Zinc Potentiates Lipopolysaccharide-induced Nitric Oxide Production in Cultured Primary Rat Astrocytes

Zn²⁺ plays a crucial role in the CNS where it accumulates in synaptic vesicles and is released during neurotransmission. Synaptically released Zn²⁺ is taken up by neurons and astrocytes. The majority of previous work has focused on neuronal damage caused by excess Zn²⁺. However, its effect on astrocyte function is not well understood. We examined the effect of extracellularly applied Zn²⁺ on nitric oxide (NO) production in primary cultured rat astrocytes, which were experimentally activated by lipopolysaccharide (LPS). Zn²⁺, at a concentration up to 125 μM, augmented LPS-induced NO production without affecting cell viability. LPS induced expression of both mRNA and protein of inducible NO synthase; this expression was enhanced by 125 µM Zn²⁺. Zn²⁺ also increased LPS-induced production of intracellular reactive oxygen species. Zn²⁺ enhanced the phosphorylation of p38-mitogen-activated protein kinase (MAPK) at 1–6 h after LPS treatment. The LPS-induced nuclear factor-kappaB (NFκB) activation was sustained for 6 h by Zn²⁺. Intracellular Zn²⁺ chelation with N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) or inhibition of p38-MAPK diminished the Zn²⁺ enhancement of LPS-induced NO production. These findings suggest that activation of MAPK and NFκB is important for mediating Zn²⁺enhancement of LPS-induced NO production in astrocytes. Such changes may exacerbate glial and neuronal damage during neuroinflammation.