Assessing the efficacy of tocotrienol-rich fraction vitamin E in obese children with non-alcoholic fatty liver disease: a single-blind, randomized clinical trial

Abstract

Background: Childhood obesity is a growing concern, and non-alcoholic fatty liver disease (NAFLD) is a significant consequence. Currently, there are no approved drugs to treat NAFLD in children. However, a recent study explored the potential of vitamin E enriched with tocotrienol (TRF) as a powerful antioxidant for NAFLD. The aims of the present study were to investigate the effectiveness and safety of TRF in managing children with obesity and NAFLD.

Methods: A total of 29 patients aged 10 to 18 received a daily oral dose of 50 mg TRF for six months (January 2020 to February 2022), and all had fatty liver disease were detected by ultrasonography and abnormally high alanine transaminase levels (at least two-fold higher than the upper limits for their respective genders). Various parameters, including biochemical markers, FibroScan, LiverFASt, DNA damage, and cytokine expression, were monitored.

Results: APO-A1 and AST levels decreased significantly from 1.39 ± 0.3 to 1.22 ± 0.2 g/L (P = 0.002) and from 30 ± 12 to 22 ± 10 g/L (P = 0.038), respectively, in the TRF group post-intervention. Hepatic steatosis was significantly reduced in the placebo group from 309.38 ± 53.60 db/m to 277.62 ± 39.55 db/m (p = 0.048), but not in the TRF group. Comet assay analysis showed a significant reduction in the DNA damage parameters in the TRF group in the post-intervention period compared to the baseline, with tail length decreasing from 28.34 ± 10.9 to 21.69 ± 9.84; (p = 0.049) and with tail DNA (%) decreasing from 54.13 ± 22.1to 46.23 ± 17.9; (p = 0.043). Pro-inflammatory cytokine expression levels were significantly lower in the TRF group compared to baseline levels for IL-6 (2.10 6.3 to 0.7 1.0 pg/mL; p = 0.047 pg/mL) and TNF-1 (1.73 5.5 pg/mL to 0.7 0.5 pg/mL; p = 0.045).

Conclusion: The study provides evidence that TRF supplementation may offer a risk-free treatment option for children with obesity and NAFLD. The antioxidant and anti-inflammatory properties of TRF offer a promising adjuvant therapy for NAFLD treatment. In combination with lifestyle modifications such as exercise and calorie restriction, TRF could play an essential role in the prevention of NAFLD in the future. However, further studies are needed to explore the long-term effects of TRF supplementation on NAFLD in children.

Trial registration: The study has been registered with the International Clinical Trial Registry under reference number (NCT05905185) retrospective registration on (15/06/2023).

Keywords: Anti-inflammatory agent; Antioxidants; Non-alcoholic fatty liver disease; Oxidative stress; Tocotrienols.

Fig. 1

CONSORT flow diagram depicts how participants progressed through each stage of the randomized controlled trial, which used either tocotrienol-rich fraction vitamin E (TRF) or placebo

Fig. 2

Alkaline single-cell gel electrophoresis was used to assess DNA damage. A Baseline TRF-treated group cells with DNA damage visible as comets. B Baseline placebo-treated group cells with DNA damage shown as comets. After six months, cells with TRF supplementation had less DNA damage. Cells with increased DNA damage following six months of placebo supplementation

Fig. 3

DNA damage was determined using computerized image analysis for TRF and placebo at baseline and post-intervention. The comet assay method detected DNA damage in a whole blood sample by calculating the tail length (TL) and percentage of DNA in the tail (DNA%) of 100 randomly selected cells. A fluorescence microscope was used to examine the slide. ImageJ software was used to capture the photos and measure them. Data are presented as mean ± SEM, and a statistically significant value is deemed to be P < (0.05)

Fig. 4

Relative fold change of gene expression measured by quantitative real-time PCR for the vitamin E group & Placebo group at baseline and six months’ post-intervention. A Relative expression of IL-6 gene. B Relative expression of TNF-α. Relative expression of IFN-γ gene. Relative expression levels were normalised to GAPDH, and fold changes were calculated against the baseline. Values represent the means and SEM. Significant differences are highlighted as ∗ (P < 0.05)