Loss of neuronal CD200 contributed to microglial activation after acute cerebral ischemia in mice.

CD200 has been proved to play a role in immuno-inflammatory reaction. However, little information is available on CD200 in the acute stage of cerebral ischemia. We investigated the association between neuronal death and expression of CD200, and explored the relationship between CD200 and microglia in cerebral ischemic mice. Firstly, localization of CD200 expression in the normal brain tissue was detected by immunofluorescent assay. Then, focal cerebral ischemia was induced in mice by permanent middle cerebral artery occlusion (pMCAO) and then cortical tissues were collected at 6, 12, 24 and 48 h after surgery. Changes of CD200 and neuron-specific enolase (NSE) after pMCAO were assessed by western blotting. Meanwhile flow cytometry analysis was implemented to analyze the death of cortical cells. Results of these two parts were analyzed by Pearson correlation analysis. To further study, intracerebroventricular (ICV) injection of recombinant CD200 (rCD200) protein was carried out immediately after pMCAO. Iba-1 was measured by western blotting to evaluate activation of microglia, and inflammatory cytokines including IL-1β, TNF-α and IL-10 were tested by enzyme-linked immuno sorbent assay (ELISA). The results showed that CD200 was expressed in neurons and was not observed on mircroglia in cortex of normal mice. Expression of CD200 was decreased within 48 h after pMCAO, with a concomitant decrease of NSE expression. The rate of neuronal cell death was approximately around 30% and statistical analysis revealed a negative correlation between level of CD200 and the rate of neuronal death. Compared with control, exogenous rCD200 reduced expressions of Iba-1, IL-1β, TNF-α and IL-10. Taking together, our results demonstrated that loss of CD200 was caused by neuronal death and was one of contributing factors in microglial activation after cerebral ischemia. ICV injection of rCD200 protein could suppress activation of microglia in vivo.