Abstract
This research aimed to assess the impact of an aqueous extract from the cocoa shell and its major phytochemicals on preventing oxidative stress and mitochondrial dysfunction in hepatocytes using an in vitro model of nonalcoholic fatty liver disease (NAFLD). The phytochemicals from cocoa shell were extracted using water and characterized by UPLC-MS/MS analysis. HepG2 cells were cotreated with either the aqueous extract from cocoa shell (CAE, 20–100 µg mL−1) or 10–50 µmol L−1 of pure theobromine, protocatechuic acid, procyanidin B2, epicatechin, and catechin in the presence or absence of palmitic acid (PA, 500 µmol L−1) to mimic NAFLD conditions in vitro. Biomarkers of mitochondrial function and oxidative stress were evaluated 24 h after the cotreatment in cell supernatants and lysates using chemical, biochemical, and immunochemical techniques. CAE and the phytochemicals therein significantly (p < 0.05) protected mitochondrial content (15–100%) and preserved mitochondrial function, promoting O2 consumption (1.2- to 1.8-fold) and ATP production (1.3- to 2.1-fold). Phytochemicals from cocoa shell significantly (p < 0.05) decreased PA-triggered oxidative stress. The mitochondrial membrane potential was maintained (62–100%), and the production of mitochondrial superoxide (26–100%) and total ROS (17–100%) was abrogated. CAE significantly (p < 0.05) modulated cell signaling pathways associated with ROS production and mitochondrial dysfunction, including an increase in the phosphorylation of ERK1/2 (2.8-fold), protein kinase B (AKT) (2.8-fold), GSK3 (2.3-fold), Raf-1 (1.9-fold), and mTOR (1.7-fold). In conclusion, results suggested that the cocoa shell’s phytochemicals could protect mitochondrial liver function and alleviate oxidative stress by modulating key pathways involved in their regulation.
Highlights
In our previous studies [7,8], we demonstrated the applicability of phytochemicals extracted from cocoa shell in the modulation of inflammation, insulin resistance, and mitochondrial function in adipocytes
Cocoa Shell Was Mainly Composed of Theobromine, Protocatechuic Acid, and Flavan-3-Ols
We present the impact of a well-characterized theobromine- and phenolic-rich extract from cocoa shell and five pure bioactive compounds, assessing them in an in vitro model of nonalcoholic fatty liver disease (NAFLD)
Summary
The World Health Organization pinpoints chronic diseases as responsible for 71% of all deaths worldwide [1]. Metabolic disorders are caused by defective cellular metabolic processes triggering energy and redox imbalances and the induction of many pathophysiological conditions. The main hallmarks of metabolic disorders include obesity, insulin resistance, type 2 diabetes mellitus, hypertension, hyperlipidemia, and metabolic syndrome [2]. These conditions involve the development of nonalcoholic fatty liver disease (NAFLD), a disorder defined by excessive fat accumulation in the form of triglycerides (steatosis) in the liver [3]. The global prevalence of NAFLD is currently estimated to be
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