 (after RCR + Tomato) compared to baseline values, whereas GSSG did not change (Figure 2D–F).

Figure 3

Percentage changes in ultrasound parameters at 60 and 120 days compared to baseline. Statistically significant differences are reported in Table S6. (A) Body fat; (B) Liver size; (C) Hepato-Renal Index, HRI. White = RCR, Red = RCR + T.

Figure 4

Stratification of patients by degrees of steatosis based on ultrasound findings at the baseline, 60 and 120 days. (A) group 1; (B) group 2.

Figure 5

Percentage changes in serum IL-4 and adipokines levels at 60 and 120 days compared to baseline. White = RCR, Red = RCR+T. * p ≤ 0.05, ** p ≤ 0.01 Wilcoxon-test comparing the effect of each intervention to the baseline.

Figure 6

Percentage changes in metabolic parameters (A) Glycolysis; (B) Mitochondrial respiration) of PBMCs isolated from children at 60 and 120 days compared to baseline upon 12 h OKT3 stimulation. White = RCR, Red = RCR + Tomato. Statistically significant differences are reported in Figure S3.