Early Toll-like receptor 4 inhibition improves immune dysfunction in the hippocampus after hypoxic-ischemic brain damage

Int J Med Sci. 2022 Jan 1;19(1):142-151. doi: 10.7150/ijms.66494. eCollection 2022.


Background: Toll-like receptor 4 (TLR4) is implicated in neonatal hypoxic-ischemic brain damage (HIBD), but the underlying mechanism is unclear. Hypothesis: We hypothesized that TLR4 mediates brain damage after hypoxic ischemia (HI) by inducing abnormal neuroimmune responses, including activation of immune cells and expression disorder of immune factors, while early inhibition of TLR4 can alleviate the neuroimmune dysfunction. Method: Postnatal day 7 rats were randomized into control, HI, and HI+TAK-242 (TAK-242) groups. The HIBD model was developed using the Rice-Vannucci method (the left side was the ipsilateral side of HI). TAK-242 (0.5 mg/kg) was given to rat pups in the TAK-242 group at 30 min before modeling. Immunofluorescence, immunohistochemistry, and western blotting were used to determine the TLR4 expression; the number of Iba-1+, GFAP+, CD161+, MPO+, and CD3+ cells; ICAM-1 and C3a expression; and interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and IL-10 expression in the hippocampal CA1 region. Result: Significantly increased TLR4 expression was observed in the left hippocampus, and was alleviated by TAK-242. The significant increases in Iba-1+, MPO+, and CD161+ cells at 24 h and 7 days after HI and in GFAP+ and CD3+ T cells at 7 days after HI were also counteracted by TAK-242, but no significant differences were observed among groups at 24 h after HI. ICAM-1 expression increased 24 h after HI, while C3a expression decreased; TAK-242 also alleviated these changes. TNF-α and IL-1β expression increased, while IL-10 expression decreased at 24 h and 7 days after HI; TAK-242 counteracted the increased TNF-α and IL-1β expression at 24 h and the changes in IL-1β and IL-10 at 7 days, but induced no significant differences in IL-10 expression at 24 h and TNF-α expression at 7 days. Conclusion: Early TLR4 inhibition can alleviate hippocampal immune dysfunction after neonatal HIBD.

Keywords: Brain damage; Hippocampus; Hypoxic-ischemic; Neuroimmune; Rats; Toll-like receptor 4.

MeSH terms

  • Animals
  • Animals, Newborn
  • CA1 Region, Hippocampal / immunology
  • CA1 Region, Hippocampal / metabolism
  • CD3 Complex
  • Cytokines / metabolism
  • Female
  • Hippocampus / immunology*
  • Hippocampus / metabolism
  • Hypoxia-Ischemia, Brain / immunology*
  • Hypoxia-Ischemia, Brain / metabolism
  • Intercellular Adhesion Molecule-1 / metabolism
  • Killer Cells, Natural / metabolism
  • Male
  • Models, Animal
  • NK Cell Lectin-Like Receptor Subfamily B
  • Neutrophils / enzymology
  • Peroxidase
  • Random Allocation
  • Rats
  • Sulfonamides / pharmacology
  • T-Lymphocytes / metabolism
  • Toll-Like Receptor 4 / antagonists & inhibitors
  • Toll-Like Receptor 4 / physiology*


  • CD3 Complex
  • Cytokines
  • NK Cell Lectin-Like Receptor Subfamily B
  • Sulfonamides
  • Tlr4 protein, rat
  • Toll-Like Receptor 4
  • ethyl 6-(N-(2-chloro-4-fluorophenyl)sulfamoyl)cyclohex-1-ene-1-carboxylate
  • Intercellular Adhesion Molecule-1
  • Peroxidase