Fig. 5
From: NLRP3 inflammasome inhibits mitophagy during the progression of temporal lobe epilepsy
NLRP3 inflammasome activation inhibits mitophagy and modulates seizure activity in pilocarpine-induced epileptic rats. (a) The hippocampus and temporal lobe cortex of rats were double immunofluorescence stained for TOMM20-labeled mitochondria (green) and LC3B-labeled autophagosomes (red). Nuclei were stained with DAPI (blue). Enlarged views of areas within the white boxes are shown on the right. Scale bar: 50 μm. (b) Fluorescence intensity of LC3B in the hippocampus and temporal lobe cortex of rats (n = 3). (c) Colocalization fluorescence intensity of TOMM20 and LC3B in the hippocampus and temporal lobe cortex of rats (n = 3). (d) Representative transmission electron microscopy image of the temporal cortex and hippocampus showing varying degrees of mitochondrial damage (yellow arrows) in the rat. Green arrows indicate the occurrence of mitophagy in the epileptic (EP) group. Quantitative analysis revealed that damaged mitochondria in the hippocampus of the EP group were significantly higher than in the control group. (e) Number of seizures per month in EP group and treatment groups (n = 3). The inhibitors MCC950, anti-IL -1β were applied. (f) Western blot analysis of Gasdermin D-NT, BNIP3, TOMM20, and COX IV in the hippocampal mitochondria of the control group, EP group, and treatment group. (g) Relative RNA level of the mitophagy-inhibiting molecule PPTC7 in the control group, EP group, and treatment group (n = 3). Protein levels were quantified using Image J software. Unless specified otherwise, the data are presented as means ± SEM (error bar) and compared using the two-sided Student’s t test; *P < 0.05; **P < 0.01; and ***P < 0.001; ns, no significance.
