Subanesthetic Isoflurane Reduces Zymosan-Induced Inflammation in Murine Kupffer Cells by Inhibiting ROS-Activated p38 MAPK/NF-κB Signaling
0.7% ISO reduces ZY-induced inflammation in vivo and improves liver injury. BALB/C () mice were treated according to Section 2.3. At 8, 12, 16, or 24 h after injection with ZY or NS (Ctrl), the mice were sacrificed by euthanasia, and theirhepatic KCs and liver perfusates were isolated. ((a)–(h)) The perfusate levels of PGE2 (a) and TNF-α (b) were determined by RIA, and those of IL-1β (c), IL-6 (d), HMGB-1 (e), MIP-1α (f), MIP-2 (g), and MCP-1 (h) were determined by ELISA. (i) Fluorescence microscopy was used to determine the nuclear translocation of NF-κB p65 in the KCs immunostained with anti-NF-κB p65 antibody. (j) NF-κB DNA-binding activity of KCs was assayed by determining the optical density. (k) DCFH-DA was used to determine the generation of ROS in the KCs. (l) At 24 h after mice were injected with ZY or Ctrl, the liver tissues from each group were harvested and their morphology was assessed by hematoxylin and eosin staining. Scale bar: 5 μm. At 24 h after mice were administered with ZY or Ctrl, blood samples were collected from all groups and centrifuged to separate plasma. ((m)–(p)) The plasma levels of AST (m), ALT (n), bilirubin (o), and ALP (p) were measured by a veterinary clinical laboratory using standard laboratory techniques. Representative data are from three independent experiments and expressed as mean ± SD. versus Ctrl + RA or Ctrl + ISO; versus ZY + RA. “Time” in the figure represents the time periods of Ctrl or ZY treatment. ZY: zymosan; ISO: isoflurane; Ctrl: control; RA: room air.