Figure 4. Fer-1 improves PTE-associated cognitive deficits in mice.
(A) Experiment procedure of NOR test; (B) Statistical analysis of the exploration time, which means that the time when the mice explored the familiar or novel object, is recorded in the NOR test. N represents the novel object, and F represents the familiar object; (C) Statistical analysis of discrimination index, which was calculated using the following formula: (N-F)/(N+F) ×100%. The higher percentage of discrimination index indicates improvement of the ability to discriminate the novel object; (D) Illustration of effects of Fer-1 on swimming speed from different groups; (E) Indication of swimming traces from day 29 to day 33 after FeCl3 injection in different groups; (F-G) Indication of effects of Fer-1 on escape latency and mean path length, respectively in FeCl3-induced PTE; (H-I) Indicate representative traces in the probe trial on the day 34 after FeCl3 injection in different groups and analysis of the number of times of crossing platform. Data were expressed as mean ± SEM (n=6), *p<0.05 versus control group, **p<0.01 versus control group and #p<0.05 versus FeCl3+Vehicle group.
Figure 5. Fer-l does not display evident side effects on mice.(A) Time course of effects of Fer-1 on the body weigh from day 1 to day 34 after FeCl3 injection; (B) Cumulative analysis of effects of Fer-1 on body weight. Data were expressed as mean ± SEM (n=6).
Figure 6. Fer-l inhibits ferroptosis process in a mouse model of PTE. (A) Illustration of 4-HNE immunostaining in hippocampal subregions including CA1, CA3 and DG in different groups on the day 3 or day 34 after FeCl3 injection followed by Fer-1 treatment, Scale bar: 100 μm; (B-C) Quantitative analysis of optical intensity of 4-HNE in hippocampal CA3 regions on the day 3 and day 34 among different groups; (D-E) Measurements of effects of Fer-1 on GPx activity in FeCl3-induced PTE. Data were expressed as mean ± SEM (n=3) **p<0.01 versus control group, ***p<0.001 versus control and ##p<0.01 versus FeCl3+Vehicle group.
Figure 7. Fer-1 attenuates seizure-induced hippocampal damage in FeCl3-induced PTE. (A) Illustration of Nissl staining in hippocampal subregions including CA1, CA3 and DG in different groups on the day 3 after FeCl3 injection followed by Fer-1 treatment; (B-D) Statistical analysis of viable neurons in CA1, CA3 and DG regions on the day 3 among different groups; (E) Representative Nissl staining images in CA1, CA3 and DG regions on the day 34 after FeCl3injection followed by Fer-1 treatment; (F-H) Statistical analysis of viable neurons in CA1, CA3 and DG regions on the day 34 among different groups. Arrow head indicates dead neurons. Scale bar: 100 μm
Figure 8. Working model. It demonstrated that Fer-1 ameliorates seizures and improves cognitive impairments in FeCl3-induced PTE. Inhibition of ferroptosis process including decreases of 4-HNE level and GPx activity were possibly involved in Fer-1’s neuroprotection.