Cytokine IL-10, activators of PI3-kinase, agonists of α-2 adrenoreceptor and antioxidants prevent ischemia-induced cell death in rat hippocampal cultures

Abstract

In the present work we compared the protective effect of anti-inflammatory cytokine IL-10 with the action of a PI3-kinase selective activator 740 Y-P, selective agonists of alpha-2 adrenoreceptor, guanfacine and UK-14,304, and compounds having antioxidant effect: recombinant human peroxiredoxin 6 and B27, in hippocampal cell culture during OGD (ischemia-like conditions). It has been shown that the response of cells to OGD in the control includes two phases. The first phase was accompanied by an increase in the frequency of spontaneous synchronous Ca²⁺-oscillations (SSCO) in neurons and Ca²⁺-pulse in astrocytes. Spontaneous Ca²⁺ events in astrocytes during ischemia in control experiments disappeared. The second phase started after a few minutes of OGD and looked like a sharp/avalanche, global synchronic (within 20 s) increase in [Ca²⁺]_i in many cells. Within 1 h after OGD, a mass death of cells, primarily astrocytes, was observed. To study the protective action of the compounds, cells were incubated in the presence of the neuroprotective agents for 10-40 min or 24 h before ischemia. All the neuroprotective agents delayed a global [Ca²⁺]_i increase during OGD or completely inhibited this process and increased cell survival.

Keywords: Alpha-2 adrenoreceptor; Antioxidant enzyme; Ca(2+) imaging; Cell survival; Fura-2; Global [Ca(2+)](i) increase; Hippocampal cell culture; Interleukin-10; Ischemia; Neuroprotective agents; OGD; PI3-Kinase.