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Mechanism of dietary iron overload inducing hepatocyte damage in
grass carp (Ctenopharyngodon idella)

HUANG Xiaoman , LI Ningjing , YANG Yan , SONG Jialing , TIAN Ye ,
*
LIN Li , QIN Zhendong *
(Guangzhou Key Laboratory of Aquatic Animal Diseases and Waterfowl Farming,
Guangdong Engineering Technology Research Center for Water Environment and Aquatic Product Safety,
College of Animal Science & Technology, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China)

Abstract: Iron is an essential trace element in vertebrates, involved in critical biological processes like oxygen transport and
energy metabolism. This study focused on Ctenopharyngodon idella, investigating the damaging effects of dietary iron over-
load. Healthy C. idella weighing 50-80 g were fed a diet supplemented with 0, 400, and 800 mg/kg of ferrous fumarate for 60
days. Growth performance results indicated that excessive iron intake significantly reduced the fish's length growth rate. Iron
ion content analysis showed that ferrous fumarate supplementation significantly increased iron levels in serum and liver. ELISA
results indicated that dietary iron overload led to an increase in hematocrit, along with significantly elevated activities of ALT,
AST, and GPX4 in serum and liver. qRT-PCR results revealed that iron metabolism-related genes Tf, DMT1, Hepcidin, FPN,
and NCOA4 were significantly upregulated in the ferrous fumarate group, while TfR1 and Ferritin were significantly downreg-
ulated. Furthermore, antioxidant-related genes ho-1, gpx4, and nrf2 were significantly upregulated in the ferrous fumarate
group. In vitro experiments showed that FAC incubation of L8824 cells resulted in a significant increase in intracellular iron
and ROS; qRT-PCR analysis revealed significant increases in inflammatory genes p65, tnf-α, nf-κb, tlr4, and p38-mapk, along
with significant rises in oxidative stress-related genes keap1a, nrf2, and ho-1. In conclusion, this study suggests that dietary iron
overload leads to an imbalance in iron metabolism and oxidative stress in C. idella, accompanied by inflammatory responses,
providing a theoretical basis for understanding iron metabolism-related diseases and their prevention in C. idella.
Key words: Ctenopharyngodon idella; iron overload; iron metabolism; oxidative stress; inflammatory response
Corresponding authors: LIN Li. E-mail: linli@zhku.edu.cn;
QIN Zhendong. E-mail: qinzhendongsc@163.com
Funding projects: Guangdong Basic and Applied Basic Research Foundation Natural Science Fund Project (2023A1515012784)

https://www.china-fishery.cn 中国水产学会主办 sponsored by China Society of Fisheries
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