Truffle Extract Attenuates TGF-β1-Induced Fibrosis-Associated Responses in HFL-1 Cells and Is Associated with Changes in Smad and MAPK Signaling
6 affiliations
- 1Doctor Program in Biotechnology Industrial Management and Innovation, National Chung Hsing University, Taichung 40227, Taiwan
- 2Institute of Molecular Biology, National Chung Hsing University, Taichung 40227, Taiwan
- 3iEGG and Animal Biotechnology Center, National Chung Hsing University, Taichung 40227, Taiwan
- 4Division of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, Lancaster LA1 4YQ, UK
- 5Rong Hsing Research Center for Translational Medicine, National Chung Hsing University, Taichung 40227, Taiwan
- 6Department of Life Sciences, National Chung Hsing University, Taichung 40227, Taiwan
Abstract
Cellular fibrosis is a key pathological feature of chronic disease and is characterized by persistent fibroblast activation and excessive extracellular matrix deposition. In this study, an aqueous truffle extract (TE) was characterized by untargeted LC-ESI(+)-MS/MS and LC-ESI(−)-MS/MS profiling that yielded putative LC-MS/MS annotations, and was evaluated in a TGF-β1-induced HFL-1 fibroblast model. TE attenuated TGF-β1-induced Smad2 phosphorylation, Smad2/3-Smad4 association, nuclear accumulation of Smad2/3 and Smad4, and the expression of the fibrosis-associated proteins α-smooth muscle actin (α-SMA) and fibronectin. TGF-β1 increased MEK1/2 phosphorylation, whereas TE further increased p-MEK1/2 while decreasing Ras expression and p-ERK1/2, revealing a divergent, context-dependent MAPK response rather than uniform activation or inhibition of the Ras/MEK/ERK pathway. shRNA-mediated reduction in TGF-β1, MEK1/2, or ERK1/2 altered fibrosis-associated protein expression in the absence of TE and further modified the protein-expression patterns observed after TE treatment, indicating that the TE response occurred within a signaling context influenced by these components without establishing strict pathway dependence or a linear signaling hierarchy. In the DCFH-DA assay, TGF-β1 increased intracellular oxidation-dependent fluorescence, whereas both TGF-β1 plus TE and TGF-β1 plus N-acetyl-L-cysteine reduced the signal. LC-MS/MS profiling additionally provided exploratory compositional context for the tested aqueous preparation, while the database best matches and putative LC-MS/MS annotation names remain exploratory and unconfirmed. Collectively, these findings show that TE attenuates TGF-β1-induced fibrosis-associated responses in HFL-1 cells in association with suppression of canonical Smad signaling, reduced intracellular oxidative activity, and context-dependent remodeling of MAPK-network readouts. These coordinated changes suggest that the antifibrotic-associated activity of TE involves multiple interconnected cellular processes rather than modulation of a single signaling component. The results support further activity-guided chemical characterization and validation in additional fibroblast and in vivo fibrosis models.
Related
- Ellagic Acid Attenuates High-Fat, High-Sucrose Diet-Induced Cardiac Fibrosis by Suppressing AGE–RAGE Signaling, Oxidative Stress, and Inflammation
- Emerging Roles of FGFR3 and FGF23-Associated Signaling in Renal Fibrosis: Molecular Integration, Clinical Correlates, and Therapeutic Perspectives
- Fucoidan attenuates metabolic dysfunction-associated fatty liver disease in mice and is associated with gut microbiota remodeling and AMPK/PPARγ signaling
- LED Photobiomodulation at 630 nm Attenuates TNF-α-Induced Senescence-Associated Responses in Association with Mitochondrial Homeostasis
- Integrated Transcriptomic and Biochemical Analyses Reveal Coordinated Suppression of ER-Associated Survival Signaling and Activation of ROS-Mediated Stress Responses by Oxidized Graphene Quantum Dots in Estrogen Receptor-Positive Breast Cancer
BibTeX
@article{sun2026truffle,
title = {{Truffle Extract Attenuates TGF-β1-Induced Fibrosis-Associated Responses in HFL-1 Cells and Is Associated with Changes in Smad and MAPK Signaling}},
author = {Te-Kai Sun and Jyun-Yi Li and Yu-Wen Sun and Jing-Wen Huang and Yi-Ying Wu and Muhammad Munir and Hung-Jen Liu},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = nov,
doi = {10.3390/ijms272110030},
url = {https://www.mdpi.com/1422-0067/27/21/10030},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Truffle Extract Attenuates TGF-β1-Induced Fibrosis-Associated Responses in HFL-1 Cells and Is Associated with Changes in Smad and MAPK Signaling
AU - Te-Kai Sun
AU - Jyun-Yi Li
AU - Yu-Wen Sun
AU - Jing-Wen Huang
AU - Yi-Ying Wu
AU - Muhammad Munir
AU - Hung-Jen Liu
PY - 2026
DA - 2026/11/09
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms272110030
UR - https://www.mdpi.com/1422-0067/27/21/10030
AB - Cellular fibrosis is a key pathological feature of chronic disease and is characterized by persistent fibroblast activation and excessive extracellular matrix deposition. In this study, an aqueous truffle extract (TE) was characterized by untargeted LC-ESI(+)-MS/MS and LC-ESI(−)-MS/MS profiling that yielded putative LC-MS/MS annotations, and was evaluated in a TGF-β1-induced HFL-1 fibroblast model. TE attenuated TGF-β1-induced Smad2 phosphorylation, Smad2/3-Smad4 association, nuclear accumulation of Smad2/3 and Smad4, and the expression of the fibrosis-associated proteins α-smooth muscle actin (α-SMA) and fibronectin. TGF-β1 increased MEK1/2 phosphorylation, whereas TE further increased p-MEK1/2 while decreasing Ras expression and p-ERK1/2, revealing a divergent, context-dependent MAPK response rather than uniform activation or inhibition of the Ras/MEK/ERK pathway. shRNA-mediated reduction in TGF-β1, MEK1/2, or ERK1/2 altered fibrosis-associated protein expression in the absence of TE and further modified the protein-expression patterns observed after TE treatment, indicating that the TE response occurred within a signaling context influenced by these components without establishing strict pathway dependence or a linear signaling hierarchy. In the DCFH-DA assay, TGF-β1 increased intracellular oxidation-dependent fluorescence, whereas both TGF-β1 plus TE and TGF-β1 plus N-acetyl-L-cysteine reduced the signal. LC-MS/MS profiling additionally provided exploratory compositional context for the tested aqueous preparation, while the database best matches and putative LC-MS/MS annotation names remain exploratory and unconfirmed. Collectively, these findings show that TE attenuates TGF-β1-induced fibrosis-associated responses in HFL-1 cells in association with suppression of canonical Smad signaling, reduced intracellular oxidative activity, and context-dependent remodeling of MAPK-network readouts. These coordinated changes suggest that the antifibrotic-associated activity of TE involves multiple interconnected cellular processes rather than modulation of a single signaling component. The results support further activity-guided chemical characterization and validation in additional fibroblast and in vivo fibrosis models.
SN - 1422-0067
ER -