Research Paper Volume 12, Issue 24 pp 26121—26139

Subanesthetic isoflurane abates ROS-activated MAPK/NF-κB signaling to repress ischemia-induced microglia inflammation and brain injury

Figure 10. Sub-anesthetic ISO post-conditioning attenuates MCAO-induced inflammation and apoptosis in rat brains. Rats were subjected to a right MCAO or sham operation (control) for 2 h and then treated with or without 0.7% ISO for 1 h. At 24 h after MCAO, seventy-six rats (Ctrl + RA, n = 20; Ctrl + ISO, n = 20; MCAO + RA, n = 17; MCAO + ISO, n = 19) still survived and then they were sacrificed under anesthesia. (A) Representative images of the TTC-stained brain tissue sections showing infarction areas (n = 8 per group). (B) Quantitative measurements of the infraction volume (n = 8 per group). (C) Histogram plots show neurologic deficit scores (n = 12 per group). (D) Quantification of rat serum PGE2 levels by RIA (n = 16 per group). (E) Quantification of rat serum NO levels by Griess reagent (n = 16 per group). (F) ELISA analysis of the rat serum IL-6 levels (n = 8 per group). (G) Analysis of ROS level by DCFH-DA assay in brain homogenates (n = 9 per group). The data represent the relative DCF fluorescence. (H) Representative western blots show total and phospho-p38 MAPK in the brain homogenates (n = 9 per group). β-actin was used as the normal control. (I) Estimation of NF-κB p65 DNA-binding activity in brain homogenates using a TransAM NF-κB p65 transcription factor assay (n = 9 per group). (J) Estimation of nucleosomal fragmentation in brain tissues (n = 9 per group). (K) Quantitative measurement of caspase-3 activity (n = 9 per group). Representative data are from three independent experiments and expressed as mean ± SD. Statistical significance: *P < 0.05 vs. Ctrl groups; #P < 0.05 vs. OGD + RA group. Ctrl: control; ISO: isoflurane; MCAO: middle cerebral arterial occlusion; RA: room air.