Research Paper Volume 12, Issue 24 pp 26121—26139

Figure 6. Sub-anesthetic ISO post-conditioning hinders OGD-caused activation of p38 MAPK and NF-κB p65 in microglial cells in co-cultures. (A) 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 h after OGD exposure. Then, microglial cells were collected for further analyses. Representative western blots show protein expressions of p-p38 MAPK (Thr180/Tyr182), p38 MAPK, p-JNK1/2 (Thr183/Tyr185), JNK1/2, p-ERK1/2 (Thr185/Tyr187), and ERK1/2. β-actin was used as the internal control. (B–D) Co-cultures with or without SB202190 (10 μM) pretreatment for 30 min were subjected to 3 h-OGD or Ctrl treatment and continuously cultured under normal conditions for 6 or 24 h after OGD stimulation. Then, microglial cells were collected for further analyses. (B) Representative western blots show total and phosphorylated IKKβ levels at 6 h after OGD exposure. β-actin was used as the internal control. (C) Quantification of PGE₂ levels by RIA at 24 h after OGD treatment. (D) Quantification of NO production by Griess reagent at 24 h after OGD challenge. 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.