6-Gingerol ameliorates myocardial injury and lipid accumulation in diabetic mice: Associating with modulating ACSL3/ACSL4 expression and AMPK/Nrf2/SLC7A11 pathway activation to inhibit ferroptosis

Background To investigate the protective effects and mechanisms of 6-gingerol(6-G) on myocardial injury in diabetic mice. Methods In vivo , 18-week-old db/db mice were treated with 6-G by gavage once daily for 8 weeks. Myocardial pathological changes were assessed by H&E, WGA, Masson, Sirius red and ORO staining. In vitro , HL-1 cells were treated with high glucose and high fatty acid (HGHF) for 24 hours with or without 6-G administration. UID RNA-seq was employed to explore the protective mechanism of 6-G. ROS, MDA, GSH and SOD were examined. The expression of fatty acid transporters and ferroptosis-related proteins and genes was detected by qRT-PCR and WB. Results Myocardial injury and lipid accumulation were mitigated by 6-G. The expression of fatty acid transporters including CD36, FATP1 and FABP3 was downregulated by 6-G. Similarly, the intracellular lipid levels and the expression of CD36 and FABP3 in HL-1 cells treated with HGHF were attenuated by 6-G. UID RNA-seq analysis reveal that the protective mechanism of 6-G is associated with the AMPK pathway and ferroptosis. After 6-G treatment, the levels of ROS and MDA decreased, whereas the levels of SOD and GSH in the hearts of db/db mice. 6-G enhanced AMPK activation and upregulated the expression of Nrf2, SLC7A11 and GPX4 both in vivo and in vitro . Both ACSL3 underexpression and overexpressed ACSL4 overexpression in the model group were reversed by 6-G treatment. The protective effect and upregulation of Nrf2, SLC7A11 and GPX4 were abolished by compound C. Conclusion 6-G mitigates myocardial injury and inhibits ferroptosis via ACSL3/ACSL4 regulation and AMPK/Nrf2/SLC7A11 pathway activation.