Fig. 2

Prolonged anesthesia inducing neuroinflammation, upregulating NF-κB inflammatory pathway, downregulating neuronal excitability, and inactivating apoptotic signaling. A, B TNF-α, IL-1β, and IL-6 evidently increased in the cortex (A) and hippocampus (B) after prolonged anesthesia (n = 4 or 5 per group). C Effects of prolonged anesthesia on the morphological changes of neurons in the hippocampus. Scale bar = 20 μm. D Prolonged anesthesia activated NF-κB inflammatory pathway (n = 3 per group). E The number of Phospho-NF-κB P65-colocalized nuclei in the hippocampus (n = 4 per group). Scale bar = 10 μm. F Prolonged anesthesia inhibiting neuronal excitability marker C-fos expression in the hippocampal CA1 region (n = 4 per group). Scale bar = 20 μm. G Exhibiting representative EEG raw traces (upper) and power spectrograms (bottom) for the hippocampus. H Prolonged anesthesia triggered burst suppression in the hippocampus (n = 6 per group). The burst suppression ratio (BSR) was calculated as the percentage of suppression time of each 1 min binary series. I TUNEL staining in brain slices was negative after prolonged anesthesia. Scale bar = 100 μm. J The number of Nissl’s body (n = 4 per group). Scale bar = 20 μm. K Prolonged anesthesia had no effect on apoptotic pathways (n = 3 per group). L, M Quantification of Cleaved caspase-3/caspase-3 (L) and bcl-2/bax (M) levels normalized to β-actin. Data was shown as Mean ± SD, with *P < 0.05 or *P < 0.001; Sevo group vs. control group. Arrows represent positive cells or colocalization