Enterococcus faecalis
CGz3 alleviating steatosis via BSH-mediated modulation in HepG2 cell-lines
Koujalagi Koushik and Malaviya Alok
Res. J. Biotech.; Vol. 20(12); 92-100;
doi: https://doi.org/10.25303/2012rjbt0920100; (2025)
Abstract
The study aimed to evaluate the therapeutic potential of bile salt hydrolase (BSH)-
producing probiotic Enterococcus faecalis CGz3 in alleviating steatosis in HepG2
hepatocarcinoma cells, with non-alcoholic fatty liver disease (NAFLD) induced by
cholesterol and oleic acid (OA), focusing on its effects on lipid accumulation,
metabolic gene expression and, inflammatory pathways. HepG2 cells were treated with
cholesterol and OA to induce lipid accumulation, mimicking non-alcoholic fatty liver
disease (NAFLD) conditions. Cells were then incubated with E. faecalis CGz3 for
6 hours at 37°C and 5% CO₂. Lipid levels were quantified using Oil Red O staining
and cholesterol uptake assays, while gene expression of lipogenic, inflammatory
and metabolic markers was assessed via quantitative real-time polymerase chain reaction
(qRT-PCR).
Treatment with E. faecalis CGz3 significantly reduced lipid accumulation from 42.96±1.35
mg/mL in NAFLD-induced cells to 29.73±1.26 mg/mL. It down regulated lipogenic genes
(SREBP-1c, FAS and ACC) and inflammatory markers (TNF-α, IL-6, CRP, TLR4, TLR9,
NF-κB, JNK, ERK) while upregulating PPARα and AMPK, promoting fatty acid oxidation.
No significant cytotoxicity was observed at 6 hours, though prolonged exposure (12–24
hours) reduced cell viability. This study introduces E. faecalis CGz3, a novel BSH-producing
probiotic isolated from chicken gizzard, as a promising candidate for NAFLD intervention.
Its selective modulation of lipid metabolism and inflammation via BSH activity offers
a new perspective on probiotic-based therapies for NAFLD, warranting further in
vivo and clinical exploration.
