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. 2025 May 20;20(5):e0324164.
doi: 10.1371/journal.pone.0324164. eCollection 2025.

Therapeutic potential of a choline-zinc-vitamin E nutraceutical complex in ameliorating thioacetamide-induced nonalcoholic fatty liver pathology in zebrafish

Bingbing Cao  1 , Jiali Zhou  1 , Bo Xia  1 , Xiaoqing Li  2 , Rui Wang  2 , Yiqiao Xu  1 , Chunqi Li  1
Affiliations

Therapeutic potential of a choline-zinc-vitamin E nutraceutical complex in ameliorating thioacetamide-induced nonalcoholic fatty liver pathology in zebrafish

Bingbing Cao et al. PLoS One. .

Abstract

Choline has been proven to be effective in maintaining liver function. However, the effect of choline, in combination with other nutrients, on the improvement of non-alcoholic fatty liver disease (NAFLD) remains unclear. This study aimed to investigate the potential effect of the nutraceutical complex containing choline bitartrate, zinc citrate, and dl-α-Tocopheryl acetate on NAFLD in the zebrafish model. The NAFLD model was induced in zebrafish by administering thioacetamide. Experimental groups were established, including a normal control group, the model control group, the positive control group, the nutraceutical complex intervention group, and the choline bitartrate alone intervention group. The intervention was administered to the zebrafish in a water-soluble form, while the positive control group received polyene phosphatidylcholine at a concentration of 50.0 μg/mL. Notably, the protective effect of the nutraceutical complex against NAFLD is more pronounced than that observed with choline bitartrate supplementation alone. The results of transcriptomics and quantitative real-time PCR showed that the potential mechanisms underlying the effects of the nutraceutical complex might involve the upregulation of acacia, acsl1a, fbp2 gene expression, and the downregulation of tbc1d1 gene expression. These results were further validated by western blotting and overexpression experiments. Our findings indicated that choline bitartrate, zinc citrate, and dl-α-Tocopheryl acetate can help improve NAFLD. The results of this study provide evidence for the application of the nutraceutical complex in the improvement of NAFLD.

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Conflict of interest statement

Xiaoqing Li and Rui Wang are currently employees of Opella. Bingbing Cao, Jiali Zhou, Bo Xia, Yiqiao Xu and Chunqi Li are employees of Hunter Biotechnology, Inc. which performed the study by order of Opella. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1
Fig 1. Intensity of zebrafish liver fat staining after treatment with nutraceutical complex.
(A) Normal control group; (B) Model control group; (C) The positive control group: polyunsaturated phosphatidylcholine 50.0 μg/mL; (D) Low dose of nutraceutical complex: 125 μg/mL; (E) Middle dose of nutraceutical complex: 250 μg/mL; (F) High dose of nutraceutical complex: 500 μg/mL; (G) Low dose of choline bitartrate control group: 81.0 μg/mL; (H) Middle dose of choline bitartrate control group: 162 μg/mL; (I) High dose of choline bitartrate control group: 316 μg/mL. The location of the liver is marked with a yellow dotted line in the example figure.
Fig 2
Fig 2. Liver tissue structure in zebrafish after treatment with nutraceutical complex.
(A) Normal control group; (B) Model control group; (C) The positive control group: polyunsaturated phosphatidylcholine 50.0 μg/mL; (D) Low dose of nutraceutical complex: 125 μg/mL; (E) Middle dose of nutraceutical complex: 250 μg/mL; (F) High dose of nutraceutical complex: 500 μg/mL; (G) Low dose of choline bitartrate control group: 81.0 μg/mL; (H) Middle dose of choline bitartrate control group: 162 μg/mL; (I) High dose of choline bitartrate control group: 316 μg/mL. The black dashed box indicates the observed area in the example picture, and the red arrow points to vacuolar degeneration in the model control group.
Fig 3
Fig 3. Fluorescence intensity graph of zebrafish liver fat after treatment with nutraceutical complex.
(A) Normal control group; (B) Model control group; (C) The positive control group: polyunsaturated phosphatidylcholine 50.0 μg/mL; (D) Low dose of nutraceutical complex: 125 μg/mL; (E) Middle dose of nutraceutical complex: 250 μg/mL; (F) High dose of nutraceutical complex: 500 μg/mL; (G) Low dose of choline bitartrate control group: 81.0 μg/mL; (H) Middle dose of choline bitartrate control group: 162 μg/mL; (I) High dose of choline bitartrate control group: 316 μg/mL. Yellow dashed line indicates the liver.
Fig 4
Fig 4. The protein expression of fbp2, acsl1, tbc1d1, and acaca.
Fig 5
Fig 5. Relative expression of acsl1a gene.
*Comparison with normal control group, p < 0.05.
Fig 6
Fig 6. Typical plot of zebrafish liver fat staining intensity after sample treatment.
The yellow dashed box is the liver.
See this image and copyright information in PMC

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