Fig. 8

Activation of miR-211-5p ameliorated ferroptosis and oxidative stress in hippocampal neurons of mice with KA-induced epilepsy. A Relative miR-211-5p expression in the KA, miR-211-5p agomir + KA, NC agomir + KA, miR-211-5p antagomir + KA and NC antagomir + KA groups compared with the control group. B–D The average seizure-stage score was 4.83 ± 0.41 in the KA group vs. 2.83 ± 0.75 in the miR-211-5p agomir + KA group vs. 4.67 ± 0.52 in the NC agomir + KA group vs. 4.83 ± 0.41 in the miR-211-5p antagomir + KA group vs. 4.67 ± 0.52 in the NC antagomir + KA group (B). The number of seizures within 120 min (C, average of 6 in the non-miR-211-5p agomir + KA group vs. 3 in the miR-211-5p agomir + KA group) and the average epileptiform duration of the first seizure (D, 7.08 ± 0.67 in the KA group vs. 3.87 ± 0.42 in the miR-211-5p agomir + KA group vs. 7.35 ± 0.51 in the NC agomir + KA group vs. 7.50 ± 0.61 in the miR-211-5p antagomir + KA group vs. 7.23 ± 0.49 in the NC antagomir + KA group). E Representative 5-min EEG recordings from the hippocampus (after 14 days of KA injection) showed spontaneous electrographic seizures in the KA, the miR-211-5p agomir + KA and the miR-211-5p antagomir + KA group. F The number of SRS recorded after 14 days of KA injection, demonstrating that the miR-211-5p agomir + KA group had a lesser number of seizures than those of the KA and the miR-211-5p antagomir + KA group. G The mean SRSs duration of the miR-211-5p agomir + KA group was significantly shorter than that of the KA and the miR-211-5p antagomir + KA group. H–L Western blot analysis and quantification of the protein levels of P2RX7 (I), GPX4 (J), HO-1 (K) and P-ERK/ERK (L) in the control, KA-induced epilepsy, miR-211-5p agomir + KA, NC agomir + KA, miR-211-5p antagomir + KA and NC antagomir + KA groups. M–O Representative quantification of the relative mRNA levels of P2RX7 (M), GPX4 (N) and HO-1 (O) in the control, KA, miR-211-5p agomir + KA, NC agomir + KA, miR-211-5p antagomir + KA and NC antagomir + KA groups. P–R Measurement of MDA, SOD and GSH levels to evaluate lipid peroxidation in hippocampal tissues from the abovementioned six groups. S Representative TEM images of hippocampal areas for evaluation of mitochondrial changes in randomly selected tissues. Scale bar = 200 μm or 100 μm. T Hypothesized mechanisms by which the miR-211-5p/P2RX7/ERK/GPX4 axis regulates epilepsy-associated neuronal ferroptosis and oxidative stress. All data are expressed as the mean ± SD. *p < 0.05, **p < 0.01