Effect Of Fasudil On The Phenotype Conversion Of LPS-stimulated BV-2Microglia

AIM To explore the anti-inflammatory effect of Fasudil on LPS-stimulated BV-2microglia.METHODS BV-2microglia were divided into PBS control group, LPS-stimulated group and LPS plus Fasudil group. The production of NO was measured by Griess reaction. The levels of TNF-α, IL-1β, TNF-a and IL-10were detected by ELISA. The concentration of iNOS and Arg-1was determined by Western blot. The fluorescence intensity of iNOS and Arg-1was analyzed by flow cytometry. The effect of Fasudil-conditioned medium on PC-12neurons was detected by Cell Imaging Analyzer.RESULTS The treatment of LPS induced the characteristics of Ml phenotype. Fasudil inhibited the production of NO and the release of inflammatory TNF-a, IL-1β and IL-6in LPS-stimulated BV-2microglia, and promoted the release of anti-inflammatory IL-10. Fasudil decreased the content of iNOS, but increased the content of Arg-1. By flow cytometry, the mean fluorescence intensity of iNOS was decreased, while the mean fluorescence intensity of Arg-1was increased. Meanwhile, we observed the changes of nuclear density and nuclear morphology, which showed that Fasudil-conditioned medium significantly decreased PC12neuronal deathCONCLUSION Fasudil inhibits the release of inflammatory cytokines and oxidative stress reaction, resulting in an anti-inflammatory effect, which may be associated with the conversion of inflammatory Ml microglia to anti-inflammatory M2microglia. This study provides the experimental basis for the potential therapy in clinical practice.