[Chinese medicine Gandouling attenuates brain injury in hepatolenticular degeneration mice by inhibiting ferroptosis via the SIRT1/FoxO3 signaling pathway].

OBJECTIVES

To investigate the mechanism by which the Chinese medicine Gandouling protects against brain injury in hepatolenticular degeneration (Wilson disease) through regulation of the silence information regulator 1 (SIRT1)/forkhead box protein O3 (FoxO3) pathway-mediated ferroptosis.

METHODS

TX mice were randomly divided into six groups: model control, Gandouling, resveratrol (SIRT1 activator), Gandouling+resveratrol, EX-527 (SIRT1 inhibitor), and Gandouling+EX-527 groups, with DL mice serving as the blank control group. After four weeks of intervention, neurological function was assessed using the Morris water maze test, wire hanging test, and pole test. Apoptosis in basal ganglia tissue was detected by TUNEL assay. Reactive oxygen species (ROS) levels in basal ganglia tissue were measured using the DCFH-DA method, while ferrous iron (Fe²+) and malondialdehyde (MDA) levels were determined by colorimetric assays. Immunofluorescence was used to evaluate the fluorescence intensity of SIRT1 and FoxO3. The protein expression levels of SIRT1, FoxO3, glutathione peroxidase 4 (GPX4), solute carrier superfamily 7 member 11 (SLC7A11), acid-CoA synthetase long-chain family 4 (ACSL4), ferritin heavy chain 1 (FTH1), and P53 were assessed by Western blotting, and the mRNA expression levels of Sirt1 and Foxo3 were quantified by qRT-PCR.

RESULTS

Compared with the blank control group, the model control group exhibited significant neurological impairments (prolonged escape latency, reduced platform crossing frequency, decreased grip score and prolonged turning and pole-climbing time), increased neuronal apoptosis, decreased transcription and expression of Sirt1 (both P<0.01), increased transcription (P<0.05) and expression (P<0.01) of Foxo3, and elevated levels of ROS, Fe²⁺, and MDA in basal ganglia tissue (all P<0.01). Expression levels of ferroptosis-inhibiting proteins (GPX4, SLC7A11, FTH1) were decreased (all P<0.01), while those of ferroptosis-promoting proteins (P53, ACSL4) were increased (both P<0.05). Both Gandouling and resveratrol monotherapy significantly reversed the above alterations with comparable efficacy between the two treatments (all P>0.05) and combined Gandouling and resveratrol treatment exhibited synergistic effects. EX-527 exacerbated neurological impairments,neuronal apoptosis and ferroptosis (all P<0.05). However, no significant differences were observed between the Gandouling+EX-527 group and the model control group (all P>0.05).

CONCLUSIONS

Gandouling inhibits neuronal ferroptosis by regulating the SIRT1/FoxO3 signaling pathway, thereby improving neurological function in model mice.