TY - JOUR
T1 - Synergistic effects of heterophyllin B with nintedanib against experimental pulmonary fibrosis in mice
AU - Chen, Ce
AU - Han, Zeyu
AU - Luo, Jiawen
AU - Wang, Jialin
AU - Liu, Tingting
AU - Zhang, Jie
AU - Zhang, Chaofeng
N1 - Publisher Copyright:
© 2025 Elsevier GmbH
PY - 2025/8
Y1 - 2025/8
N2 - Background: Pulmonary fibrosis (PF) is a chronic, lethal lung disease marked by permanent alterations to the lung tissue architecture. Although nintedanib (NDN) has been approved for clinical use, its therapeutic potential is substantially hampered by severe gastrointestinal side effects, notably diarrhea, which compromise patient adherence and quality of life. Purpose: This study aimed to investigate whether heterophyllin B (HB) could augment the antifibrotic efficacy of NDN while mitigating its gastrointestinal toxicity. Methods: The therapeutic potential of HB was evaluated in a bleomycin (BLM)-induced murine model of PF. Alterations in gut microbiota and serum metabolic profiles were determined via 16S rRNA gene sequencing and untargeted metabolomics, respectively. Mechanistic insights were performed in NCM460 colonic epithelial cells through IDO1 silencing, ferroptosis inhibition, CETSA and molecular dynamics experiments. Furthermore, the synergistic and protective effects of HB on NDN were investigated in BLM-induced mice, along with analysis of intestinal microbiota composition. The active constituents of the EtOAc extract of Radix Pseudostellariae were identified using UPLC-Q-TOF-MS/MS, GNPS, and NMR spectroscopy. Results: Administration of HB (40 mg/kg/day for approximately 14 days) significantly attenuated lung fibrosis progression and substantially alleviated diarrhea in BLM-induced PF mice. HB reshaped the intestinal microecology and reprogrammed serum metabolism, notably by reducing the abundance of Escherichia-Shigella, as revealed by 16S rRNA sequencing and untargeted metabolomics analyses. Furthermore, the co-treatment of HB and NDN demonstrated enhanced efficacy and reduced gastrointestinal toxicity both in vivo and in vitro. Mechanistic investigations indicated that HB-enriched 3-hydroxybutyric acid (3-HA) restored intestinal mucosal barrier integrity by inhibiting IDO1-mediated ferroptosis. Additionally, extracts of Radix Pseudostellariae containing HB-like cyclopeptides significantly improved PF symptoms and intestinal epithelial injury in BLM-induced mice. Nine cyclopeptide compounds (Herterophyllin A-B, D and Psedostellarin A-E, G) were identified in the extract via UPLC-Q-TOF-MS/MS analysis. Conclusion: HB offers dual protection against pulmonary fibrosis and intestinal damage through its regulatory impact on the gut–lung axis and suppression of ferroptosis mechanisms. Collectively, HB offers a promising adjuvant to optimize NDN-based antifibrotic therapy, offering a novel strategy for integrated pulmonary and gastrointestinal protection in PF management.
AB - Background: Pulmonary fibrosis (PF) is a chronic, lethal lung disease marked by permanent alterations to the lung tissue architecture. Although nintedanib (NDN) has been approved for clinical use, its therapeutic potential is substantially hampered by severe gastrointestinal side effects, notably diarrhea, which compromise patient adherence and quality of life. Purpose: This study aimed to investigate whether heterophyllin B (HB) could augment the antifibrotic efficacy of NDN while mitigating its gastrointestinal toxicity. Methods: The therapeutic potential of HB was evaluated in a bleomycin (BLM)-induced murine model of PF. Alterations in gut microbiota and serum metabolic profiles were determined via 16S rRNA gene sequencing and untargeted metabolomics, respectively. Mechanistic insights were performed in NCM460 colonic epithelial cells through IDO1 silencing, ferroptosis inhibition, CETSA and molecular dynamics experiments. Furthermore, the synergistic and protective effects of HB on NDN were investigated in BLM-induced mice, along with analysis of intestinal microbiota composition. The active constituents of the EtOAc extract of Radix Pseudostellariae were identified using UPLC-Q-TOF-MS/MS, GNPS, and NMR spectroscopy. Results: Administration of HB (40 mg/kg/day for approximately 14 days) significantly attenuated lung fibrosis progression and substantially alleviated diarrhea in BLM-induced PF mice. HB reshaped the intestinal microecology and reprogrammed serum metabolism, notably by reducing the abundance of Escherichia-Shigella, as revealed by 16S rRNA sequencing and untargeted metabolomics analyses. Furthermore, the co-treatment of HB and NDN demonstrated enhanced efficacy and reduced gastrointestinal toxicity both in vivo and in vitro. Mechanistic investigations indicated that HB-enriched 3-hydroxybutyric acid (3-HA) restored intestinal mucosal barrier integrity by inhibiting IDO1-mediated ferroptosis. Additionally, extracts of Radix Pseudostellariae containing HB-like cyclopeptides significantly improved PF symptoms and intestinal epithelial injury in BLM-induced mice. Nine cyclopeptide compounds (Herterophyllin A-B, D and Psedostellarin A-E, G) were identified in the extract via UPLC-Q-TOF-MS/MS analysis. Conclusion: HB offers dual protection against pulmonary fibrosis and intestinal damage through its regulatory impact on the gut–lung axis and suppression of ferroptosis mechanisms. Collectively, HB offers a promising adjuvant to optimize NDN-based antifibrotic therapy, offering a novel strategy for integrated pulmonary and gastrointestinal protection in PF management.
KW - Ferroptosis
KW - Heterophyllin B
KW - Intestinal microbiota
KW - Nintedanib
KW - Pulmonary fibrosis
UR - http://www.scopus.com/inward/record.url?scp=105007644481&partnerID=8YFLogxK
U2 - 10.1016/j.phymed.2025.156922
DO - 10.1016/j.phymed.2025.156922
M3 - Article
AN - SCOPUS:105007644481
SN - 0944-7113
VL - 144
JO - Phytomedicine
JF - Phytomedicine
M1 - 156922
ER -
