Page 223 - 2025中醫藥與天然藥物聯合學術研討會-中醫藥與天然藥物的挑戰Ｘ機遇與未來大會手冊

PP-41

Geniposide protects against ferroptosis via ROS/p38 MAPK/NCOA4/GPX4-

mediated ferritinophagy in a co-culture system of HMC3 and SH-SY5Y cells

1
,1
Liang-Jun Wang, Y u -Chin Chang, Bin-Nan Wu*
1

1 Department of Pharmacology, Graduate Institute of Medicine, College of Medicine,
Kaohsiung Medical University, Kaohsiung, Taiwan
* E-mail: binnan@kmu.edu.tw

Abstract
Ferritinophagy is an important part of ferroptosis that regulates cell death related to reactive
oxygen species (ROS) and iron metabolism. Geniposide (GEN) is an iridoid glycoside and has
been reported for antioxidant and anti-inflammatory effects. Whether GEN can modulate
hydrogen peroxide (H2O2)-induced inflammation and ferroptosis and inflammation in human
neuroblastoma cells remains elusive. In the present study, we use the transwell co-culture
system, containing the human microglial cell line HMC3 (upper chamber) and SH-SY5Y cell
line SH-SY5Y (lower chamber), to investigate the mechanisms of action of GEN and its
neuroprotective effect. Our data indicate that GEN significantly inhibited ROS generation,
decreased the malondialdehyde (MDA) level, attenuated p38 MAPK phosphorylation, nuclear
receptor coactivator 4 (NCOA4), and SQSTM1 (p62) protein expression following H2O2
exposure. Meanwhile, GEN increases the intracellular GSH level and glutathione peroxidase 4
(GPX4) protein expression, protecting cells from oxidative damage. Therefore, GEN could
modulate NCOA4-mediated ferritinophagic flux and alleviate ferroptosis. In addition,
overexpression of NCOA4 significantly induced ferroptosis in both HMC3 and SH-SY5Y cells,
but GEN mitigated these effects. NCOA4-mediated ferritinophagy collectively contributes to
the process by which hydrogen peroxide induces oxidative stress and leads to cell ferroptosis.
In conclusion, GEN alleviates hydrogen peroxide-induced inflammation and ferroptosis
through inhibiting inflammatory responses and ferroptosis via the ROS/p38
MAPK/NCOA4/GPX4-mediated ferritinophagy signalling pathway.

Keywords: Co-culture system; Ferroptosis; Geniposide; Hydrogen peroxide; Oxidative stress