Promotion of Analgecine on Polarization of M1 Microglia to M2 and Inhibition on Secretion of Inflammatory Factors in Vitro / 中国康复理论与实践

ABSTRACT

Objective:To investigate the effect of analgecine (AGC) on inflammatory response in the cell model of ischemic stroke and its mechanism. Methods:Sodium hydrosulfite (Na2S2O2) combined with sugar-free culture-medium was used to stimulate the model of ischemic stroke in vitro. BV2 cells were divided into six groups control group, control with 0.5 U/ml AGC group, oxygen deprivation and recovery (OGD/R) group, OGD/R with AGC (0.25 U/ml, 0.5 U/ml, 1 U/ml) groups. After oxygen and glucose deprivation for 1.5 hours, they were changed to normal medium and given different concentrations of AGC in OGD/R with AGC groups. After co-incubation for three hours, the cells were treated. The content of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the supernatant was detected. The expression of M1-type microglia marker CD16+CD32 and M2-type microglia marker CD206 were detected with immunofluorescent staining. BV2 cells were divided into seven groups control group, control with 0.5 U/ml AGC group, IL-4 group, IL-4 + lipopolysaccharide (LPS) + interferon (IFN)-γ group, IL-4 + LPS + IFN-γ with AGC (0.25 U/ml, 0.5 U/ml, 1 U/ml) groups. After 24 hours of IL-4 treatment, LPS + IFN-γ were added for 18 hours, they were changed to normal medium and given different concentrations of AGC for 24 hours, the expression of CD16+CD32 and CD206 were observed by flow cytometry. Results:Compared with the control group, the IL-6 and TNF-α level increased (P < 0.01), the number of CD16++CD32+ increased and the number of CD206+ decreased in OGD/R group. Compared with the OGD/R group, the IL-6 and TNF-α level decreased (P < 0.01), the number of CD16++CD32+ decreased and the number of CD206+ increased in AGC groups. Compared with the control group, the number of CD206 tended to increase, and the number of CD16+CD32 tended to decrease in IL-4 group; compared with IL-4 group, the number of CD16+CD32 tended to increase, and the number of CD206 tended to decrease in IL-4 + LPS + IFN-γ group; compared with IL-4 + LPS + IFN-γ group, the number of CD16+CD32 tended to decrease, and the number of CD206 tended to increase in IL-4 + LPS + IFN-γ + 0.25 U/ml AGC group and IL-4 + LPS + IFN-γ + 0.5 U/ml AGC group, while the number of CD206 increased in IL-4 + LPS + IFN-γ + 1.0 U/ml AGC group (P < 0.05). Conclusion:AGC could inhibit the secretion of inflammation factors by promoting the polarization of microglia from M1 phenotype to M2 phenotype.