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 8. Sub-anesthetic ISO post-conditioning hampers ROS- or NO-mediated OGD-exposed microglial cell apoptosis in co-cultures. Co-cultures with or without NAC (5 mM) or 1400 W (50 μM) pretreatment for 30 min were stimulated with OGD or Ctrl for 3 h and subsequently exposed to RA with or without 0.7% ISO for 30 min. All the cells were continuously cultured under normal conditions for 24 h after OGD stimulation. Then, microglial cells were collected for further analyses. (A) Flow cytometry analysis of microglial cell apoptosis. (B) The nucleosomal fragmentation assay for assessing microglia apoptosis. (C) Quantitative analysis of caspase-3 activity. (D) Flow cytometry analysis of JC-1 stained microglial cells. 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; OGD: oxygen and glucose deprivation; RA: room air; NAC: N-acetyl-L-cysteine.