Figure 8.

LDH analysis demonstrating neuroprotective effect of V₁ agonist-induced suppression of IL-1β and TNF-α against LPS-treated ACM. Cortical neurons grown in 24-well plates were treated with ACM collected from LPS and V₁ agonist-treated cortical astrocytes followed by LDH release analysis to determine the plasma integrity of neurons. Results are expressed as mean ± SEM percentage of control (CTRL) LDH release and are representative of three separate experiments. LPS-treated ACM induced significantly higher LDH release than control, which was reversed by medium collected from LPS+V₁ agonist-treated astrocytes. Addition of IL-1β (125 pg/ml) or TNF-α (50 pg/ml) alone to neuronal cultures, together with LPS+V₁ agonist-treated ACM, had no effect on V₁ agonist-induced neuroprotection, whereas the combination of IL-1β and TNF-α reversed it. Neurons did not recover from IL-1β+TNF-α-induced cytotoxicity after cytokine removal. n = 12 per condition; ^*p < 0.05 compared with control cultures.