Theaphenon E prevents fatty liver disease and increases CD4+ T cell survival in mice fed a high-fat diet - PubMed (original) (raw)

. 2021 Jan;40(1):110-119.

doi: 10.1016/j.clnu.2020.04.033. Epub 2020 May 4.

Ning Ma 2, Yanqi Hou 3, Eva Permaul 4, Deborah L Berry 5, M Idalia Cruz 6, Evan Pannkuk 7, Michael Girgis 8, Zizhao Zhu 9, Yichen Lee 10, Olga Rodriquez 11, Amrita Cheema 12, Fung-Lung Chung 13

Affiliations

Theaphenon E prevents fatty liver disease and increases CD4+ T cell survival in mice fed a high-fat diet

Heidi Coia et al. Clin Nutr. 2021 Jan.

Abstract

Background & aims: Obesity is a major cause of non-alcoholic fatty liver disease (NAFLD). NAFLD is an epidemic affecting nearly 34% of the adult population in the US. As a chronic inflammatory disease, NAFLD influences the immune system by dysregulating T-cell activity. Remedies for the adverse effects on the immune system are urgently needed. We studied Theaphenon E (TE), a standardized formulation of green tea extract, on the adverse effects of NAFLD in C57BL/6J mice fed a high fat diet (HFD).

Methods: Mice received HFD, low fat diet (LFD) or HFD+2% TE for 35 weeks. Hepatic lipid accumulation, cell proliferation, apoptosis and CD4+T lymphocytes were measured throughout the bioassay. The hepatic composition of fatty acids was determined. The effects of epigallocatechin gallate (EGCG) metabolites on lipid accumulation in mouse and primary human liver cells were studied.

Results: Unlike mice receiving HFD, mice on HFD+2% TE maintained normal liver to body weight ratios with low levels of alanine and aspartate aminotransferase (ALT and AST). Hepatic lipid accumulation was observed in HFD mice, accompanied by increased proliferation, reduced apoptosis and loss of CD4+ T lymphocytes. TE significantly inhibited lipid accumulation, decreased proliferation, induced apoptosis and increased CD4+ T cell survival in HFD mice. It was found that the EGCG metabolite EGC-M3 reduced lipid accumulation in mouse and human hepatocytes. Linoleic acid showed the largest increase (2.5-fold) in livers of mice on a HFD and this increase was significantly suppressed by TE.

Conclusions: Livers of HFD-fed mice showed lipid accumulation, increased proliferation, reduced apoptosis, elevated linoleic acid and loss of CD4+ T cells. TE effectively ameliorated all of these adverse effects.

Keywords: CD4+T cells; Linoleic acid; Non-alcoholic fatty liver disease; Prevention by green tea.

Published by Elsevier Ltd.

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Conflict of interest statement

Conflict of Interest The authors listed have no affiliations with or involvement in any organization or entity with any financial interest or non-financial interest in the subject matter or materials discussed in this manuscript.

Figures

Fig. 1.

Fig. 1.

Body weight curves of C57BL/6J mice over a period of 35 weeks of the experiment.

Fig. 2.

Fig. 2.

Liver size and weight. (A) The percent of the liver weight to body weight ratio of the mice collected at each time point and (B) Gross morphology of mice livers collected after 35 weeks on the diet. * indicates p < 0.05.

Fig. 3.

Fig. 3.

H&E staining of mouse liver tissue sections showing lipid accumulation in the liver from 0 to 35 weeks on each diet. Scale bar indicates 100 μm.

Fig. 4.

Fig. 4.

MRI images of fat content.

Fig. 5.

Fig. 5.

Serum levels of (A) ALT and (B) AST in the C57BL/6J mice on HFD, LFD and HFD + TE diets (*indicates p < 0.05).

Fig. 6.

Fig. 6.

Immunohistochemistry of HFD, LFD and HF + TE mouse liver tissues at 0 and 35 weeks and the percentages of positive cells across all time points (20X, scale bar indicates 200 μm, * indicates p < 0.05). (A) Ki67, (B) TUNEL and (C) CD4+.

Fig. 7.

Fig. 7.

(A) Nile Red staining of AML12 and (B) Primary human hepatocytes treated with fatty acids (FA) and Theaphenon E (TE) or no treatment (NT) after 1 and 3 days (D1 and D3). (20X).

Fig. 8.

Fig. 8.

(A) Nile Red staining of AML12 (20X) and (B) Primary human hepatocytes treated with fatty acids (FA) and EGCG metabolites EGC-M3, -M4 and -M5 after one and three days. (C) Structures of EGC-M3, EGC-M4 and EGC-M5.

Fig. 9.

Fig. 9.

Hepatic fatty acid changes in fold in HFD and HFD + TE fed mice compared to LFD at 35 weeks on diet. * indicates the total amount of 13 fatty acids that were measured.

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