Research Paper Volume 12, Issue 24 pp 26121—26139

Figure 5. Sub-anesthetic ISO post-conditioning inhibits OGD-led NF-κB p65 activation and PGE₂ and NO production in microglial cells in co-cultures. (A, B) At the end of 3 h-OGD or Ctrl treatment, co-cultures were exposed to RA with or without 0.7% ISO for 30 min. All the cells were continuously cultured under normal conditions for 6 or 12 h after OGD stimulation. Then, microglial cells were harvested for subsequent studies. (A) Representative western blots show total and phosphorylated IKKβ and NF-κB p65 levels at 6 h after OGD exposure. β-actin and lamin B were used as the internal controls. (B) NF-κB p65 DNA-binding activity was quantified using the TransAM NF-κB p65 transcription factor assay kit at 12 h after OGD insult. (C, D) Co-cultures with or without NAI (2 μM) pretreatment for 30 min were subjected to 3 h-OGD or Ctrl treatment and continuously cultured under normal conditions for 24 h after OGD stimulation. Then, microglial cells were collected for further analyses. (C) Quantification of PGE₂ by RIA. (D) Quantification of NO production by Griess reagent. 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 or OGD group. Ctrl: control; ISO: isoflurane; OGD: oxygen and glucose deprivation; RA: room air.