RESUMO
Ischemic stroke is a leading cause of death among human in the world, and a critical cause for long-term disability. Accumulating studies have indicated that inflammatory response regulated by microglia contributes a lot to neuronal death, but the molecular mechanism still remains unclear. V-set and immunoglobulin domain-containing 4 (Vsig4), a complement receptor of the immunoglobulin superfamily (CRIg) that specifically expresses in resting tissue-resident macrophages, plays a critical role in regulating various inflammatory diseases via multiple signaling pathways. However, the effects of Vsig4 on ischemic stroke have not been investigated. In this study, we identified that Vsig4 expression was decreased after cerebral ischemic injury induced by middle cerebral artery occlusion (MCAO). Immunofluorescence staining showed that Vsig4 was co-localized with Iba1 in microglial cells from the infarct region of MCAO-operated mice. After over-expressing Vsig4 in mice, MCAO-induced infarction area and neurological deficits score were markedly attenuated. In addition, neurological dysfunction due to MCAO surgery was improved by Vsig4 over-expression. Microglial M1 polarization was detected in mice with MCAO surgery, which was markedly inhibited by Vsig4 over-expression, as evidenced by the markedly reduced expression of CD16, CD11b, inducible nitric oxide synthase (iNOS) and interleukin 6 (IL-6); however, the expression of M2-like phenotype hallmarks such as arginase 1 (Arg1), CD206, IL-10 and Ym-1 was significantly up-regulated. Mechanistically, the anti-inflammatory role of Vsig4 was mainly through the blockage of toll-like receptor 4/nuclear factor kappa B (TLR4/NF-κB) signaling via the in vivo and in vitro experiments. Also, we found that microglial TLR4 expression in the cerebral infarct area of MCAO mice was highly suppressed by Vsig4 over-expression. In vitro, the neuron-glial mixed culture by fluorescent staining showed that oxygen glucose deprivation (OGD) treatment led to significant cell death, while being attenuated by Vsig4 over-expression in primary microglial cells. Finally, we showed that Vsig4 could interact with TLR4 and repress its expression, subsequently alleviating ischemic stroke. Collectively, our findings demonstrated that microglial Vsig4 protected against post-stroke neuro-inflammation mainly through interacting with TLR4.